LaserSharp 2000 Error

classic Classic list List threaded Threaded
13 messages Options
Matiar Jafari Matiar Jafari
Reply | Threaded
Open this post in threaded view
|

LaserSharp 2000 Error

I've been getting this error when i shoot larger z stacks and when i
shoot longer time series

Serious Error: NO file could be found matching the specification
'C:\Temp\untitled\untitled1_raw*.pic'

then i either click ok or close the dialog box and lasersharp 2000
closes/crashes

but when i shoot smaller stacks this does NOT happen any help would be
greatly appreciated

ive tried deleting the C:\Temp folder and this does nothing

The reason i need this to finish is so that i can then export it to my
desired location on a different server.

Thank You
---
Matiar Jafari
Michael Bastiani Michael Bastiani
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

Try increasing the memory allocated to image acquisition.  In Tools  
select System Config and set memory to size smaller than Computer  
RAM.  Also make sure your computer hard drive has plenty of space.
You cannot collect images/stacks greater than the number set in System  
Config.
good luck,
michael


On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:

> I've been getting this error when i shoot larger z stacks and when i
> shoot longer time series
>
> Serious Error: NO file could be found matching the specification
> 'C:\Temp\untitled\untitled1_raw*.pic'
>
> then i either click ok or close the dialog box and lasersharp 2000
> closes/crashes
>
> but when i shoot smaller stacks this does NOT happen any help would be
> greatly appreciated
>
> ive tried deleting the C:\Temp folder and this does nothing
>
> The reason i need this to finish is so that i can then export it to my
> desired location on a different server.
>
> Thank You
> ---
> Matiar Jafari

Michael Bastiani
[hidden email]
W. Chan W. Chan
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

Hi Matiar

It may be that the drive where your temp directory is located is running
short of space thus preventing acquistion.  LS2K writes files to c:\temp
first, in your case, as they are acquired as untitledN ... and them copy
(or move?) to the directory you specified.  In general, acquistion does
not depend on the amount of RAM installed.

As Michael suggested, go to Tools | System configuration, look under the
Scan System tab for Collection Limits to make sure that it is set
properly.  Depending on the verion of LS2K, it could be either 1 GB (pre
5.2, if I remember correctly) or 2 GB.  This limit is hardcoded and cannot
be increased.  If your stack will go over that limit LS2K will not let
you.  You can also change where the Temporary Directory is located, may be
point it to a larger disk.

You do need to have system access level in LS2K to change things in System
config, I think.  You can check that in Tools | User Setup.  Good luck.

On Tue, 24 Mar 2009, Michael Bastiani wrote:

> Try increasing the memory allocated to image acquisition.  In Tools select
> System Config and set memory to size smaller than Computer RAM.  Also make
> sure your computer hard drive has plenty of space.
> You cannot collect images/stacks greater than the number set in System Config.
> good luck,
> michael
>
>
> On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
>
> > I've been getting this error when i shoot larger z stacks and when i
> > shoot longer time series
> >
> > Serious Error: NO file could be found matching the specification
> > 'C:\Temp\untitled\untitled1_raw*.pic'
> >
> > then i either click ok or close the dialog box and lasersharp 2000
> > closes/crashes
> >
> > but when i shoot smaller stacks this does NOT happen any help would be
> > greatly appreciated
> >
> > ive tried deleting the C:\Temp folder and this does nothing
> >
> > The reason i need this to finish is so that i can then export it to my
> > desired location on a different server.
> >
> > Thank You
> > ---
> > Matiar Jafari

--
Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
Keith Morris Keith Morris
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

In reply to this post by Michael Bastiani
Hi Matiar,
Chang and others have beaten me too it but anyway:

I've had this error before on our Radiance 2000 and it was fixed pretty
easily and permanently after a chat to the Bio-Rad engineer, but it was
sorted out years ago so I am rather fuzzy on the reason for it and the cure.
I'm sure though it is a memory error of some sort [I assume you are still
running LaserSharp 2000 under NT - our error wasn't on the latest version 6
but earlier, probably v4]. We did add more physical memory - to 768Mb or
possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
matched pairs], plus we played around with the hard drive space, system page
file, LaserSharp settings logged in as system, and we put the crucial :\temp
on our very much larger D:\ drive [set via Lasersharp system] as the C:\
drive is always compromised to ~4Gb under NT, unless you increased it after
SP6 is applied. We had two hard drives fitted, the first split into two
partitions C:,E: and the much larger second one D: for user data and the
Temp folder  

So I'm sure all the other postings are on the right track, it's system or
hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what error
message we got though other than the Z-stacks/time-lapses never finished and
we lost everything]. I think we still had to split very long time-lapses
with z stacks though into two runs [2Gb limit]. Also allocated more virtual
memory [page file] from the hard drive to help out the physical RAM
[forgotten how to do that under NT, but it's probably under 'system']. I
think there used to be a 2Gb limit on collections though even with
LaserSharp 6 [1Gb default] even with the tweeks.

Zeiss [Microscience] should still support the Radiance 2000 [not the 1024
though] so an email to their service support should clarify the solution. I
don't know if upgrading to XP ever helped [you needed a new Firewire card
and deep pockets for Bio-Rad to do this for you].

Again look at the old postings below that seem very relevant.

Keith

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford  OX3 7BN,
United Kingdom.

Telephone:  +44 (0)1865 287568
Email:  [hidden email]
Web-pages: http://www.well.ox.ac.uk/cytogenetics/







-------------------------------------------------------------------
Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
From: W. Chan
Reply-To: Confocal Microscopy List <[hidden email]>
Date: Tue, 21 Mar 2006 16:59:39 -0800
Content-Type: TEXT/PLAIN

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

The Control Panel error message is quite misleading.  For v5.2, it states
that "Collection Requires Too Much Memory" when the predicted dataset size
exceeds the collection limit, but in fact it refers to disk space.

I agree.  Even a 2 GB limit is really not much when you need
multi-channels with a merge pane at 1024x1024 with multiple time points.
May be a work around is not to have a merge pane which can be generated
quite easily after you have the data.  A few of our users do not use
the merge pane option in their methods.  They do not use LaserSharp to
analyze their images so they can't read the merge pane .pmf file even if
they want to.

--
Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)

On Sat, 18 Mar 2006, Michael Cammer wrote:

> I'll check the upgrade software for writing to disk.  But with multiple
> probe files our problem was the merge file.  For two probes, it needed
> 150% more memory than the raw images themselves.  Even changing the
> configuration didn't provide enough memory.
>
>
>> Are you referring to RAM or harddisk space?  As far as I know,
LaserSharps
>> writes to temporary files rather than holding the experiment in RAM.  We
>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with LaserSharp
>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
>> "System Configuration | Scan system" for the setting of the collection
>> limit.  Previous versions may have a 1 GB limit.  Also check the location
>> of the temporary files which needs to have plenty of space (as much as
you

>> want to acquire each time or more).  Good luck.
>>
>> --
>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>>
>> On Fri, 17 Mar 2006, Michael Cammer wrote:
>>
>>> We have the Radiance 2000.  This is a major limitation of the system.
>>> We've had to compromise by restarting long time lapse experiments in the
>>> middle, by taking fewer Z planes per timepoint or longer intervals.
>>> But when our BioRad confocals die, LaserSharp software should be
>>> obsolete.
>>> -Michael
>>>
>>> At 05:51 PM 03/14/06 -0800, you wrote:
>>>>
>>>> I have a question regarding a BioRad 1024 confocal system.  This system
>>>> is
>>>> working fine.  But one trouble is that people cannot run any scanning
>>>> program (time lapse experiments) that contains more than 230 mb,
>>>> although
>>>> the system has plenty of free memory (we just added another 1gb to it).
>>>>  I
>>>> will appreciate if anyone can share with us if you had same experience
>>>> and
>>>> managed to resolve it.








RECENT POSTINGS
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Michael Bastiani
Sent: 24 March 2009 13:57
To: [hidden email]
Subject: Re: LaserSharp 2000 Error

Try increasing the memory allocated to image acquisition.  In Tools  
select System Config and set memory to size smaller than Computer  
RAM.  Also make sure your computer hard drive has plenty of space.
You cannot collect images/stacks greater than the number set in System  
Config.
good luck,
michael


On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:

> I've been getting this error when i shoot larger z stacks and when i
> shoot longer time series
>
> Serious Error: NO file could be found matching the specification
> 'C:\Temp\untitled\untitled1_raw*.pic'
>
> then i either click ok or close the dialog box and lasersharp 2000
> closes/crashes
>
> but when i shoot smaller stacks this does NOT happen any help would be
> greatly appreciated
>
> ive tried deleting the C:\Temp folder and this does nothing
>
> The reason i need this to finish is so that i can then export it to my
> desired location on a different server.
>
> Thank You
> ---
> Matiar Jafari

Michael Bastiani
[hidden email]
Matiar Jafari Matiar Jafari
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

Dear all,

So according to Chan the limit is hardcoded and cannot be changed....

we have multiple accounts on the computer and im wondering if im not
able to change he memory option from 700 mb to the 1gb because its
hardcoded or because i need the "admin" login user for lasersharp
2000....help? also when i change the directory for the temp files it
doesn't allow me to click ok thus it doesnt change where its
located....any help???

On Tue, Mar 24, 2009 at 11:12 AM, Keith Morris <[hidden email]> wrote:

> Hi Matiar,
> Chang and others have beaten me too it but anyway:
>
> I've had this error before on our Radiance 2000 and it was fixed pretty
> easily and permanently after a chat to the Bio-Rad engineer, but it was
> sorted out years ago so I am rather fuzzy on the reason for it and the cure.
> I'm sure though it is a memory error of some sort [I assume you are still
> running LaserSharp 2000 under NT - our error wasn't on the latest version 6
> but earlier, probably v4]. We did add more physical memory - to 768Mb or
> possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
> matched pairs], plus we played around with the hard drive space, system page
> file, LaserSharp settings logged in as system, and we put the crucial :\temp
> on our very much larger D:\ drive [set via Lasersharp system] as the C:\
> drive is always compromised to ~4Gb under NT, unless you increased it after
> SP6 is applied. We had two hard drives fitted, the first split into two
> partitions C:,E: and the much larger second one D: for user data and the
> Temp folder
>
> So I'm sure all the other postings are on the right track, it's system or
> hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what error
> message we got though other than the Z-stacks/time-lapses never finished and
> we lost everything]. I think we still had to split very long time-lapses
> with z stacks though into two runs [2Gb limit]. Also allocated more virtual
> memory [page file] from the hard drive to help out the physical RAM
> [forgotten how to do that under NT, but it's probably under 'system']. I
> think there used to be a 2Gb limit on collections though even with
> LaserSharp 6 [1Gb default] even with the tweeks.
>
> Zeiss [Microscience] should still support the Radiance 2000 [not the 1024
> though] so an email to their service support should clarify the solution. I
> don't know if upgrading to XP ever helped [you needed a new Firewire card
> and deep pockets for Bio-Rad to do this for you].
>
> Again look at the old postings below that seem very relevant.
>
> Keith
>
> ---------------------------------------------------------------------------
> Dr Keith J. Morris,
> Molecular Cytogenetics and Microscopy Core,
> Laboratory 00/069 and 00/070,
> The Wellcome Trust Centre for Human Genetics,
> Roosevelt Drive,
> Oxford  OX3 7BN,
> United Kingdom.
>
> Telephone:  +44 (0)1865 287568
> Email:  [hidden email]
> Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>
>
>
>
>
>
>
> -------------------------------------------------------------------
> Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
> From: W. Chan
> Reply-To: Confocal Microscopy List <[hidden email]>
> Date: Tue, 21 Mar 2006 16:59:39 -0800
> Content-Type: TEXT/PLAIN
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> The Control Panel error message is quite misleading.  For v5.2, it states
> that "Collection Requires Too Much Memory" when the predicted dataset size
> exceeds the collection limit, but in fact it refers to disk space.
>
> I agree.  Even a 2 GB limit is really not much when you need
> multi-channels with a merge pane at 1024x1024 with multiple time points.
> May be a work around is not to have a merge pane which can be generated
> quite easily after you have the data.  A few of our users do not use
> the merge pane option in their methods.  They do not use LaserSharp to
> analyze their images so they can't read the merge pane .pmf file even if
> they want to.
>
> --
> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>
> On Sat, 18 Mar 2006, Michael Cammer wrote:
>
>> I'll check the upgrade software for writing to disk.  But with multiple
>> probe files our problem was the merge file.  For two probes, it needed
>> 150% more memory than the raw images themselves.  Even changing the
>> configuration didn't provide enough memory.
>>
>>
>>> Are you referring to RAM or harddisk space?  As far as I know,
> LaserSharps
>>> writes to temporary files rather than holding the experiment in RAM.  We
>>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with LaserSharp
>>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
>>> "System Configuration | Scan system" for the setting of the collection
>>> limit.  Previous versions may have a 1 GB limit.  Also check the location
>>> of the temporary files which needs to have plenty of space (as much as
> you
>>> want to acquire each time or more).  Good luck.
>>>
>>> --
>>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>>>
>>> On Fri, 17 Mar 2006, Michael Cammer wrote:
>>>
>>>> We have the Radiance 2000.  This is a major limitation of the system.
>>>> We've had to compromise by restarting long time lapse experiments in the
>>>> middle, by taking fewer Z planes per timepoint or longer intervals.
>>>> But when our BioRad confocals die, LaserSharp software should be
>>>> obsolete.
>>>> -Michael
>>>>
>>>> At 05:51 PM 03/14/06 -0800, you wrote:
>>>>>
>>>>> I have a question regarding a BioRad 1024 confocal system.  This system
>>>>> is
>>>>> working fine.  But one trouble is that people cannot run any scanning
>>>>> program (time lapse experiments) that contains more than 230 mb,
>>>>> although
>>>>> the system has plenty of free memory (we just added another 1gb to it).
>>>>>  I
>>>>> will appreciate if anyone can share with us if you had same experience
>>>>> and
>>>>> managed to resolve it.
>
>
>
>
>
>
>
>
> RECENT POSTINGS
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On
> Behalf Of Michael Bastiani
> Sent: 24 March 2009 13:57
> To: [hidden email]
> Subject: Re: LaserSharp 2000 Error
>
> Try increasing the memory allocated to image acquisition.  In Tools
> select System Config and set memory to size smaller than Computer
> RAM.  Also make sure your computer hard drive has plenty of space.
> You cannot collect images/stacks greater than the number set in System
> Config.
> good luck,
> michael
>
>
> On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
>
>> I've been getting this error when i shoot larger z stacks and when i
>> shoot longer time series
>>
>> Serious Error: NO file could be found matching the specification
>> 'C:\Temp\untitled\untitled1_raw*.pic'
>>
>> then i either click ok or close the dialog box and lasersharp 2000
>> closes/crashes
>>
>> but when i shoot smaller stacks this does NOT happen any help would be
>> greatly appreciated
>>
>> ive tried deleting the C:\Temp folder and this does nothing
>>
>> The reason i need this to finish is so that i can then export it to my
>> desired location on a different server.
>>
>> Thank You
>> ---
>> Matiar Jafari
>
> Michael Bastiani
> [hidden email]
>



--
Matiar Jafari
W. Chan W. Chan
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

Hi Matiar

What I meant was e.g., in pre-5.2, you can only give max 1 GB to the
application.  It is not RAM related.  Since you did not mention what
version and OS you're running, I just assumed you're still using NT.
LaserSharp 6 and XP run really well together; but that's another story.  I
only have access to v6 now so the terms in the menu may be different than
earlier version.

