Stability issues with LAS AF 1.8.0

[Nowell, Cameron]

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

Howdy List,
                 We have a leica SP5 system with the leica super Z peizo stage that we recently had upgraded to version 1.8.0 of LAS AF. After the upgrade we ran a test expeirment over night (multiple stage positions and different z stacks) and it ran perfectly. Since then it has crashed about 15-20 times in the sapce of 3 days. We had a new super z stage installed at the same time as the upgrade (to correct some drift issues we were having). But i do not think the crashing is due to the super z stage because the software will crash when you use the zoom functions as well or use wide focus instead of galvo.

Is anyone out there running version 1.8.0 of LAS AF and if so have you had similar issues. We have now downgraded the software back to 1.7.0 and it seems to be running smoothly again. I just need to know if this is a global problem with ver 1.8.0 or it is something specific to our instrument. The leica technitian took 5 goes to get version 1.8 installed as it kept failing at different parts of the firm ware upgrade process. Similarly when taking it back to ver 1.7 i ahd the same problems.

On another note does anyone out there with a super z stage have it vibrate on them? Both stages we have had vibrate and emit a high pitched wine. If you poke it it will stop and everthing is fine. If it is vibrating when you are trying to capture an image then you don't get a good picture.

Thanks

Cam

Cameron Nowell B. Sc (Hons)
Microscopy Imaging and Research Core Facility
Peter MacCallum Cancer Centre
7 St Andrews Place
East Melbourne, Victoria 3002
AUSTRALIA
Phone: [phone]
Mobile: [phone]
Fax: [phone]

This email (including any attachments) may contain
confidential and/or legally privileged information and is
intended only to be read or used by the addressee. If you
are not the intended addressee, any use, distribution,
disclosure or copying of this email is strictly
prohibited.
Confidentiality and legal privilege attached to this email
(including any attachments) are not waived or lost by
reason of its mistaken delivery to you.
If you have received this email in error, please delete it
and notify us immediately by telephone or email. Peter
MacCallum Cancer Centre provides no guarantee that this
transmission is free of virus or that it has not been
intercepted or altered and will not be liable for any delay
in its receipt.

[Dmitry Sokolov-2]

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

May I ask you for your suggestion on proper procedure of mercury arc bulb alignment of fiber-coupled 106 z lamp housing. I was recommended to align it by maximum of intensity of light passing through the objective of (Leica DM6000B) microscope . The intensity and uniformity is controlled looking on a sheet of paper placed under turret with objective removed.

The corresponding "Leica DM6000B DM6000M Operating manual" has a warning of lamp explosion by overheating if "The bright tips of the arcs, the focal sports..." projected onto each other (p.55) or contact elements of lamp. The alignment procedure is based on viewing the arcs through a special window of the microscope. In my case I see no images of arcs but a number of rings originated probably from the fiber ends.

Should I better remove the fiber coupler and align the lamp looking on it's image on the wall?

I would highly appreciate it if you could share your experience.

Cheers,
Dmitry

[Michael Weber-4]

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

Dear Cam,

we are running v1.8 since a couple of weeks and I have to say that this
is the first one which hasn't crashed so far. We had more problems with
v1.6 and v1.7. So it might be an electronics problem on your side. I
also know people still having the same crashing issues with every version.

We had the issue of vibrating stage once during the one year we have the
system, similar to your description. The maximum allowed load for this
stage is quite low (100g?), so I have the suspicion that this vibration
issue could be due to overloading. We also have an older SP2 with Galvo
Stage and this is out of calibration from time to time, probably also
because of overload - this one is an upright stand, so as soon as you
hit the sample with the objective it's for sure more than 100g...