System access in lasersharp is completely independent of NT's (or whatever
window's) user privilege.  Usually, the software is set up with an user
called Administrator that have system access, other users for the software
may only have read & write access.  After you log into lasersharp, go to
Tools | User Setup, the second column will tell you what Access level you
have.  Whoever install the software should have the password to log in as
Administrator.

There is a way to find that out.  Everything for this software are stored
in a database e.g., c:\LaserSharpNT\Database\lsMessages.mdb.  If you have
microsoft access, you can open a COPY of this file and look at the table
called LaserSharpUsers and you can see all the user passwords in plain
text.  Access level 0 is for system.  It is probably overly cautious to
open a copy because the database will likely be opened as read-only; but
why take a chance.

Once you log in as an user with system access level, you should be able to
change all the configuration parameters.  But be very careful, you can
mess up things really bad.  I've learned that the hard way.  Hope this
help.

On Tue, 24 Mar 2009, Matiar Jafari wrote:

> Dear all,
>
> So according to Chan the limit is hardcoded and cannot be changed....
>
> we have multiple accounts on the computer and im wondering if im not
> able to change he memory option from 700 mb to the 1gb because its
> hardcoded or because i need the "admin" login user for lasersharp
> 2000....help? also when i change the directory for the temp files it
> doesn't allow me to click ok thus it doesnt change where its
> located....any help???
>
> On Tue, Mar 24, 2009 at 11:12 AM, Keith Morris <[hidden email]> wrote:
> > Hi Matiar,
> > Chang and others have beaten me too it but anyway:
> >
> > I've had this error before on our Radiance 2000 and it was fixed pretty
> > easily and permanently after a chat to the Bio-Rad engineer, but it was
> > sorted out years ago so I am rather fuzzy on the reason for it and the cure.
> > I'm sure though it is a memory error of some sort [I assume you are still
> > running LaserSharp 2000 under NT - our error wasn't on the latest version 6
> > but earlier, probably v4]. We did add more physical memory - to 768Mb or
> > possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
> > matched pairs], plus we played around with the hard drive space, system page
> > file, LaserSharp settings logged in as system, and we put the crucial :\temp
> > on our very much larger D:\ drive [set via Lasersharp system] as the C:\
> > drive is always compromised to ~4Gb under NT, unless you increased it after
> > SP6 is applied. We had two hard drives fitted, the first split into two
> > partitions C:,E: and the much larger second one D: for user data and the
> > Temp folder
> >
> > So I'm sure all the other postings are on the right track, it's system or
> > hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what error
> > message we got though other than the Z-stacks/time-lapses never finished and
> > we lost everything]. I think we still had to split very long time-lapses
> > with z stacks though into two runs [2Gb limit]. Also allocated more virtual
> > memory [page file] from the hard drive to help out the physical RAM
> > [forgotten how to do that under NT, but it's probably under 'system']. I
> > think there used to be a 2Gb limit on collections though even with
> > LaserSharp 6 [1Gb default] even with the tweeks.
> >
> > Zeiss [Microscience] should still support the Radiance 2000 [not the 1024
> > though] so an email to their service support should clarify the solution. I
> > don't know if upgrading to XP ever helped [you needed a new Firewire card
> > and deep pockets for Bio-Rad to do this for you].
> >
> > Again look at the old postings below that seem very relevant.
> >
> > Keith
> >
> > ---------------------------------------------------------------------------
> > Dr Keith J. Morris,
> > Molecular Cytogenetics and Microscopy Core,
> > Laboratory 00/069 and 00/070,
> > The Wellcome Trust Centre for Human Genetics,
> > Roosevelt Drive,
> > Oxford  OX3 7BN,
> > United Kingdom.
> >
> > Telephone:  +44 (0)1865 287568
> > Email:  [hidden email]
> > Web-pages: http://www.well.ox.ac.uk/cytogenetics/
> >
> >
> >
> >
> >
> >
> >
> > -------------------------------------------------------------------
> > Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
> > From: W. Chan
> > Reply-To: Confocal Microscopy List <[hidden email]>
> > Date: Tue, 21 Mar 2006 16:59:39 -0800
> > Content-Type: TEXT/PLAIN
> >
> > Search the CONFOCAL archive at
> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> > The Control Panel error message is quite misleading.  For v5.2, it states
> > that "Collection Requires Too Much Memory" when the predicted dataset size
> > exceeds the collection limit, but in fact it refers to disk space.
> >
> > I agree.  Even a 2 GB limit is really not much when you need
> > multi-channels with a merge pane at 1024x1024 with multiple time points.
> > May be a work around is not to have a merge pane which can be generated
> > quite easily after you have the data.  A few of our users do not use
> > the merge pane option in their methods.  They do not use LaserSharp to
> > analyze their images so they can't read the merge pane .pmf file even if
> > they want to.
> >
> > --
> > Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
> > The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
> >
> > On Sat, 18 Mar 2006, Michael Cammer wrote:
> >
> >> I'll check the upgrade software for writing to disk.  But with multiple
> >> probe files our problem was the merge file.  For two probes, it needed
> >> 150% more memory than the raw images themselves.  Even changing the
> >> configuration didn't provide enough memory.
> >>
> >>
> >>> Are you referring to RAM or harddisk space?  As far as I know,
> > LaserSharps
> >>> writes to temporary files rather than holding the experiment in RAM.  We
> >>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with LaserSharp
> >>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
> >>> "System Configuration | Scan system" for the setting of the collection
> >>> limit.  Previous versions may have a 1 GB limit.  Also check the location
> >>> of the temporary files which needs to have plenty of space (as much as
> > you
> >>> want to acquire each time or more).  Good luck.
> >>>
> >>> --
> >>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
> >>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
> >>>
> >>> On Fri, 17 Mar 2006, Michael Cammer wrote:
> >>>
> >>>> We have the Radiance 2000.  This is a major limitation of the system.
> >>>> We've had to compromise by restarting long time lapse experiments in the
> >>>> middle, by taking fewer Z planes per timepoint or longer intervals.
> >>>> But when our BioRad confocals die, LaserSharp software should be
> >>>> obsolete.
> >>>> -Michael
> >>>>
> >>>> At 05:51 PM 03/14/06 -0800, you wrote:
> >>>>>
> >>>>> I have a question regarding a BioRad 1024 confocal system.  This system
> >>>>> is
> >>>>> working fine.  But one trouble is that people cannot run any scanning
> >>>>> program (time lapse experiments) that contains more than 230 mb,
> >>>>> although
> >>>>> the system has plenty of free memory (we just added another 1gb to it).
> >>>>>  I
> >>>>> will appreciate if anyone can share with us if you had same experience
> >>>>> and
> >>>>> managed to resolve it.
> >
> > RECENT POSTINGS
> > -----Original Message-----
> > From: Confocal Microscopy List [mailto:[hidden email]] On
> > Behalf Of Michael Bastiani
> > Sent: 24 March 2009 13:57
> > To: [hidden email]
> > Subject: Re: LaserSharp 2000 Error
> >
> > Try increasing the memory allocated to image acquisition.  In Tools
> > select System Config and set memory to size smaller than Computer
> > RAM.  Also make sure your computer hard drive has plenty of space.
> > You cannot collect images/stacks greater than the number set in System
> > Config.
> > good luck,
> > michael
> >
> >
> > On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
> >
> >> I've been getting this error when i shoot larger z stacks and when i
> >> shoot longer time series
> >>
> >> Serious Error: NO file could be found matching the specification
> >> 'C:\Temp\untitled\untitled1_raw*.pic'
> >>
> >> then i either click ok or close the dialog box and lasersharp 2000
> >> closes/crashes
> >>
> >> but when i shoot smaller stacks this does NOT happen any help would be
> >> greatly appreciated
> >>
> >> ive tried deleting the C:\Temp folder and this does nothing
> >>
> >> The reason i need this to finish is so that i can then export it to my
> >> desired location on a different server.
> >>
> >> Thank You
> >> ---
> >> Matiar Jafari
--
Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
Prof Hindsight Prof Hindsight
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

Dear Matiar Jafari,
 
Have you actually contacted a Carl Zeiss service representative about this problem?
 
From my experience and that of others, Zeiss will assist with such problems.  I also have it on good authority that although one can no longer buy a service/maintenance contract for an MRC-1024, they will endeavour to service the system on a billable basis for on-site work and, as Bio-Rad did in the past, offer remote assistance at no cost.
Whether you use Zeiss for support or not, knowing a bit more about what type of confocal you actually have and its configuration would be helpful and avoid misdiagnosis.
 
Good luck,
 
PH
On Wed, Mar 25, 2009 at 1:24 AM, W. Chan <[hidden email]> wrote:
Hi Matiar

What I meant was e.g., in pre-5.2, you can only give max 1 GB to the
application.  It is not RAM related.  Since you did not mention what
version and OS you're running, I just assumed you're still using NT.
LaserSharp 6 and XP run really well together; but that's another story.  I
only have access to v6 now so the terms in the menu may be different than
earlier version.

System access in lasersharp is completely independent of NT's (or whatever
window's) user privilege.  Usually, the software is set up with an user
called Administrator that have system access, other users for the software
may only have read & write access.  After you log into lasersharp, go to
Tools | User Setup, the second column will tell you what Access level you
have.  Whoever install the software should have the password to log in as
Administrator.

There is a way to find that out.  Everything for this software are stored
in a database e.g., c:\LaserSharpNT\Database\lsMessages.mdb.  If you have
microsoft access, you can open a COPY of this file and look at the table
called LaserSharpUsers and you can see all the user passwords in plain
text.  Access level 0 is for system.  It is probably overly cautious to
open a copy because the database will likely be opened as read-only; but
why take a chance.

Once you log in as an user with system access level, you should be able to
change all the configuration parameters.  But be very careful, you can
mess up things really bad.  I've learned that the hard way.  Hope this
help.

On Tue, 24 Mar 2009, Matiar Jafari wrote:

> Dear all,
>
> So according to Chan the limit is hardcoded and cannot be changed....
>
> we have multiple accounts on the computer and im wondering if im not
> able to change he memory option from 700 mb to the 1gb because its
> hardcoded or because i need the "admin" login user for lasersharp
> 2000....help? also when i change the directory for the temp files it
> doesn't allow me to click ok thus it doesnt change where its
> located....any help???
>
> On Tue, Mar 24, 2009 at 11:12 AM, Keith Morris <[hidden email]> wrote:
> > Hi Matiar,
> > Chang and others have beaten me too it but anyway:
> >
> > I've had this error before on our Radiance 2000 and it was fixed pretty
> > easily and permanently after a chat to the Bio-Rad engineer, but it was
> > sorted out years ago so I am rather fuzzy on the reason for it and the cure.
> > I'm sure though it is a memory error of some sort [I assume you are still
> > running LaserSharp 2000 under NT - our error wasn't on the latest version 6
> > but earlier, probably v4]. We did add more physical memory - to 768Mb or
> > possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
> > matched pairs], plus we played around with the hard drive space, system page
> > file, LaserSharp settings logged in as system, and we put the crucial :\temp
> > on our very much larger D:\ drive [set via Lasersharp system] as the C:\
> > drive is always compromised to ~4Gb under NT, unless you increased it after
> > SP6 is applied. We had two hard drives fitted, the first split into two
> > partitions C:,E: and the much larger second one D: for user data and the
> > Temp folder
> >
> > So I'm sure all the other postings are on the right track, it's system or
> > hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what error
> > message we got though other than the Z-stacks/time-lapses never finished and
> > we lost everything]. I think we still had to split very long time-lapses
> > with z stacks though into two runs [2Gb limit]. Also allocated more virtual
> > memory [page file] from the hard drive to help out the physical RAM
> > [forgotten how to do that under NT, but it's probably under 'system']. I
> > think there used to be a 2Gb limit on collections though even with
> > LaserSharp 6 [1Gb default] even with the tweeks.
> >
> > Zeiss [Microscience] should still support the Radiance 2000 [not the 1024
> > though] so an email to their service support should clarify the solution. I
> > don't know if upgrading to XP ever helped [you needed a new Firewire card
> > and deep pockets for Bio-Rad to do this for you].
> >
> > Again look at the old postings below that seem very relevant.
> >
> > Keith
> >
> > ---------------------------------------------------------------------------
> > Dr Keith J. Morris,
> > Molecular Cytogenetics and Microscopy Core,
> > Laboratory 00/069 and 00/070,
> > The Wellcome Trust Centre for Human Genetics,
> > Roosevelt Drive,
> > Oxford  OX3 7BN,
> > United Kingdom.
> >
> > Telephone:  +44 (0)1865 287568
> > Email:  [hidden email]
> > Web-pages: http://www.well.ox.ac.uk/cytogenetics/
> >
> >
> >
> >
> >
> >
> >
> > -------------------------------------------------------------------
> > Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
> > From: W. Chan
> > Reply-To: Confocal Microscopy List <[hidden email]>
> > Date: Tue, 21 Mar 2006 16:59:39 -0800
> > Content-Type: TEXT/PLAIN
> >
> > Search the CONFOCAL archive at
> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> > The Control Panel error message is quite misleading.  For v5.2, it states
> > that "Collection Requires Too Much Memory" when the predicted dataset size
> > exceeds the collection limit, but in fact it refers to disk space.
> >
> > I agree.  Even a 2 GB limit is really not much when you need
> > multi-channels with a merge pane at 1024x1024 with multiple time points.
> > May be a work around is not to have a merge pane which can be generated
> > quite easily after you have the data.  A few of our users do not use
> > the merge pane option in their methods.  They do not use LaserSharp to
> > analyze their images so they can't read the merge pane .pmf file even if
> > they want to.
> >
> > --
> > Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
> > The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
> >
> > On Sat, 18 Mar 2006, Michael Cammer wrote:
> >
> >> I'll check the upgrade software for writing to disk.  But with multiple
> >> probe files our problem was the merge file.  For two probes, it needed
> >> 150% more memory than the raw images themselves.  Even changing the
> >> configuration didn't provide enough memory.
> >>
> >>
> >>> Are you referring to RAM or harddisk space?  As far as I know,
> > LaserSharps
> >>> writes to temporary files rather than holding the experiment in RAM.  We
> >>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with LaserSharp
> >>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
> >>> "System Configuration | Scan system" for the setting of the collection
> >>> limit.  Previous versions may have a 1 GB limit.  Also check the location
> >>> of the temporary files which needs to have plenty of space (as much as
> > you
> >>> want to acquire each time or more).  Good luck.
> >>>
> >>> --
> >>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
> >>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
> >>>
> >>> On Fri, 17 Mar 2006, Michael Cammer wrote:
> >>>
> >>>> We have the Radiance 2000.  This is a major limitation of the system.
> >>>> We've had to compromise by restarting long time lapse experiments in the
> >>>> middle, by taking fewer Z planes per timepoint or longer intervals.
> >>>> But when our BioRad confocals die, LaserSharp software should be
> >>>> obsolete.
> >>>> -Michael
> >>>>
> >>>> At 05:51 PM 03/14/06 -0800, you wrote:
> >>>>>
> >>>>> I have a question regarding a BioRad 1024 confocal system.  This system
> >>>>> is
> >>>>> working fine.  But one trouble is that people cannot run any scanning
> >>>>> program (time lapse experiments) that contains more than 230 mb,
> >>>>> although
> >>>>> the system has plenty of free memory (we just added another 1gb to it).
> >>>>>  I
> >>>>> will appreciate if anyone can share with us if you had same experience
> >>>>> and
> >>>>> managed to resolve it.
> >
> > RECENT POSTINGS
> > -----Original Message-----
> > From: Confocal Microscopy List [mailto:[hidden email]] On
> > Behalf Of Michael Bastiani
> > Sent: 24 March 2009 13:57
> > To: [hidden email]
> > Subject: Re: LaserSharp 2000 Error
> >
> > Try increasing the memory allocated to image acquisition.  In Tools
> > select System Config and set memory to size smaller than Computer
> > RAM.  Also make sure your computer hard drive has plenty of space.
> > You cannot collect images/stacks greater than the number set in System
> > Config.
> > good luck,
> > michael
> >
> >
> > On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
> >
> >> I've been getting this error when i shoot larger z stacks and when i
> >> shoot longer time series
> >>
> >> Serious Error: NO file could be found matching the specification
> >> 'C:\Temp\untitled\untitled1_raw*.pic'
> >>
> >> then i either click ok or close the dialog box and lasersharp 2000
> >> closes/crashes
> >>
> >> but when i shoot smaller stacks this does NOT happen any help would be
> >> greatly appreciated
> >>
> >> ive tried deleting the C:\Temp folder and this does nothing
> >>
> >> The reason i need this to finish is so that i can then export it to my
> >> desired location on a different server.
> >>
> >> Thank You
> >> ---
> >> Matiar Jafari

--
Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)

Matiar Jafari Matiar Jafari
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

New error message after the first one I get. Failed to load PicMat with data fileException in channels, stop capture

I changed the ram capacity to 2gb and I tries changing the harddrive temp.

I'm shooting then exporting but I can't export as it gives me the error after I shoot anything larger than~80um

Thank You
---
Matiar Jafari

On Mar 25, 2009, at 2:51 AM, Prof Hindsight <[hidden email]> wrote:

Dear Matiar Jafari,
 
Have you actually contacted a Carl Zeiss service representative about this problem?
 
From my experience and that of others, Zeiss will assist with such problems.  I also have it on good authority that although one can no longer buy a service/maintenance contract for an MRC-1024, they will endeavour to service the system on a billable basis for on-site work and, as Bio-Rad did in the past, offer remote assistance at no cost.
Whether you use Zeiss for support or not, knowing a bit more about what type of confocal you actually have and its configuration would be helpful and avoid misdiagnosis.
 
Good luck,
 
PH
On Wed, Mar 25, 2009 at 1:24 AM, W. Chan <[hidden email]> wrote:
Hi Matiar

What I meant was e.g., in pre-5.2, you can only give max 1 GB to the
application.  It is not RAM related.  Since you did not mention what
version and OS you're running, I just assumed you're still using NT.
LaserSharp 6 and XP run really well together; but that's another story.  I
only have access to v6 now so the terms in the menu may be different than
earlier version.

System access in lasersharp is completely independent of NT's (or whatever
window's) user privilege.  Usually, the software is set up with an user
called Administrator that have system access, other users for the software
may only have read & write access.  After you log into lasersharp, go to
Tools | User Setup, the second column will tell you what Access level you
have.  Whoever install the software should have the password to log in as
Administrator.

There is a way to find that out.  Everything for this software are stored
in a database e.g., c:\LaserSharpNT\Database\lsMessages.mdb.  If you have
microsoft access, you can open a COPY of this file and look at the table
called LaserSharpUsers and you can see all the user passwords in plain
text.  Access level 0 is for system.  It is probably overly cautious to
open a copy because the database will likely be opened as read-only; but
why take a chance.

Once you log in as an user with system access level, you should be able to
change all the configuration parameters.  But be very careful, you can
mess up things really bad.  I've learned that the hard way.  Hope this
help.

On Tue, 24 Mar 2009, Matiar Jafari wrote:

> Dear all,
>
> So according to Chan the limit is hardcoded and cannot be changed....
>
> we have multiple accounts on the computer and im wondering if im not
> able to change he memory option from 700 mb to the 1gb because its
> hardcoded or because i need the "admin" login user for lasersharp
> 2000....help? also when i change the directory for the temp files it
> doesn't allow me to click ok thus it doesnt change where its
> located....any help???
>
> On Tue, Mar 24, 2009 at 11:12 AM, Keith Morris <[hidden email]> wrote:
> > Hi Matiar,
> > Chang and others have beaten me too it but anyway:
> >
> > I've had this error before on our Radiance 2000 and it was fixed pretty
> > easily and permanently after a chat to the Bio-Rad engineer, but it was
> > sorted out years ago so I am rather fuzzy on the reason for it and the cure.
> > I'm sure though it is a memory error of some sort [I assume you are still
> > running LaserSharp 2000 under NT - our error wasn't on the latest version 6
> > but earlier, probably v4]. We did add more physical memory - to 768Mb or
> > possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
> > matched pairs], plus we played around with the hard drive space, system page
> > file, LaserSharp settings logged in as system, and we put the crucial :\temp
> > on our very much larger D:\ drive [set via Lasersharp system] as the C:\
> > drive is always compromised to ~4Gb under NT, unless you increased it after
> > SP6 is applied. We had two hard drives fitted, the first split into two
> > partitions C:,E: and the much larger second one D: for user data and the
> > Temp folder
> >
> > So I'm sure all the other postings are on the right track, it's system or
> > hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what error
> > message we got though other than the Z-stacks/time-lapses never finished and
> > we lost everything]. I think we still had to split very long time-lapses
> > with z stacks though into two runs [2Gb limit]. Also allocated more virtual
> > memory [page file] from the hard drive to help out the physical RAM
> > [forgotten how to do that under NT, but it's probably under 'system']. I
> > think there used to be a 2Gb limit on collections though even with
> > LaserSharp 6 [1Gb default] even with the tweeks.
> >
> > Zeiss [Microscience] should still support the Radiance 2000 [not the 1024
> > though] so an email to their service support should clarify the solution. I
> > don't know if upgrading to XP ever helped [you needed a new Firewire card
> > and deep pockets for Bio-Rad to do this for you].
> >
> > Again look at the old postings below that seem very relevant.
> >
> > Keith
> >
> > ---------------------------------------------------------------------------
> > Dr Keith J. Morris,
> > Molecular Cytogenetics and Microscopy Core,
> > Laboratory 00/069 and 00/070,
> > The Wellcome Trust Centre for Human Genetics,
> > Roosevelt Drive,
> > Oxford  OX3 7BN,
> > United Kingdom.
> >
> > Telephone:  +44 (0)1865 287568
> > Email:  [hidden email]
> > Web-pages: http://www.well.ox.ac.uk/cytogenetics/
> >
> >
> >
> >
> >
> >
> >
> > -------------------------------------------------------------------
> > Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
> > From: W. Chan
> > Reply-To: Confocal Microscopy List <[hidden email]>
> > Date: Tue, 21 Mar 2006 16:59:39 -0800
> > Content-Type: TEXT/PLAIN
> >
> > Search the CONFOCAL archive at
> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> > The Control Panel error message is quite misleading.  For v5.2, it states
> > that "Collection Requires Too Much Memory" when the predicted dataset size
> > exceeds the collection limit, but in fact it refers to disk space.
> >
> > I agree.  Even a 2 GB limit is really not much when you need
> > multi-channels with a merge pane at 1024x1024 with multiple time points.
> > May be a work around is not to have a merge pane which can be generated
> > quite easily after you have the data.  A few of our users do not use
> > the merge pane option in their methods.  They do not use LaserSharp to
> > analyze their images so they can't read the merge pane .pmf file even if
> > they want to.
> >
> > --
> > Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
> > The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
> >
> > On Sat, 18 Mar 2006, Michael Cammer wrote:
> >
> >> I'll check the upgrade software for writing to disk.  But with multiple
> >> probe files our problem was the merge file.  For two probes, it needed
> >> 150% more memory than the raw images themselves.  Even changing the
> >> configuration didn't provide enough memory.
> >>
> >>
> >>> Are you referring to RAM or harddisk space?  As far as I know,
> > LaserSharps
> >>> writes to temporary files rather than holding the experiment in RAM.  We
> >>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with LaserSharp
> >>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
> >>> "System Configuration | Scan system" for the setting of the collection
> >>> limit.  Previous versions may have a 1 GB limit.  Also check the location
> >>> of the temporary files which needs to have plenty of space (as much as
> > you
> >>> want to acquire each time or more).  Good luck.
> >>>
> >>> --
> >>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
> >>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
> >>>
> >>> On Fri, 17 Mar 2006, Michael Cammer wrote:
> >>>
> >>>> We have the Radiance 2000.  This is a major limitation of the system.
> >>>> We've had to compromise by restarting long time lapse experiments in the
> >>>> middle, by taking fewer Z planes per timepoint or longer intervals.
> >>>> But when our BioRad confocals die, LaserSharp software should be
> >>>> obsolete.
> >>>> -Michael
> >>>>
> >>>> At 05:51 PM 03/14/06 -0800, you wrote:
> >>>>>
> >>>>> I have a question regarding a BioRad 1024 confocal system.  This system
> >>>>> is
> >>>>> working fine.  But one trouble is that people cannot run any scanning
> >>>>> program (time lapse experiments) that contains more than 230 mb,
> >>>>> although
> >>>>> the system has plenty of free memory (we just added another 1gb to it).
> >>>>>  I
> >>>>> will appreciate if anyone can share with us if you had same experience
> >>>>> and
> >>>>> managed to resolve it.
> >
> > RECENT POSTINGS
> > -----Original Message-----
> > From: Confocal Microscopy List [mailto:[hidden email]] On
> > Behalf Of Michael Bastiani
> > Sent: 24 March 2009 13:57
> > To: [hidden email]
> > Subject: Re: LaserSharp 2000 Error
> >
> > Try increasing the memory allocated to image acquisition.  In Tools
> > select System Config and set memory to size smaller than Computer
> > RAM.  Also make sure your computer hard drive has plenty of space.
> > You cannot collect images/stacks greater than the number set in System
> > Config.
> > good luck,
> > michael
> >
> >
> > On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
> >
> >> I've been getting this error when i shoot larger z stacks and when i
> >> shoot longer time series
> >>
> >> Serious Error: NO file could be found matching the specification
> >> 'C:\Temp\untitled\untitled1_raw*.pic'
> >>
> >> then i either click ok or close the dialog box and lasersharp 2000
> >> closes/crashes
> >>
> >> but when i shoot smaller stacks this does NOT happen any help would be
> >> greatly appreciated
> >>
> >> ive tried deleting the C:\Temp folder and this does nothing
> >>
> >> The reason i need this to finish is so that i can then export it to my
> >> desired location on a different server.
> >>
> >> Thank You
> >> ---
> >> Matiar Jafari

--
Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)

Keith Morris Keith Morris
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

In reply to this post by Prof Hindsight

More memory with NT/Lasersharp only helps if you are really down at the 256Mb level, and mostly it gives stability/speed provided the modules are matched [all ECC] because you probably still have that 1Gb to 2Gb upper limit built into Lasersharp. Besides the hard drive page-file can [slowly] provide more memory. Check your PC motherboard can cope with 2Gb physical RAM [older systems have 1Gb and even 512Mb limits] – crucial.com often gives this info. Don’t know if you have a MRC1024 or Radiance, but Zeiss [Microscience] should be able to advise if your error messages are consistent with Lasersharp v4/v5 operating normally or it’s an error [still sound like its hitting the Lasersharp image capture memory limit of 1Gb or 2Gb depending on version – unless of course you used to be able to capture larger z-stacks]. Once you hit that Lasersharp limit, it’s either an upgrade or you have to break the stack into a couple of separate ones. From memory when an error message talks specifically about ‘not enough memory for capture’ it frequently means hard drive space not system memory.