cheers,
Michael


Nowell, Cameron wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Howdy List,
>                  We have a leica SP5 system with the leica super Z peizo
> stage that we recently had upgraded to version 1.8.0 of LAS AF. After
> the upgrade we ran a test expeirment over night (multiple stage
> positions and different z stacks) and it ran perfectly. Since then it
> has crashed about 15-20 times in the sapce of 3 days. We had a new super
> z stage installed at the same time as the upgrade (to correct some drift
> issues we were having). But i do not think the crashing is due to the
> super z stage because the software will crash when you use the zoom
> functions as well or use wide focus instead of galvo.
>
> Is anyone out there running version 1.8.0 of LAS AF and if so have you
> had similar issues. We have now downgraded the software back to 1.7.0
> and it seems to be running smoothly again. I just need to know if this
> is a global problem with ver 1.8.0 or it is something specific to our
> instrument. The leica technitian took 5 goes to get version 1.8
> installed as it kept failing at different parts of the firm ware upgrade
> process. Similarly when taking it back to ver 1.7 i ahd the same problems.
>
> On another note does anyone out there with a super z stage have it
> vibrate on them? Both stages we have had vibrate and emit a high pitched
> wine. If you poke it it will stop and everthing is fine. If it is
> vibrating when you are trying to capture an image then you don't get a
> good picture.
>
>
> Thanks
>
>
> Cam
>
>
> *Cameron Nowell* B. Sc (Hons)
> Microscopy Imaging and Research Core Facility
> Peter MacCallum Cancer Centre
> 7 St Andrews Place
> East Melbourne, Victoria 3002
> AUSTRALIA
> *Phone:* [phone]
> *Mobile:* [phone]
> *Fax:* [phone]
>
> This email (including any attachments) may contain
> confidential and/or legally privileged information and is
> intended only to be read or used by the addressee. If you
> are not the intended addressee, any use, distribution,
> disclosure or copying of this email is strictly
> prohibited.
> Confidentiality and legal privilege attached to this email
> (including any attachments) are not waived or lost by
> reason of its mistaken delivery to you.
> If you have received this email in error, please delete it
> and notify us immediately by telephone or email. Peter
> MacCallum Cancer Centre provides no guarantee that this
> transmission is free of virus or that it has not been
> intercepted or altered and will not be liable for any delay
> in its receipt.

[Michael Weber-4]

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

Dear Dmitry,

you won't be able to see the lamp filaments after passing through the
fiber. So yes, if you want to align the lamp in a proper way, you have
to remove the fiber from the lamphouse and align it at the wall (or an
another stand).

I have to say that I so far did it also based on highest throughput, but
I know about the security issues and will (hopefully) do it better soon.
The fiber is anyway killing a lot of light, so on a longterm view it
might be better to go for other solutions, i.e. LED sources or stronger
Metal Halide lamps with better fibers.

cheers,
Michael


Dmitry Sokolov wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear All,
>
> May I ask you for your suggestion on proper procedure of mercury arc
> bulb alignment of fiber-coupled 106 z lamp housing. I was recommended to
> align it by maximum of intensity of light passing through the objective
> of (Leica DM6000B) microscope . The intensity and uniformity is
> controlled looking on a sheet of paper placed under turret with
> objective removed.
>
> The corresponding "Leica DM6000B DM6000M Operating manual" has a warning
> of lamp explosion by overheating if "The bright tips of the arcs, the
> focal sports..." projected onto each other (p.55) or contact elements of
> lamp. The alignment procedure is based on viewing the arcs through a
> special window of the microscope. In my case I see no images of arcs but
> a number of rings originated probably from the fiber ends.
>
> Should I better remove the fiber coupler and align the lamp looking on
> it's image on the wall?
>
> I would highly appreciate it if you could share your experience.
>
> Cheers,
> Dmitry
>
> ___________________________________________
> Dr. Dmitry Sokolov,
> Manager of Confocal Microscopy Unit
> Institute of Molecular Biosciences
> Riddet Road, Massey University
> Private Bag 11-222, Palmerston North
> Tel: [phone] ext. 5549
> Email: [hidden email] <mailto:[hidden email]>
> Booking: http://confocal-manawatu.pbwiki.com/Booking

[Andrew Resnick]

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

Here's what I do, not sure if it's "factory approved":

First, align the bulb and reflected image of the bulb.  Do this by removing the fiber coupler and projecting the light on a far wall.  The two images of the bulb (direct and reflected) should both be in focus and side-by-side.  Do not overlap the bulb images!

Next, add the fiber coupler, and adjust it (the fiber mount) until the maximum amount of light exits the fiber.

Finally, do a final adjustment by attaching the fiber coupler to the microscope and one of the following:

1) focus onto a fluorescent slide (Chroma makes a nice set) and adjust the fiber/microscope coupling to get a uniform field of illumination
2) Use the special window you mention below.  A smoked glass side-looking mirror is in place in the fluorescent turret to reflect the illumination to the side port.
3) Project the illumination normally, but without an objective.  Adjust to get a uniform illumination pattern.

I prefer to use #1, as it's more sensible.
Hope this helps...

Andy

At 08:04 PM 11/13/2007, you wrote:

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

May I ask you for your suggestion on proper procedure of mercury arc bulb alignment of fiber-coupled 106 z lamp housing. I was recommended to align it by maximum of intensity of light passing through the objective of (Leica DM6000B) microscope . The intensity and uniformity is controlled looking on a sheet of paper placed under turret with objective removed.

The corresponding "Leica DM6000B DM6000M Operating manual" has a warning of lamp explosion by overheating if "The bright tips of the arcs, the focal sports..." projected onto each other (p.55) or contact elements of lamp. The alignment procedure is based on viewing the arcs through a special window of the microscope. In my case I see no images of arcs but a number of rings originated probably from the fiber ends.

Should I better remove the fiber coupler and align the lamp looking on it's image on the wall?

I would highly appreciate it if you could share your experience.

Cheers,
Dmitry

___________________________________________
Dr. Dmitry Sokolov,
Manager of Confocal Microscopy Unit
Institute of Molecular Biosciences
Riddet Road, Massey University
Private Bag 11-222, Palmerston North
Tel: [phone] ext. 5549
Email: [hidden email]
Booking: http://confocal-manawatu.pbwiki.com/Booking

Andrew Resnick, Ph. D.
Instructor
Department of Physiology and Biophysics
Case Western Reserve University
216-368-6899 (V)
216-368-4223 (F)

[Jean-Christophe Olaya]

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

Dear Cam,

Actually we have been having the unstability problem you decribe for  
two months.
To tell the complete story : just after we moved our two SP5 from one  
room to another, both of them started (on the same day) to crash every  
10-20 minute during scans.
We were running under the version 1.7.0 which was told not to be very  
stable, so we upgraded to version 1.8.0. Under this version the system  
was crashing, let's say, every hour. So we decided to downgrade to  
version 1.6.3 with which the system used to be much more stable.
Actually, the crash rate of our system is still something like one  
crash per hour, and we are still fighting to find the source of the  
problem, because it seems that it is not (only) a software version  
problem (otherwise it would not crash anymore with version 1.6.3, as  
it used to be).

All I can tell you is to check the following problem-causes we have  
been going through :

-disconnect your computer from any network and avoid having other  
software running paralelly to the LAS AF software : we have been told  
that antivirus updates during a scan may cause system to crash

-check if your system is on a stabilized power supply ; the scanhead  
may get crazy in case of fluctuations

-control the hardware tree of your system (or just send an error log  
to Leica service, they will control it) : in our case, the system was  
initialising looking for some non-descan system that we did not have

All these details may improve the stability of your system.
But we still did not manage to solve the problem completely from our  
side, so I would actually also be very interested in knowing if  
someone else is experiencing this kind of problem (and a solution  
would be very welcome...)

JC



"Nowell, Cameron" <[hidden email]> a écrit :

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Howdy List,
>                  We have a leica SP5 system with the leica super Z peizo
> stage that we recently had upgraded to version 1.8.0 of LAS AF. After
> the upgrade we ran a test expeirment over night (multiple stage
> positions and different z stacks) and it ran perfectly. Since then it
> has crashed about 15-20 times in the sapce of 3 days. We had a new super
> z stage installed at the same time as the upgrade (to correct some drift
> issues we were having). But i do not think the crashing is due to the
> super z stage because the software will crash when you use the zoom
> functions as well or use wide focus instead of galvo.
>
> Is anyone out there running version 1.8.0 of LAS AF and if so have you
> had similar issues. We have now downgraded the software back to 1.7.0
> and it seems to be running smoothly again. I just need to know if this
> is a global problem with ver 1.8.0 or it is something specific to our
> instrument. The leica technitian took 5 goes to get version 1.8
> installed as it kept failing at different parts of the firm ware upgrade
> process. Similarly when taking it back to ver 1.7 i ahd the same
> problems.
>
> On another note does anyone out there with a super z stage have it
> vibrate on them? Both stages we have had vibrate and emit a high pitched
> wine. If you poke it it will stop and everthing is fine. If it is
> vibrating when you are trying to capture an image then you don't get a
> good picture.
>
>
> Thanks
>
>
> Cam
>
>
> Cameron Nowell B. Sc (Hons)
> Microscopy Imaging and Research Core Facility
> Peter MacCallum Cancer Centre
> 7 St Andrews Place
> East Melbourne, Victoria 3002
> AUSTRALIA
> Phone: [phone]
> Mobile: [phone]
> Fax: [phone]
>
>
>
> This email (including any attachments) may contain
> confidential and/or legally privileged information and is
> intended only to be read or used by the addressee.  If you
> are not the intended addressee, any use, distribution,
> disclosure or copying of this email is strictly
> prohibited.
> Confidentiality and legal privilege attached to this email
> (including any attachments) are not waived or lost by
> reason of its mistaken delivery to you.
> If you have received this email in error, please delete it
> and notify us immediately by telephone or email.  Peter
> MacCallum Cancer Centre provides no guarantee that this
> transmission is free of virus or that it has not been
> intercepted or altered and will not be liable for any delay
> in its receipt.
>