 

Our Bio-Rad 1024 was no longer fully supported by Zeiss Micro-Science a few years ago now and so we passed it on to those less fortunate and bought a Zeiss 510 Meta. Zeiss did say they would continue to repair the MRC 1024 system while they could still get parts, and that they would continue to provide technical support advice [which should be free via telephone/email].

 

Many Bio-Rad staff have obviously left the company since UK Bio-Rad confocal production was stopped by Zeiss, although they retained core personal for product support of the newer Radiance 2000 confocals that also run LaserSharp - some of whom will still remember the elderly 1024 [and perhaps even COMOS, let alone LaserSharp]. Plus all the knowledgeable Bio-Rad confocal reps generally live on working for other microscope companies.

 

Keith

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford  OX3 7BN,
United Kingdom.

Telephone:  +44 (0)1865 287568
Email:  [hidden email]
Web-pages:
http://www.well.ox.ac.uk/cytogenetics/

New error message after the first one I get. Failed to load PicMat with data fileException in channels, stop capture

 

I changed the ram capacity to 2gb and I tries changing the harddrive temp.

 

I'm shooting then exporting but I can't export as it gives me the error after I shoot anything larger than~80um

Thank You

---

Matiar Jafari


On Mar 25, 2009, at 2:51 AM, Prof Hindsight <[hidden email]> wrote:

Dear Matiar Jafari,

 

Have you actually contacted a Carl Zeiss service representative about this problem?

 

From my experience and that of others, Zeiss will assist with such problems.  I also have it on good authority that although one can no longer buy a service/maintenance contract for an MRC-1024, they will endeavour to service the system on a billable basis for on-site work and, as Bio-Rad did in the past, offer remote assistance at no cost.

Whether you use Zeiss for support or not, knowing a bit more about what type of confocal you actually have and its configuration would be helpful and avoid misdiagnosis.

 

Good luck,

 

PH

On Wed, Mar 25, 2009 at 1:24 AM, W. Chan <[hidden email][hidden email]> wrote:

Hi Matiar

What I meant was e.g., in pre-5.2, you can only give max 1 GB to the
application.  It is not RAM related.  Since you did not mention what
version and OS you're running, I just assumed you're still using NT.
LaserSharp 6 and XP run really well together; but that's another story.  I
only have access to v6 now so the terms in the menu may be different than
earlier version.

System access in lasersharp is completely independent of NT's (or whatever
window's) user privilege.  Usually, the software is set up with an user
called Administrator that have system access, other users for the software
may only have read & write access.  After you log into lasersharp, go to
Tools | User Setup, the second column will tell you what Access level you
have.  Whoever install the software should have the password to log in as
Administrator.

There is a way to find that out.  Everything for this software are stored
in a database e.g., c:\LaserSharpNT\Database\lsMessages.mdb.  If you have
microsoft access, you can open a COPY of this file and look at the table
called LaserSharpUsers and you can see all the user passwords in plain
text.  Access level 0 is for system.  It is probably overly cautious to
open a copy because the database will likely be opened as read-only; but
why take a chance.

Once you log in as an user with system access level, you should be able to
change all the configuration parameters.  But be very careful, you can
mess up things really bad.  I've learned that the hard way.  Hope this
help.


On Tue, 24 Mar 2009, Matiar Jafari wrote:

> Dear all,
>
> So according to Chan the limit is hardcoded and cannot be changed....
>
> we have multiple accounts on the computer and im wondering if im not
> able to change he memory option from 700 mb to the 1gb because its
> hardcoded or because i need the "admin" login user for lasersharp
> 2000....help? also when i change the directory for the temp files it
> doesn't allow me to click ok thus it doesnt change where its
> located....any help???
>
> On Tue, Mar 24, 2009 at 11:12 AM, Keith Morris <[hidden email][hidden email]> wrote:
> > Hi Matiar,
> > Chang and others have beaten me too it but anyway:
> >
> > I've had this error before on our Radiance 2000 and it was fixed pretty
> > easily and permanently after a chat to the Bio-Rad engineer, but it was
> > sorted out years ago so I am rather fuzzy on the reason for it and the cure.
> > I'm sure though it is a memory error of some sort [I assume you are still
> > running LaserSharp 2000 under NT - our error wasn't on the latest version 6
> > but earlier, probably v4]. We did add more physical memory - to 768Mb or
> > possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
> > matched pairs], plus we played around with the hard drive space, system page
> > file, LaserSharp settings logged in as system, and we put the crucial :\temp
> > on our very much larger D:\ drive [set via Lasersharp system] as the C:\
> > drive is always compromised to ~4Gb under NT, unless you increased it after
> > SP6 is applied. We had two hard drives fitted, the first split into two
> > partitions C:,E: and the much larger second one D: for user data and the
> > Temp folder
> >
> > So I'm sure all the other postings are on the right track, it's system or
> > hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what error
> > message we got though other than the Z-stacks/time-lapses never finished and
> > we lost everything]. I think we still had to split very long time-lapses
> > with z stacks though into two runs [2Gb limit]. Also allocated more virtual
> > memory [page file] from the hard drive to help out the physical RAM
> > [forgotten how to do that under NT, but it's probably under 'system']. I
> > think there used to be a 2Gb limit on collections though even with
> > LaserSharp 6 [1Gb default] even with the tweeks.
> >
> > Zeiss [Microscience] should still support the Radiance 2000 [not the 1024
> > though] so an email to their service support should clarify the solution. I
> > don't know if upgrading to XP ever helped [you needed a new Firewire card
> > and deep pockets for Bio-Rad to do this for you].
> >
> > Again look at the old postings below that seem very relevant.
> >
> > Keith
> >
> > ---------------------------------------------------------------------------
> > Dr Keith J. Morris,
> > Molecular Cytogenetics and Microscopy Core,
> > Laboratory 00/069 and 00/070,
> > The Wellcome Trust Centre for Human Genetics,
> > Roosevelt Drive,
> > Oxford  OX3 7BN,
> > United Kingdom.
> >
> > Telephone:  +44 (0)1865 287568
> > Email:  [hidden email][hidden email]
> > Web-pages: http://www.well.ox.ac.uk/cytogenetics/
> >
> >
> >
> >
> >
> >
> >
> > -------------------------------------------------------------------
> > Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
> > From: W. Chan
> > Reply-To: Confocal Microscopy List <[hidden email][hidden email]>
> > Date: Tue, 21 Mar 2006 16:59:39 -0800
> > Content-Type: TEXT/PLAIN
> >
> > Search the CONFOCAL archive at
> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> >
> > The Control Panel error message is quite misleading.  For v5.2, it states
> > that "Collection Requires Too Much Memory" when the predicted dataset size
> > exceeds the collection limit, but in fact it refers to disk space.
> >
> > I agree.  Even a 2 GB limit is really not much when you need
> > multi-channels with a merge pane at 1024x1024 with multiple time points.
> > May be a work around is not to have a merge pane which can be generated
> > quite easily after you have the data.  A few of our users do not use
> > the merge pane option in their methods.  They do not use LaserSharp to
> > analyze their images so they can't read the merge pane .pmf file even if
> > they want to.
> >
> > --
> > Pang (Wai Pang Chan, [hidden email][hidden email], PAB A087, 206-685-1519)
> > The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
> >
> > On Sat, 18 Mar 2006, Michael Cammer wrote:
> >
> >> I'll check the upgrade software for writing to disk.  But with multiple
> >> probe files our problem was the merge file.  For two probes, it needed
> >> 150% more memory than the raw images themselves.  Even changing the
> >> configuration didn't provide enough memory.
> >>
> >>
> >>> Are you referring to RAM or harddisk space?  As far as I know,
> > LaserSharps
> >>> writes to temporary files rather than holding the experiment in RAM.  We
> >>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with LaserSharp
> >>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
> >>> "System Configuration | Scan system" for the setting of the collection
> >>> limit.  Previous versions may have a 1 GB limit.  Also check the location
> >>> of the temporary files which needs to have plenty of space (as much as
> > you
> >>> want to acquire each time or more).  Good luck.
> >>>
> >>> --
> >>> Pang (Wai Pang Chan, [hidden email][hidden email], PAB A087, 206-685-1519)
> >>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
> >>>
> >>> On Fri, 17 Mar 2006, Michael Cammer wrote:
> >>>
> >>>> We have the Radiance 2000.  This is a major limitation of the system.
> >>>> We've had to compromise by restarting long time lapse experiments in the
> >>>> middle, by taking fewer Z planes per timepoint or longer intervals.
> >>>> But when our BioRad confocals die, LaserSharp software should be
> >>>> obsolete.
> >>>> -Michael
> >>>>
> >>>> At 05:51 PM 03/14/06 -0800, you wrote:
> >>>>>
> >>>>> I have a question regarding a BioRad 1024 confocal system.  This system
> >>>>> is
> >>>>> working fine.  But one trouble is that people cannot run any scanning
> >>>>> program (time lapse experiments) that contains more than 230 mb,
> >>>>> although
> >>>>> the system has plenty of free memory (we just added another 1gb to it).
> >>>>>  I
> >>>>> will appreciate if anyone can share with us if you had same experience
> >>>>> and
> >>>>> managed to resolve it.
> >
> > RECENT POSTINGS
> > -----Original Message-----
> > From: Confocal Microscopy List [mailto:[hidden email][hidden email]] On
> > Behalf Of Michael Bastiani
> > Sent: 24 March 2009 13:57
> > To: [hidden email][hidden email]
> > Subject: Re: LaserSharp 2000 Error
> >
> > Try increasing the memory allocated to image acquisition.  In Tools
> > select System Config and set memory to size smaller than Computer
> > RAM.  Also make sure your computer hard drive has plenty of space.
> > You cannot collect images/stacks greater than the number set in System
> > Config.
> > good luck,
> > michael
> >
> >
> > On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
> >
> >> I've been getting this error when i shoot larger z stacks and when i
> >> shoot longer time series
> >>
> >> Serious Error: NO file could be found matching the specification
> >> 'C:\Temp\untitled\untitled1_raw*.pic'
> >>
> >> then i either click ok or close the dialog box and lasersharp 2000
> >> closes/crashes
> >>
> >> but when i shoot smaller stacks this does NOT happen any help would be
> >> greatly appreciated
> >>
> >> ive tried deleting the C:\Temp folder and this does nothing
> >>
> >> The reason i need this to finish is so that i can then export it to my
> >> desired location on a different server.
> >>
> >> Thank You
> >> ---
> >> Matiar Jafari

--

Pang (Wai Pang Chan, [hidden email][hidden email], PAB A087, 206-685-1519)
The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)

 

 

Matiar Jafari Matiar Jafari
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

when i try to acquire >80um stacks its at about either 80-100mb(i
forgot not sure)  The system is running on windowns NT version 6
build(i forget i could check later)  The error has always been there
since ive started working with the confocal and since others have been
using it as well.  Ive changed the hard drive temp save area from my c
drive(has something around 70gb of free space) to the D drive which
only has like 10gb of free sapce and ive tried changing it to a server
which has 200gb of free space.  Ive also played with the RAM changed
it from 1000Mb to 2000MB still no dice

I called a zeiss representative yesterday and he said that its a known
bug and there are two fixes to create a new channel with the laser off
or to install a service pack which he was supposed to email me but i
never received.

system has only 1gb of memory.  But still i dont think this matters as
i am ONLY capturing a stack that is around 80mb.  So far the farthest
ive been able to get with largest z stack is 28 um which is about 50
MB

On Fri, Mar 27, 2009 at 2:42 AM, Keith Morris <[hidden email]> wrote:

> More memory with NT/Lasersharp only helps if you are really down at the
> 256Mb level, and mostly it gives stability/speed provided the modules are
> matched [all ECC] because you probably still have that 1Gb to 2Gb upper
> limit built into Lasersharp. Besides the hard drive page-file can [slowly]
> provide more memory. Check your PC motherboard can cope with 2Gb physical
> RAM [older systems have 1Gb and even 512Mb limits] – crucial.com often gives
> this info. Don’t know if you have a MRC1024 or Radiance, but Zeiss
> [Microscience] should be able to advise if your error messages are
> consistent with Lasersharp v4/v5 operating normally or it’s an error [still
> sound like its hitting the Lasersharp image capture memory limit of 1Gb or
> 2Gb depending on version – unless of course you used to be able to capture
> larger z-stacks]. Once you hit that Lasersharp limit, it’s either an upgrade
> or you have to break the stack into a couple of separate ones. From memory
> when an error message talks specifically about ‘not enough memory for
> capture’ it frequently means hard drive space not system memory.
>
>
>
> Our Bio-Rad 1024 was no longer fully supported by Zeiss Micro-Science a few
> years ago now and so we passed it on to those less fortunate and bought a
> Zeiss 510 Meta. Zeiss did say they would continue to repair the MRC 1024
> system while they could still get parts, and that they would continue to
> provide technical support advice [which should be free via telephone/email].
>
>
>
> Many Bio-Rad staff have obviously left the company since UK Bio-Rad confocal
> production was stopped by Zeiss, although they retained core personal for
> product support of the newer Radiance 2000 confocals that also run
> LaserSharp - some of whom will still remember the elderly 1024 [and perhaps
> even COMOS, let alone LaserSharp]. Plus all the knowledgeable Bio-Rad
> confocal reps generally live on working for other microscope companies.
>
>
>
> Keith
>
> ---------------------------------------------------------------------------
> Dr Keith J. Morris,
> Molecular Cytogenetics and Microscopy Core,
> Laboratory 00/069 and 00/070,
> The Wellcome Trust Centre for Human Genetics,
> Roosevelt Drive,
> Oxford  OX3 7BN,
> United Kingdom.
>
> Telephone:  +44 (0)1865 287568
> Email:  [hidden email]
> Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>
> New error message after the first one I get. Failed to load PicMat with data
> fileException in channels, stop capture
>
>
>
> I changed the ram capacity to 2gb and I tries changing the harddrive temp.
>
>
>
> I'm shooting then exporting but I can't export as it gives me the error
> after I shoot anything larger than~80um
>
> Thank You
>
> ---
>
> Matiar Jafari
>
> On Mar 25, 2009, at 2:51 AM, Prof Hindsight <[hidden email]>
> wrote:
>
> Dear Matiar Jafari,
>
>
>
> Have you actually contacted a Carl Zeiss service representative about this
> problem?
>
>
>
> From my experience and that of others, Zeiss will assist with such
> problems.  I also have it on good authority that although one can no longer
> buy a service/maintenance contract for an MRC-1024, they will endeavour to
> service the system on a billable basis for on-site work and, as Bio-Rad did
> in the past, offer remote assistance at no cost.
>
> Whether you use Zeiss for support or not, knowing a bit more about what type
> of confocal you actually have and its configuration would be helpful and
> avoid misdiagnosis.
>
>
>
> Good luck,
>
>
>
> PH
>
> On Wed, Mar 25, 2009 at 1:24 AM, W. Chan <[hidden email]> wrote:
>
> Hi Matiar
>
> What I meant was e.g., in pre-5.2, you can only give max 1 GB to the
> application.  It is not RAM related.  Since you did not mention what
> version and OS you're running, I just assumed you're still using NT.
> LaserSharp 6 and XP run really well together; but that's another story.  I
> only have access to v6 now so the terms in the menu may be different than
> earlier version.
>
> System access in lasersharp is completely independent of NT's (or whatever
> window's) user privilege.  Usually, the software is set up with an user
> called Administrator that have system access, other users for the software
> may only have read & write access.  After you log into lasersharp, go to
> Tools | User Setup, the second column will tell you what Access level you
> have.  Whoever install the software should have the password to log in as
> Administrator.
>
> There is a way to find that out.  Everything for this software are stored
> in a database e.g., c:\LaserSharpNT\Database\lsMessages.mdb.  If you have
> microsoft access, you can open a COPY of this file and look at the table
> called LaserSharpUsers and you can see all the user passwords in plain
> text.  Access level 0 is for system.  It is probably overly cautious to
> open a copy because the database will likely be opened as read-only; but
> why take a chance.
>
> Once you log in as an user with system access level, you should be able to
> change all the configuration parameters.  But be very careful, you can
> mess up things really bad.  I've learned that the hard way.  Hope this
> help.
>
> On Tue, 24 Mar 2009, Matiar Jafari wrote:
>
>> Dear all,
>>
>> So according to Chan the limit is hardcoded and cannot be changed....
>>
>> we have multiple accounts on the computer and im wondering if im not
>> able to change he memory option from 700 mb to the 1gb because its
>> hardcoded or because i need the "admin" login user for lasersharp
>> 2000....help? also when i change the directory for the temp files it
>> doesn't allow me to click ok thus it doesnt change where its
>> located....any help???
>>
>> On Tue, Mar 24, 2009 at 11:12 AM, Keith Morris <[hidden email]>
>> wrote:
>> > Hi Matiar,
>> > Chang and others have beaten me too it but anyway:
>> >
>> > I've had this error before on our Radiance 2000 and it was fixed pretty
>> > easily and permanently after a chat to the Bio-Rad engineer, but it was
>> > sorted out years ago so I am rather fuzzy on the reason for it and the
>> > cure.
>> > I'm sure though it is a memory error of some sort [I assume you are
>> > still
>> > running LaserSharp 2000 under NT - our error wasn't on the latest
>> > version 6
>> > but earlier, probably v4]. We did add more physical memory - to 768Mb or
>> > possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
>> > matched pairs], plus we played around with the hard drive space, system
>> > page
>> > file, LaserSharp settings logged in as system, and we put the crucial
>> > :\temp
>> > on our very much larger D:\ drive [set via Lasersharp system] as the C:\
>> > drive is always compromised to ~4Gb under NT, unless you increased it
>> > after
>> > SP6 is applied. We had two hard drives fitted, the first split into two
>> > partitions C:,E: and the much larger second one D: for user data and the
>> > Temp folder
>> >
>> > So I'm sure all the other postings are on the right track, it's system
>> > or
>> > hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what
>> > error
>> > message we got though other than the Z-stacks/time-lapses never finished
>> > and
>> > we lost everything]. I think we still had to split very long time-lapses
>> > with z stacks though into two runs [2Gb limit]. Also allocated more
>> > virtual
>> > memory [page file] from the hard drive to help out the physical RAM
>> > [forgotten how to do that under NT, but it's probably under 'system']. I
>> > think there used to be a 2Gb limit on collections though even with
>> > LaserSharp 6 [1Gb default] even with the tweeks.
>> >
>> > Zeiss [Microscience] should still support the Radiance 2000 [not the
>> > 1024
>> > though] so an email to their service support should clarify the
>> > solution. I
>> > don't know if upgrading to XP ever helped [you needed a new Firewire
>> > card
>> > and deep pockets for Bio-Rad to do this for you].
>> >
>> > Again look at the old postings below that seem very relevant.
>> >
>> > Keith
>> >
>> >
>> > ---------------------------------------------------------------------------
>> > Dr Keith J. Morris,
>> > Molecular Cytogenetics and Microscopy Core,
>> > Laboratory 00/069 and 00/070,
>> > The Wellcome Trust Centre for Human Genetics,
>> > Roosevelt Drive,
>> > Oxford  OX3 7BN,
>> > United Kingdom.
>> >
>> > Telephone:  +44 (0)1865 287568
>> > Email:  [hidden email]
>> > Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>> >
>> >
>> >
>> >
>> >
>> >
>> >
>> > -------------------------------------------------------------------
>> > Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
>> > From: W. Chan
>> > Reply-To: Confocal Microscopy List <[hidden email]>
>> > Date: Tue, 21 Mar 2006 16:59:39 -0800
>> > Content-Type: TEXT/PLAIN
>> >
>> > Search the CONFOCAL archive at
>> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>> >
>> > The Control Panel error message is quite misleading.  For v5.2, it
>> > states
>> > that "Collection Requires Too Much Memory" when the predicted dataset
>> > size
>> > exceeds the collection limit, but in fact it refers to disk space.
>> >
>> > I agree.  Even a 2 GB limit is really not much when you need
>> > multi-channels with a merge pane at 1024x1024 with multiple time points.
>> > May be a work around is not to have a merge pane which can be generated
>> > quite easily after you have the data.  A few of our users do not use
>> > the merge pane option in their methods.  They do not use LaserSharp to
>> > analyze their images so they can't read the merge pane .pmf file even if
>> > they want to.
>> >
>> > --
>> > Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>> > The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>> >
>> > On Sat, 18 Mar 2006, Michael Cammer wrote:
>> >
>> >> I'll check the upgrade software for writing to disk.  But with multiple
>> >> probe files our problem was the merge file.  For two probes, it needed
>> >> 150% more memory than the raw images themselves.  Even changing the
>> >> configuration didn't provide enough memory.
>> >>
>> >>
>> >>> Are you referring to RAM or harddisk space?  As far as I know,
>> > LaserSharps
>> >>> writes to temporary files rather than holding the experiment in RAM.
>> >>>  We
>> >>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with
>> >>> LaserSharp
>> >>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
>> >>> "System Configuration | Scan system" for the setting of the collection
>> >>> limit.  Previous versions may have a 1 GB limit.  Also check the
>> >>> location
>> >>> of the temporary files which needs to have plenty of space (as much as
>> > you
>> >>> want to acquire each time or more).  Good luck.
>> >>>
>> >>> --
>> >>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>> >>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>> >>>
>> >>> On Fri, 17 Mar 2006, Michael Cammer wrote:
>> >>>
>> >>>> We have the Radiance 2000.  This is a major limitation of the system.
>> >>>> We've had to compromise by restarting long time lapse experiments in
>> >>>> the
>> >>>> middle, by taking fewer Z planes per timepoint or longer intervals.
>> >>>> But when our BioRad confocals die, LaserSharp software should be
>> >>>> obsolete.
>> >>>> -Michael
>> >>>>
>> >>>> At 05:51 PM 03/14/06 -0800, you wrote:
>> >>>>>
>> >>>>> I have a question regarding a BioRad 1024 confocal system.  This
>> >>>>> system
>> >>>>> is
>> >>>>> working fine.  But one trouble is that people cannot run any
>> >>>>> scanning
>> >>>>> program (time lapse experiments) that contains more than 230 mb,
>> >>>>> although
>> >>>>> the system has plenty of free memory (we just added another 1gb to
>> >>>>> it).
>> >>>>>  I
>> >>>>> will appreciate if anyone can share with us if you had same
>> >>>>> experience
>> >>>>> and
>> >>>>> managed to resolve it.
>> >
>> > RECENT POSTINGS
>> > -----Original Message-----
>> > From: Confocal Microscopy List [mailto:[hidden email]]
>> > On
>> > Behalf Of Michael Bastiani
>> > Sent: 24 March 2009 13:57
>> > To: [hidden email]
>> > Subject: Re: LaserSharp 2000 Error
>> >
>> > Try increasing the memory allocated to image acquisition.  In Tools
>> > select System Config and set memory to size smaller than Computer
>> > RAM.  Also make sure your computer hard drive has plenty of space.
>> > You cannot collect images/stacks greater than the number set in System
>> > Config.
>> > good luck,
>> > michael
>> >
>> >
>> > On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
>> >
>> >> I've been getting this error when i shoot larger z stacks and when i
>> >> shoot longer time series
>> >>
>> >> Serious Error: NO file could be found matching the specification
>> >> 'C:\Temp\untitled\untitled1_raw*.pic'
>> >>
>> >> then i either click ok or close the dialog box and lasersharp 2000
>> >> closes/crashes
>> >>
>> >> but when i shoot smaller stacks this does NOT happen any help would be
>> >> greatly appreciated
>> >>
>> >> ive tried deleting the C:\Temp folder and this does nothing
>> >>
>> >> The reason i need this to finish is so that i can then export it to my
>> >> desired location on a different server.
>> >>
>> >> Thank You
>> >> ---
>> >> Matiar Jafari
>
> --
>
> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>
>
>
>



--
Matiar Jafari
Matiar Jafari Matiar Jafari
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

In reply to this post by Keith Morris
update... my apologies i made a mistake im running windows 2000 with
Lasersharp 2000 6.0 build 846 i just shot a stack that was only 19 um
and it gave me the same error.

I was told the work around was to

"....the only solution is to use a method with 2 channels to collect
data.  This will allow you to collect
up to the limits set by LaserSharp ~2GB.  Once you have your data you
can delete the empty/redundant channel of data."  Zeiss Support

im shooting with a green method with only one channel any help in how
i would do this?

thanks in advance


Thank You

On Fri, Mar 27, 2009 at 2:42 AM, Keith Morris <[hidden email]> wrote:

> More memory with NT/Lasersharp only helps if you are really down at the
> 256Mb level, and mostly it gives stability/speed provided the modules are
> matched [all ECC] because you probably still have that 1Gb to 2Gb upper
> limit built into Lasersharp. Besides the hard drive page-file can [slowly]
> provide more memory. Check your PC motherboard can cope with 2Gb physical
> RAM [older systems have 1Gb and even 512Mb limits] – crucial.com often gives
> this info. Don’t know if you have a MRC1024 or Radiance, but Zeiss
> [Microscience] should be able to advise if your error messages are
> consistent with Lasersharp v4/v5 operating normally or it’s an error [still
> sound like its hitting the Lasersharp image capture memory limit of 1Gb or
> 2Gb depending on version – unless of course you used to be able to capture
> larger z-stacks]. Once you hit that Lasersharp limit, it’s either an upgrade
> or you have to break the stack into a couple of separate ones. From memory
> when an error message talks specifically about ‘not enough memory for
> capture’ it frequently means hard drive space not system memory.
>
>
>
> Our Bio-Rad 1024 was no longer fully supported by Zeiss Micro-Science a few
> years ago now and so we passed it on to those less fortunate and bought a
> Zeiss 510 Meta. Zeiss did say they would continue to repair the MRC 1024
> system while they could still get parts, and that they would continue to
> provide technical support advice [which should be free via telephone/email].
>
>
>
> Many Bio-Rad staff have obviously left the company since UK Bio-Rad confocal
> production was stopped by Zeiss, although they retained core personal for
> product support of the newer Radiance 2000 confocals that also run
> LaserSharp - some of whom will still remember the elderly 1024 [and perhaps
> even COMOS, let alone LaserSharp]. Plus all the knowledgeable Bio-Rad
> confocal reps generally live on working for other microscope companies.
>
>
>
> Keith
>
> ---------------------------------------------------------------------------
> Dr Keith J. Morris,
> Molecular Cytogenetics and Microscopy Core,
> Laboratory 00/069 and 00/070,
> The Wellcome Trust Centre for Human Genetics,
> Roosevelt Drive,
> Oxford  OX3 7BN,
> United Kingdom.
>
> Telephone:  +44 (0)1865 287568
> Email:  [hidden email]
> Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>
> New error message after the first one I get. Failed to load PicMat with data
> fileException in channels, stop capture
>
>
>
> I changed the ram capacity to 2gb and I tries changing the harddrive temp.
>
>
>
> I'm shooting then exporting but I can't export as it gives me the error
> after I shoot anything larger than~80um
>
> Thank You
>
> ---
>
> Matiar Jafari
>
> On Mar 25, 2009, at 2:51 AM, Prof Hindsight <[hidden email]>
> wrote:
>
> Dear Matiar Jafari,
>
>
>
> Have you actually contacted a Carl Zeiss service representative about this
> problem?
>
>
>
> From my experience and that of others, Zeiss will assist with such
> problems.  I also have it on good authority that although one can no longer
> buy a service/maintenance contract for an MRC-1024, they will endeavour to
> service the system on a billable basis for on-site work and, as Bio-Rad did
> in the past, offer remote assistance at no cost.
>
> Whether you use Zeiss for support or not, knowing a bit more about what type
> of confocal you actually have and its configuration would be helpful and
> avoid misdiagnosis.
>
>
>
> Good luck,
>
>
>
> PH
>
> On Wed, Mar 25, 2009 at 1:24 AM, W. Chan <[hidden email]> wrote:
>
> Hi Matiar
>
> What I meant was e.g., in pre-5.2, you can only give max 1 GB to the
> application.  It is not RAM related.  Since you did not mention what
> version and OS you're running, I just assumed you're still using NT.
> LaserSharp 6 and XP run really well together; but that's another story.  I
> only have access to v6 now so the terms in the menu may be different than
> earlier version.
>
> System access in lasersharp is completely independent of NT's (or whatever
> window's) user privilege.  Usually, the software is set up with an user
> called Administrator that have system access, other users for the software
> may only have read & write access.  After you log into lasersharp, go to
> Tools | User Setup, the second column will tell you what Access level you
> have.  Whoever install the software should have the password to log in as
> Administrator.
>
> There is a way to find that out.  Everything for this software are stored
> in a database e.g., c:\LaserSharpNT\Database\lsMessages.mdb.  If you have
> microsoft access, you can open a COPY of this file and look at the table
> called LaserSharpUsers and you can see all the user passwords in plain
> text.  Access level 0 is for system.  It is probably overly cautious to
> open a copy because the database will likely be opened as read-only; but
> why take a chance.
>
> Once you log in as an user with system access level, you should be able to
> change all the configuration parameters.  But be very careful, you can
> mess up things really bad.  I've learned that the hard way.  Hope this
> help.
>
> On Tue, 24 Mar 2009, Matiar Jafari wrote:
>
>> Dear all,
>>
>> So according to Chan the limit is hardcoded and cannot be changed....
>>
>> we have multiple accounts on the computer and im wondering if im not
>> able to change he memory option from 700 mb to the 1gb because its
>> hardcoded or because i need the "admin" login user for lasersharp
>> 2000....help? also when i change the directory for the temp files it
>> doesn't allow me to click ok thus it doesnt change where its
>> located....any help???
>>
>> On Tue, Mar 24, 2009 at 11:12 AM, Keith Morris <[hidden email]>
>> wrote:
>> > Hi Matiar,
>> > Chang and others have beaten me too it but anyway:
>> >
>> > I've had this error before on our Radiance 2000 and it was fixed pretty
>> > easily and permanently after a chat to the Bio-Rad engineer, but it was
>> > sorted out years ago so I am rather fuzzy on the reason for it and the
>> > cure.
>> > I'm sure though it is a memory error of some sort [I assume you are
>> > still
>> > running LaserSharp 2000 under NT - our error wasn't on the latest
>> > version 6
>> > but earlier, probably v4]. We did add more physical memory - to 768Mb or
>> > possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
>> > matched pairs], plus we played around with the hard drive space, system
>> > page
>> > file, LaserSharp settings logged in as system, and we put the crucial
>> > :\temp
>> > on our very much larger D:\ drive [set via Lasersharp system] as the C:\
>> > drive is always compromised to ~4Gb under NT, unless you increased it
>> > after
>> > SP6 is applied. We had two hard drives fitted, the first split into two
>> > partitions C:,E: and the much larger second one D: for user data and the
>> > Temp folder
>> >
>> > So I'm sure all the other postings are on the right track, it's system
>> > or
>> > hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what
>> > error
>> > message we got though other than the Z-stacks/time-lapses never finished
>> > and
>> > we lost everything]. I think we still had to split very long time-lapses
>> > with z stacks though into two runs [2Gb limit]. Also allocated more
>> > virtual
>> > memory [page file] from the hard drive to help out the physical RAM
>> > [forgotten how to do that under NT, but it's probably under 'system']. I
>> > think there used to be a 2Gb limit on collections though even with
>> > LaserSharp 6 [1Gb default] even with the tweeks.
>> >
>> > Zeiss [Microscience] should still support the Radiance 2000 [not the
>> > 1024
>> > though] so an email to their service support should clarify the
>> > solution. I
>> > don't know if upgrading to XP ever helped [you needed a new Firewire
>> > card
>> > and deep pockets for Bio-Rad to do this for you].
>> >
>> > Again look at the old postings below that seem very relevant.
>> >
>> > Keith
>> >
>> >
>> > ---------------------------------------------------------------------------
>> > Dr Keith J. Morris,
>> > Molecular Cytogenetics and Microscopy Core,
>> > Laboratory 00/069 and 00/070,
>> > The Wellcome Trust Centre for Human Genetics,
>> > Roosevelt Drive,
>> > Oxford  OX3 7BN,
>> > United Kingdom.
>> >
>> > Telephone:  +44 (0)1865 287568
>> > Email:  [hidden email]
>> > Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>> >
>> >
>> >
>> >
>> >
>> >
>> >
>> > -------------------------------------------------------------------
>> > Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
>> > From: W. Chan
>> > Reply-To: Confocal Microscopy List <[hidden email]>
>> > Date: Tue, 21 Mar 2006 16:59:39 -0800
>> > Content-Type: TEXT/PLAIN
>> >
>> > Search the CONFOCAL archive at
>> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>> >
>> > The Control Panel error message is quite misleading.  For v5.2, it
>> > states
>> > that "Collection Requires Too Much Memory" when the predicted dataset
>> > size
>> > exceeds the collection limit, but in fact it refers to disk space.
>> >
>> > I agree.  Even a 2 GB limit is really not much when you need
>> > multi-channels with a merge pane at 1024x1024 with multiple time points.
>> > May be a work around is not to have a merge pane which can be generated
>> > quite easily after you have the data.  A few of our users do not use
>> > the merge pane option in their methods.  They do not use LaserSharp to
>> > analyze their images so they can't read the merge pane .pmf file even if
>> > they want to.
>> >
>> > --
>> > Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>> > The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>> >
>> > On Sat, 18 Mar 2006, Michael Cammer wrote:
>> >
>> >> I'll check the upgrade software for writing to disk.  But with multiple
>> >> probe files our problem was the merge file.  For two probes, it needed
>> >> 150% more memory than the raw images themselves.  Even changing the
>> >> configuration didn't provide enough memory.
>> >>
>> >>
>> >>> Are you referring to RAM or harddisk space?  As far as I know,
>> > LaserSharps
>> >>> writes to temporary files rather than holding the experiment in RAM.
>> >>>  We
>> >>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with
>> >>> LaserSharp
>> >>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
>> >>> "System Configuration | Scan system" for the setting of the collection
>> >>> limit.  Previous versions may have a 1 GB limit.  Also check the
>> >>> location
>> >>> of the temporary files which needs to have plenty of space (as much as
>> > you
>> >>> want to acquire each time or more).  Good luck.
>> >>>
>> >>> --
>> >>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>> >>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>> >>>
>> >>> On Fri, 17 Mar 2006, Michael Cammer wrote:
>> >>>
>> >>>> We have the Radiance 2000.  This is a major limitation of the system.
>> >>>> We've had to compromise by restarting long time lapse experiments in
>> >>>> the
>> >>>> middle, by taking fewer Z planes per timepoint or longer intervals.
>> >>>> But when our BioRad confocals die, LaserSharp software should be
>> >>>> obsolete.
>> >>>> -Michael
>> >>>>
>> >>>> At 05:51 PM 03/14/06 -0800, you wrote:
>> >>>>>
>> >>>>> I have a question regarding a BioRad 1024 confocal system.  This
>> >>>>> system
>> >>>>> is
>> >>>>> working fine.  But one trouble is that people cannot run any
>> >>>>> scanning
>> >>>>> program (time lapse experiments) that contains more than 230 mb,
>> >>>>> although
>> >>>>> the system has plenty of free memory (we just added another 1gb to
>> >>>>> it).
>> >>>>>  I
>> >>>>> will appreciate if anyone can share with us if you had same
>> >>>>> experience
>> >>>>> and
>> >>>>> managed to resolve it.
>> >
>> > RECENT POSTINGS
>> > -----Original Message-----
>> > From: Confocal Microscopy List [mailto:[hidden email]]
>> > On
>> > Behalf Of Michael Bastiani
>> > Sent: 24 March 2009 13:57
>> > To: [hidden email]
>> > Subject: Re: LaserSharp 2000 Error
>> >
>> > Try increasing the memory allocated to image acquisition.  In Tools
>> > select System Config and set memory to size smaller than Computer
>> > RAM.  Also make sure your computer hard drive has plenty of space.
>> > You cannot collect images/stacks greater than the number set in System
>> > Config.
>> > good luck,
>> > michael
>> >
>> >
>> > On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
>> >
>> >> I've been getting this error when i shoot larger z stacks and when i
>> >> shoot longer time series
>> >>
>> >> Serious Error: NO file could be found matching the specification
>> >> 'C:\Temp\untitled\untitled1_raw*.pic'
>> >>
>> >> then i either click ok or close the dialog box and lasersharp 2000
>> >> closes/crashes
>> >>
>> >> but when i shoot smaller stacks this does NOT happen any help would be
>> >> greatly appreciated
>> >>
>> >> ive tried deleting the C:\Temp folder and this does nothing
>> >>
>> >> The reason i need this to finish is so that i can then export it to my
>> >> desired location on a different server.
>> >>
>> >> Thank You
>> >> ---
>> >> Matiar Jafari
>
> --
>
> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>
>
>
>



--
Matiar Jafari
Matiar Jafari Matiar Jafari
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

So i finally got LaserSharp 2000 to work with the help of a Zeiss
Representative and apparently theres a bug in v6 that does this two
work arounds one is to open another channel and turn off the laser the
other is to get the service pack.

Thanks for all the input!!!

On Fri, Mar 27, 2009 at 11:41 AM, Matiar Jafari <[hidden email]> wrote:

> update... my apologies i made a mistake im running windows 2000 with
> Lasersharp 2000 6.0 build 846 i just shot a stack that was only 19 um
> and it gave me the same error.
>
> I was told the work around was to
>
> "....the only solution is to use a method with 2 channels to collect
> data.  This will allow you to collect
> up to the limits set by LaserSharp ~2GB.  Once you have your data you
> can delete the empty/redundant channel of data."  Zeiss Support
>
> im shooting with a green method with only one channel any help in how
> i would do this?
>
> thanks in advance
>
>
> Thank You
>
> On Fri, Mar 27, 2009 at 2:42 AM, Keith Morris <[hidden email]> wrote:
>> More memory with NT/Lasersharp only helps if you are really down at the
>> 256Mb level, and mostly it gives stability/speed provided the modules are
>> matched [all ECC] because you probably still have that 1Gb to 2Gb upper
>> limit built into Lasersharp. Besides the hard drive page-file can [slowly]
>> provide more memory. Check your PC motherboard can cope with 2Gb physical
>> RAM [older systems have 1Gb and even 512Mb limits] – crucial.com often gives
>> this info. Don’t know if you have a MRC1024 or Radiance, but Zeiss
>> [Microscience] should be able to advise if your error messages are
>> consistent with Lasersharp v4/v5 operating normally or it’s an error [still
>> sound like its hitting the Lasersharp image capture memory limit of 1Gb or
>> 2Gb depending on version – unless of course you used to be able to capture
>> larger z-stacks]. Once you hit that Lasersharp limit, it’s either an upgrade
>> or you have to break the stack into a couple of separate ones. From memory
>> when an error message talks specifically about ‘not enough memory for
>> capture’ it frequently means hard drive space not system memory.
>>
>>
>>
>> Our Bio-Rad 1024 was no longer fully supported by Zeiss Micro-Science a few
>> years ago now and so we passed it on to those less fortunate and bought a
>> Zeiss 510 Meta. Zeiss did say they would continue to repair the MRC 1024
>> system while they could still get parts, and that they would continue to
>> provide technical support advice [which should be free via telephone/email].
>>
>>
>>
>> Many Bio-Rad staff have obviously left the company since UK Bio-Rad confocal
>> production was stopped by Zeiss, although they retained core personal for
>> product support of the newer Radiance 2000 confocals that also run
>> LaserSharp - some of whom will still remember the elderly 1024 [and perhaps
>> even COMOS, let alone LaserSharp]. Plus all the knowledgeable Bio-Rad
>> confocal reps generally live on working for other microscope companies.
>>
>>
>>
>> Keith
>>
>> ---------------------------------------------------------------------------
>> Dr Keith J. Morris,
>> Molecular Cytogenetics and Microscopy Core,
>> Laboratory 00/069 and 00/070,
>> The Wellcome Trust Centre for Human Genetics,
>> Roosevelt Drive,
>> Oxford  OX3 7BN,
>> United Kingdom.
>>
>> Telephone:  +44 (0)1865 287568
>> Email:  [hidden email]
>> Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>>
>> New error message after the first one I get. Failed to load PicMat with data
>> fileException in channels, stop capture
>>
>>
>>
>> I changed the ram capacity to 2gb and I tries changing the harddrive temp.
>>
>>
>>
>> I'm shooting then exporting but I can't export as it gives me the error
>> after I shoot anything larger than~80um
>>
>> Thank You
>>
>> ---
>>
>> Matiar Jafari
>>
>> On Mar 25, 2009, at 2:51 AM, Prof Hindsight <[hidden email]>
>> wrote:
>>
>> Dear Matiar Jafari,
>>
>>
>>
>> Have you actually contacted a Carl Zeiss service representative about this
>> problem?
>>
>>
>>
>> From my experience and that of others, Zeiss will assist with such
>> problems.  I also have it on good authority that although one can no longer
>> buy a service/maintenance contract for an MRC-1024, they will endeavour to
>> service the system on a billable basis for on-site work and, as Bio-Rad did
>> in the past, offer remote assistance at no cost.
>>
>> Whether you use Zeiss for support or not, knowing a bit more about what type
>> of confocal you actually have and its configuration would be helpful and
>> avoid misdiagnosis.
>>
>>
>>
>> Good luck,
>>
>>
>>
>> PH
>>
>> On Wed, Mar 25, 2009 at 1:24 AM, W. Chan <[hidden email]> wrote:
>>
>> Hi Matiar
>>
>> What I meant was e.g., in pre-5.2, you can only give max 1 GB to the
>> application.  It is not RAM related.  Since you did not mention what
>> version and OS you're running, I just assumed you're still using NT.
>> LaserSharp 6 and XP run really well together; but that's another story.  I
>> only have access to v6 now so the terms in the menu may be different than
>> earlier version.
>>
>> System access in lasersharp is completely independent of NT's (or whatever
>> window's) user privilege.  Usually, the software is set up with an user
>> called Administrator that have system access, other users for the software
>> may only have read & write access.  After you log into lasersharp, go to
>> Tools | User Setup, the second column will tell you what Access level you
>> have.  Whoever install the software should have the password to log in as
>> Administrator.
>>
>> There is a way to find that out.  Everything for this software are stored
>> in a database e.g., c:\LaserSharpNT\Database\lsMessages.mdb.  If you have
>> microsoft access, you can open a COPY of this file and look at the table
>> called LaserSharpUsers and you can see all the user passwords in plain
>> text.  Access level 0 is for system.  It is probably overly cautious to
>> open a copy because the database will likely be opened as read-only; but
>> why take a chance.
>>
>> Once you log in as an user with system access level, you should be able to
>> change all the configuration parameters.  But be very careful, you can
>> mess up things really bad.  I've learned that the hard way.  Hope this
>> help.
>>
>> On Tue, 24 Mar 2009, Matiar Jafari wrote:
>>
>>> Dear all,
>>>
>>> So according to Chan the limit is hardcoded and cannot be changed....
>>>
>>> we have multiple accounts on the computer and im wondering if im not
>>> able to change he memory option from 700 mb to the 1gb because its
>>> hardcoded or because i need the "admin" login user for lasersharp
>>> 2000....help? also when i change the directory for the temp files it
>>> doesn't allow me to click ok thus it doesnt change where its
>>> located....any help???
>>>
>>> On Tue, Mar 24, 2009 at 11:12 AM, Keith Morris <[hidden email]>
>>> wrote:
>>> > Hi Matiar,
>>> > Chang and others have beaten me too it but anyway:
>>> >
>>> > I've had this error before on our Radiance 2000 and it was fixed pretty
>>> > easily and permanently after a chat to the Bio-Rad engineer, but it was
>>> > sorted out years ago so I am rather fuzzy on the reason for it and the
>>> > cure.
>>> > I'm sure though it is a memory error of some sort [I assume you are
>>> > still
>>> > running LaserSharp 2000 under NT - our error wasn't on the latest
>>> > version 6
>>> > but earlier, probably v4]. We did add more physical memory - to 768Mb or
>>> > possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
>>> > matched pairs], plus we played around with the hard drive space, system
>>> > page
>>> > file, LaserSharp settings logged in as system, and we put the crucial
>>> > :\temp
>>> > on our very much larger D:\ drive [set via Lasersharp system] as the C:\
>>> > drive is always compromised to ~4Gb under NT, unless you increased it
>>> > after
>>> > SP6 is applied. We had two hard drives fitted, the first split into two
>>> > partitions C:,E: and the much larger second one D: for user data and the
>>> > Temp folder
>>> >
>>> > So I'm sure all the other postings are on the right track, it's system
>>> > or
>>> > hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what
>>> > error
>>> > message we got though other than the Z-stacks/time-lapses never finished
>>> > and
>>> > we lost everything]. I think we still had to split very long time-lapses
>>> > with z stacks though into two runs [2Gb limit]. Also allocated more
>>> > virtual
>>> > memory [page file] from the hard drive to help out the physical RAM
>>> > [forgotten how to do that under NT, but it's probably under 'system']. I
>>> > think there used to be a 2Gb limit on collections though even with
>>> > LaserSharp 6 [1Gb default] even with the tweeks.
>>> >
>>> > Zeiss [Microscience] should still support the Radiance 2000 [not the
>>> > 1024
>>> > though] so an email to their service support should clarify the
>>> > solution. I
>>> > don't know if upgrading to XP ever helped [you needed a new Firewire
>>> > card
>>> > and deep pockets for Bio-Rad to do this for you].
>>> >
>>> > Again look at the old postings below that seem very relevant.
>>> >
>>> > Keith
>>> >
>>> >
>>> > ---------------------------------------------------------------------------
>>> > Dr Keith J. Morris,
>>> > Molecular Cytogenetics and Microscopy Core,
>>> > Laboratory 00/069 and 00/070,
>>> > The Wellcome Trust Centre for Human Genetics,
>>> > Roosevelt Drive,
>>> > Oxford  OX3 7BN,
>>> > United Kingdom.
>>> >
>>> > Telephone:  +44 (0)1865 287568
>>> > Email:  [hidden email]
>>> > Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>>> >
>>> >
>>> >
>>> >
>>> >
>>> >
>>> >
>>> > -------------------------------------------------------------------
>>> > Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
>>> > From: W. Chan
>>> > Reply-To: Confocal Microscopy List <[hidden email]>
>>> > Date: Tue, 21 Mar 2006 16:59:39 -0800
>>> > Content-Type: TEXT/PLAIN
>>> >
>>> > Search the CONFOCAL archive at
>>> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>> >
>>> > The Control Panel error message is quite misleading.  For v5.2, it
>>> > states
>>> > that "Collection Requires Too Much Memory" when the predicted dataset
>>> > size
>>> > exceeds the collection limit, but in fact it refers to disk space.
>>> >
>>> > I agree.  Even a 2 GB limit is really not much when you need
>>> > multi-channels with a merge pane at 1024x1024 with multiple time points.
>>> > May be a work around is not to have a merge pane which can be generated
>>> > quite easily after you have the data.  A few of our users do not use
>>> > the merge pane option in their methods.  They do not use LaserSharp to
>>> > analyze their images so they can't read the merge pane .pmf file even if
>>> > they want to.
>>> >
>>> > --
>>> > Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>>> > The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>>> >
>>> > On Sat, 18 Mar 2006, Michael Cammer wrote:
>>> >
>>> >> I'll check the upgrade software for writing to disk.  But with multiple
>>> >> probe files our problem was the merge file.  For two probes, it needed
>>> >> 150% more memory than the raw images themselves.  Even changing the
>>> >> configuration didn't provide enough memory.
>>> >>
>>> >>
>>> >>> Are you referring to RAM or harddisk space?  As far as I know,
>>> > LaserSharps
>>> >>> writes to temporary files rather than holding the experiment in RAM.
>>> >>>  We
>>> >>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with
>>> >>> LaserSharp
>>> >>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
>>> >>> "System Configuration | Scan system" for the setting of the collection
>>> >>> limit.  Previous versions may have a 1 GB limit.  Also check the
>>> >>> location
>>> >>> of the temporary files which needs to have plenty of space (as much as
>>> > you
>>> >>> want to acquire each time or more).  Good luck.
>>> >>>
>>> >>> --
>>> >>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>>> >>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>>> >>>
>>> >>> On Fri, 17 Mar 2006, Michael Cammer wrote:
>>> >>>
>>> >>>> We have the Radiance 2000.  This is a major limitation of the system.
>>> >>>> We've had to compromise by restarting long time lapse experiments in
>>> >>>> the
>>> >>>> middle, by taking fewer Z planes per timepoint or longer intervals.
>>> >>>> But when our BioRad confocals die, LaserSharp software should be
>>> >>>> obsolete.
>>> >>>> -Michael
>>> >>>>
>>> >>>> At 05:51 PM 03/14/06 -0800, you wrote:
>>> >>>>>
>>> >>>>> I have a question regarding a BioRad 1024 confocal system.  This
>>> >>>>> system
>>> >>>>> is
>>> >>>>> working fine.  But one trouble is that people cannot run any
>>> >>>>> scanning
>>> >>>>> program (time lapse experiments) that contains more than 230 mb,
>>> >>>>> although
>>> >>>>> the system has plenty of free memory (we just added another 1gb to
>>> >>>>> it).
>>> >>>>>  I
>>> >>>>> will appreciate if anyone can share with us if you had same
>>> >>>>> experience
>>> >>>>> and
>>> >>>>> managed to resolve it.
>>> >
>>> > RECENT POSTINGS
>>> > -----Original Message-----
>>> > From: Confocal Microscopy List [mailto:[hidden email]]
>>> > On
>>> > Behalf Of Michael Bastiani
>>> > Sent: 24 March 2009 13:57
>>> > To: [hidden email]
>>> > Subject: Re: LaserSharp 2000 Error
>>> >
>>> > Try increasing the memory allocated to image acquisition.  In Tools
>>> > select System Config and set memory to size smaller than Computer
>>> > RAM.  Also make sure your computer hard drive has plenty of space.
>>> > You cannot collect images/stacks greater than the number set in System
>>> > Config.
>>> > good luck,
>>> > michael
>>> >
>>> >
>>> > On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
>>> >
>>> >> I've been getting this error when i shoot larger z stacks and when i
>>> >> shoot longer time series
>>> >>
>>> >> Serious Error: NO file could be found matching the specification
>>> >> 'C:\Temp\untitled\untitled1_raw*.pic'
>>> >>
>>> >> then i either click ok or close the dialog box and lasersharp 2000
>>> >> closes/crashes
>>> >>
>>> >> but when i shoot smaller stacks this does NOT happen any help would be
>>> >> greatly appreciated
>>> >>
>>> >> ive tried deleting the C:\Temp folder and this does nothing
>>> >>
>>> >> The reason i need this to finish is so that i can then export it to my
>>> >> desired location on a different server.
>>> >>
>>> >> Thank You
>>> >> ---
>>> >> Matiar Jafari
>>
>> --
>>
>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>>
>>
>>
>>
>
>
>
> --
> Matiar Jafari
>



--
Matiar Jafari
Keith Morris Keith Morris
Reply | Threaded
Open this post in threaded view
|

Re: LaserSharp 2000 Error

That was a problem with Bio-Rad's software, bugs. Every Lasersharp version
seemed to start off with one or two major ones - the British software
writers tended to get the new version out ASAP and then issue patches ASAP
[which naturally you need to apply]. As v6 is the last and best Lasersharp,
I expect Zeiss with give you the most up to date version for free.

The methodical Germans [e.g. Zeiss] tend to bug the test the software for
months or years before releasing it [so neat new features took a longer time
to appear]. Given the small user base of the LSM software they all do pretty
well though [even Adobe software is pretty bug ridden on first release].

Keith  

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford  OX3 7BN,
United Kingdom.

Telephone:  +44 (0)1865 287568
Email:  [hidden email]
Web-pages: http://www.well.ox.ac.uk/cytogenetics/
 

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Matiar Jafari
Sent: 28 March 2009 00:44
To: [hidden email]
Subject: Re: LaserSharp 2000 Error

So i finally got LaserSharp 2000 to work with the help of a Zeiss
Representative and apparently theres a bug in v6 that does this two
work arounds one is to open another channel and turn off the laser the
other is to get the service pack.

Thanks for all the input!!!

On Fri, Mar 27, 2009 at 11:41 AM, Matiar Jafari <[hidden email]>
wrote:

> update... my apologies i made a mistake im running windows 2000 with
> Lasersharp 2000 6.0 build 846 i just shot a stack that was only 19 um
> and it gave me the same error.
>
> I was told the work around was to
>
> "....the only solution is to use a method with 2 channels to collect
> data.  This will allow you to collect
> up to the limits set by LaserSharp ~2GB.  Once you have your data you
> can delete the empty/redundant channel of data."  Zeiss Support
>
> im shooting with a green method with only one channel any help in how
> i would do this?
>
> thanks in advance
>
>
> Thank You
>
> On Fri, Mar 27, 2009 at 2:42 AM, Keith Morris <[hidden email]>
wrote:
>> More memory with NT/Lasersharp only helps if you are really down at the
>> 256Mb level, and mostly it gives stability/speed provided the modules are
>> matched [all ECC] because you probably still have that 1Gb to 2Gb upper
>> limit built into Lasersharp. Besides the hard drive page-file can
[slowly]
>> provide more memory. Check your PC motherboard can cope with 2Gb physical
>> RAM [older systems have 1Gb and even 512Mb limits] – crucial.com often
gives
>> this info. Don’t know if you have a MRC1024 or Radiance, but Zeiss
>> [Microscience] should be able to advise if your error messages are
>> consistent with Lasersharp v4/v5 operating normally or it’s an error
[still
>> sound like its hitting the Lasersharp image capture memory limit of 1Gb
or
>> 2Gb depending on version – unless of course you used to be able to
capture
>> larger z-stacks]. Once you hit that Lasersharp limit, it’s either an
upgrade
>> or you have to break the stack into a couple of separate ones. From
memory
>> when an error message talks specifically about ‘not enough memory for
>> capture’ it frequently means hard drive space not system memory.
>>
>>
>>
>> Our Bio-Rad 1024 was no longer fully supported by Zeiss Micro-Science a
few
>> years ago now and so we passed it on to those less fortunate and bought a
>> Zeiss 510 Meta. Zeiss did say they would continue to repair the MRC 1024
>> system while they could still get parts, and that they would continue to
>> provide technical support advice [which should be free via
telephone/email].
>>
>>
>>
>> Many Bio-Rad staff have obviously left the company since UK Bio-Rad
confocal
>> production was stopped by Zeiss, although they retained core personal for
>> product support of the newer Radiance 2000 confocals that also run
>> LaserSharp - some of whom will still remember the elderly 1024 [and
perhaps
>> even COMOS, let alone LaserSharp]. Plus all the knowledgeable Bio-Rad
>> confocal reps generally live on working for other microscope companies.
>>
>>
>>
>> Keith
>>
>>
---------------------------------------------------------------------------

>> Dr Keith J. Morris,
>> Molecular Cytogenetics and Microscopy Core,
>> Laboratory 00/069 and 00/070,
>> The Wellcome Trust Centre for Human Genetics,
>> Roosevelt Drive,
>> Oxford  OX3 7BN,
>> United Kingdom.
>>
>> Telephone:  +44 (0)1865 287568
>> Email:  [hidden email]
>> Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>>
>> New error message after the first one I get. Failed to load PicMat with
data
>> fileException in channels, stop capture
>>
>>
>>
>> I changed the ram capacity to 2gb and I tries changing the harddrive
temp.

>>
>>
>>
>> I'm shooting then exporting but I can't export as it gives me the error
>> after I shoot anything larger than~80um
>>
>> Thank You
>>
>> ---
>>
>> Matiar Jafari
>>
>> On Mar 25, 2009, at 2:51 AM, Prof Hindsight
<[hidden email]>
>> wrote:
>>
>> Dear Matiar Jafari,
>>
>>
>>
>> Have you actually contacted a Carl Zeiss service representative about
this
>> problem?
>>
>>
>>
>> From my experience and that of others, Zeiss will assist with such
>> problems.  I also have it on good authority that although one can no
longer
>> buy a service/maintenance contract for an MRC-1024, they will endeavour
to
>> service the system on a billable basis for on-site work and, as Bio-Rad
did
>> in the past, offer remote assistance at no cost.
>>
>> Whether you use Zeiss for support or not, knowing a bit more about what
type

>> of confocal you actually have and its configuration would be helpful and
>> avoid misdiagnosis.
>>
>>
>>
>> Good luck,
>>
>>
>>
>> PH
>>
>> On Wed, Mar 25, 2009 at 1:24 AM, W. Chan <[hidden email]> wrote:
>>
>> Hi Matiar
>>
>> What I meant was e.g., in pre-5.2, you can only give max 1 GB to the
>> application.  It is not RAM related.  Since you did not mention what
>> version and OS you're running, I just assumed you're still using NT.
>> LaserSharp 6 and XP run really well together; but that's another story.
 I
>> only have access to v6 now so the terms in the menu may be different than
>> earlier version.
>>
>> System access in lasersharp is completely independent of NT's (or
whatever
>> window's) user privilege.  Usually, the software is set up with an user
>> called Administrator that have system access, other users for the
software

>> may only have read & write access.  After you log into lasersharp, go to
>> Tools | User Setup, the second column will tell you what Access level you
>> have.  Whoever install the software should have the password to log in as
>> Administrator.
>>
>> There is a way to find that out.  Everything for this software are stored
>> in a database e.g., c:\LaserSharpNT\Database\lsMessages.mdb.  If you have
>> microsoft access, you can open a COPY of this file and look at the table
>> called LaserSharpUsers and you can see all the user passwords in plain
>> text.  Access level 0 is for system.  It is probably overly cautious to
>> open a copy because the database will likely be opened as read-only; but
>> why take a chance.
>>
>> Once you log in as an user with system access level, you should be able
to

>> change all the configuration parameters.  But be very careful, you can
>> mess up things really bad.  I've learned that the hard way.  Hope this
>> help.
>>
>> On Tue, 24 Mar 2009, Matiar Jafari wrote:
>>
>>> Dear all,
>>>
>>> So according to Chan the limit is hardcoded and cannot be changed....
>>>
>>> we have multiple accounts on the computer and im wondering if im not
>>> able to change he memory option from 700 mb to the 1gb because its
>>> hardcoded or because i need the "admin" login user for lasersharp
>>> 2000....help? also when i change the directory for the temp files it
>>> doesn't allow me to click ok thus it doesnt change where its
>>> located....any help???
>>>
>>> On Tue, Mar 24, 2009 at 11:12 AM, Keith Morris <[hidden email]>
>>> wrote:
>>> > Hi Matiar,
>>> > Chang and others have beaten me too it but anyway:
>>> >
>>> > I've had this error before on our Radiance 2000 and it was fixed
pretty
>>> > easily and permanently after a chat to the Bio-Rad engineer, but it
was
>>> > sorted out years ago so I am rather fuzzy on the reason for it and the
>>> > cure.
>>> > I'm sure though it is a memory error of some sort [I assume you are
>>> > still
>>> > running LaserSharp 2000 under NT - our error wasn't on the latest
>>> > version 6
>>> > but earlier, probably v4]. We did add more physical memory - to 768Mb
or
>>> > possibly 1Gb [try eBay for cheap s/h ECC memory, and check if you need
>>> > matched pairs], plus we played around with the hard drive space,
system
>>> > page
>>> > file, LaserSharp settings logged in as system, and we put the crucial
>>> > :\temp
>>> > on our very much larger D:\ drive [set via Lasersharp system] as the
C:\
>>> > drive is always compromised to ~4Gb under NT, unless you increased it
>>> > after
>>> > SP6 is applied. We had two hard drives fitted, the first split into
two
>>> > partitions C:,E: and the much larger second one D: for user data and
the
>>> > Temp folder
>>> >
>>> > So I'm sure all the other postings are on the right track, it's system
>>> > or
>>> > hard drive memory and the 1Gb/2Gb limit - see Pangs post [no idea what
>>> > error
>>> > message we got though other than the Z-stacks/time-lapses never
finished
>>> > and
>>> > we lost everything]. I think we still had to split very long
time-lapses
>>> > with z stacks though into two runs [2Gb limit]. Also allocated more
>>> > virtual
>>> > memory [page file] from the hard drive to help out the physical RAM
>>> > [forgotten how to do that under NT, but it's probably under 'system'].
I

>>> > think there used to be a 2Gb limit on collections though even with
>>> > LaserSharp 6 [1Gb default] even with the tweeks.
>>> >
>>> > Zeiss [Microscience] should still support the Radiance 2000 [not the
>>> > 1024
>>> > though] so an email to their service support should clarify the
>>> > solution. I
>>> > don't know if upgrading to XP ever helped [you needed a new Firewire
>>> > card
>>> > and deep pockets for Bio-Rad to do this for you].
>>> >
>>> > Again look at the old postings below that seem very relevant.
>>> >
>>> > Keith
>>> >
>>> >
>>> >
---------------------------------------------------------------------------

>>> > Dr Keith J. Morris,
>>> > Molecular Cytogenetics and Microscopy Core,
>>> > Laboratory 00/069 and 00/070,
>>> > The Wellcome Trust Centre for Human Genetics,
>>> > Roosevelt Drive,
>>> > Oxford  OX3 7BN,
>>> > United Kingdom.
>>> >
>>> > Telephone:  +44 (0)1865 287568
>>> > Email:  [hidden email]
>>> > Web-pages: http://www.well.ox.ac.uk/cytogenetics/
>>> >
>>> >
>>> >
>>> >
>>> >
>>> >
>>> >
>>> > -------------------------------------------------------------------
>>> > Subject: Re: Radiance 2000 (was Re: [CONFOCAL] Bio-Rad 1024)
>>> > From: W. Chan
>>> > Reply-To: Confocal Microscopy List <[hidden email]>
>>> > Date: Tue, 21 Mar 2006 16:59:39 -0800
>>> > Content-Type: TEXT/PLAIN
>>> >
>>> > Search the CONFOCAL archive at
>>> > http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>> >
>>> > The Control Panel error message is quite misleading.  For v5.2, it
>>> > states
>>> > that "Collection Requires Too Much Memory" when the predicted dataset
>>> > size
>>> > exceeds the collection limit, but in fact it refers to disk space.
>>> >
>>> > I agree.  Even a 2 GB limit is really not much when you need
>>> > multi-channels with a merge pane at 1024x1024 with multiple time
points.
>>> > May be a work around is not to have a merge pane which can be
generated
>>> > quite easily after you have the data.  A few of our users do not use
>>> > the merge pane option in their methods.  They do not use LaserSharp to
>>> > analyze their images so they can't read the merge pane .pmf file even
if
>>> > they want to.
>>> >
>>> > --
>>> > Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>>> > The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>>> >
>>> > On Sat, 18 Mar 2006, Michael Cammer wrote:
>>> >
>>> >> I'll check the upgrade software for writing to disk.  But with
multiple
>>> >> probe files our problem was the merge file.  For two probes, it
needed

>>> >> 150% more memory than the raw images themselves.  Even changing the
>>> >> configuration didn't provide enough memory.
>>> >>
>>> >>
>>> >>> Are you referring to RAM or harddisk space?  As far as I know,
>>> > LaserSharps
>>> >>> writes to temporary files rather than holding the experiment in RAM.
>>> >>>  We
>>> >>> have a Radiance 2000 on NT (1 GB RAM, 1.5 GB page file) with
>>> >>> LaserSharp
>>> >>> 5.2 build 824.  The collection limit is 2 GB.  You may want to check
>>> >>> "System Configuration | Scan system" for the setting of the
collection
>>> >>> limit.  Previous versions may have a 1 GB limit.  Also check the
>>> >>> location
>>> >>> of the temporary files which needs to have plenty of space (as much
as
>>> > you
>>> >>> want to acquire each time or more).  Good luck.
>>> >>>
>>> >>> --
>>> >>> Pang (Wai Pang Chan, [hidden email], PAB A087,
206-685-1519)
>>> >>> The Biology Imaging Facility
(http://staff.washington.edu/wpchan/if/)
>>> >>>
>>> >>> On Fri, 17 Mar 2006, Michael Cammer wrote:
>>> >>>
>>> >>>> We have the Radiance 2000.  This is a major limitation of the
system.
>>> >>>> We've had to compromise by restarting long time lapse experiments
in

>>> >>>> the
>>> >>>> middle, by taking fewer Z planes per timepoint or longer intervals.
>>> >>>> But when our BioRad confocals die, LaserSharp software should be
>>> >>>> obsolete.
>>> >>>> -Michael
>>> >>>>
>>> >>>> At 05:51 PM 03/14/06 -0800, you wrote:
>>> >>>>>
>>> >>>>> I have a question regarding a BioRad 1024 confocal system.  This
>>> >>>>> system
>>> >>>>> is
>>> >>>>> working fine.  But one trouble is that people cannot run any
>>> >>>>> scanning
>>> >>>>> program (time lapse experiments) that contains more than 230 mb,
>>> >>>>> although
>>> >>>>> the system has plenty of free memory (we just added another 1gb to
>>> >>>>> it).
>>> >>>>>  I
>>> >>>>> will appreciate if anyone can share with us if you had same
>>> >>>>> experience
>>> >>>>> and
>>> >>>>> managed to resolve it.
>>> >
>>> > RECENT POSTINGS
>>> > -----Original Message-----
>>> > From: Confocal Microscopy List
[mailto:[hidden email]]

>>> > On
>>> > Behalf Of Michael Bastiani
>>> > Sent: 24 March 2009 13:57
>>> > To: [hidden email]
>>> > Subject: Re: LaserSharp 2000 Error
>>> >
>>> > Try increasing the memory allocated to image acquisition.  In Tools
>>> > select System Config and set memory to size smaller than Computer
>>> > RAM.  Also make sure your computer hard drive has plenty of space.
>>> > You cannot collect images/stacks greater than the number set in System
>>> > Config.
>>> > good luck,
>>> > michael
>>> >
>>> >
>>> > On Mar 24, 2009, at 2:57 AM, Matiar Jafari wrote:
>>> >
>>> >> I've been getting this error when i shoot larger z stacks and when i
>>> >> shoot longer time series
>>> >>
>>> >> Serious Error: NO file could be found matching the specification
>>> >> 'C:\Temp\untitled\untitled1_raw*.pic'
>>> >>
>>> >> then i either click ok or close the dialog box and lasersharp 2000
>>> >> closes/crashes
>>> >>
>>> >> but when i shoot smaller stacks this does NOT happen any help would
be
>>> >> greatly appreciated
>>> >>
>>> >> ive tried deleting the C:\Temp folder and this does nothing
>>> >>
>>> >> The reason i need this to finish is so that i can then export it to
my

>>> >> desired location on a different server.
>>> >>
>>> >> Thank You
>>> >> ---
>>> >> Matiar Jafari
>>
>> --
>>
>> Pang (Wai Pang Chan, [hidden email], PAB A087, 206-685-1519)
>> The Biology Imaging Facility (http://staff.washington.edu/wpchan/if/)
>>
>>
>>
>>
>
>
>
> --
> Matiar Jafari
>



--
Matiar Jafari