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laser alignment issue FluoView1000

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jerie

laser alignment issue FluoView1000

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Listers,

following the installation of a new air condition, we are not succeeding to
get any light after the scanhead of our Olympus FluoView1000 CLSM. End of
fiber, there is plenty of light. Behind the scanhead, no light arrives. We
work with an inverted IX 81 microscope.

Unfortunately the service situation in Brazil is very poor, so if there
were any suggestions how to proceed, I would rather try to check the
alignment myself.

Anyone having experience with the FluoView1000 scanhead?

Thank you, Jens

Dr. Jens Rietdorf, visiting scientist @ center for technological
development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
Brasil.

Straatman, Kees (Dr.)

Re: laser alignment issue FluoView1000

Dear Jens,

With behind the scanhead you mean no light at the objective or already at an earlier stage?

Kees

Dr Ir K.R. Straatman
Senior Experimental Officer
Advanced Imaging Facility
Centre for Core Biotechnology Services
University of Leicester
http://www2.le.ac.uk/colleges/medbiopsych/facilities-and-services/cbs/lite/aif

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of jens rietdorf
Sent: 04 April 2016 22:40
To: [hidden email]
Subject: laser alignment issue FluoView1000

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Listers,

following the installation of a new air condition, we are not succeeding to get any light after the scanhead of our Olympus FluoView1000 CLSM. End of fiber, there is plenty of light. Behind the scanhead, no light arrives. We work with an inverted IX 81 microscope.

Unfortunately the service situation in Brazil is very poor, so if there were any suggestions how to proceed, I would rather try to check the alignment myself.

Anyone having experience with the FluoView1000 scanhead?

Thank you, Jens

Dr. Jens Rietdorf, visiting scientist @ center for technological development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro Brasil.

jerie

Re: laser alignment issue FluoView1000

In reply to this post by jerie

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Listers,

according to suggestions from Olympus Americas and Avi Jacob (thanks!), I
have compared power measures with the internal light power meter (LPM) to
earlier measurements from August last year. All 6 lasers except the 405nm
line (39% but I noticed weaker signal and de-focus here before) show full
power (99-100%).

Both red LED's on the right side of the scan head are ON.
(filter turrets manual shutter open, filter block in mirror position1).

Still no light out of the scanhead.

All further suggestions welcome!

Very happy about the listserver!
Jens

Abraços,

Dr. Jens Rietdorf, visiting scientist @ center for technological
development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
Brasil.

On Mon, Apr 4, 2016 at 6:39 PM, jens rietdorf <[hidden email]> wrote:

> Listers,
>
> following the installation of a new air condition, we are not succeeding
> to get any light after the scanhead of our Olympus FluoView1000 CLSM. End
> of fiber, there is plenty of light. Behind the scanhead, no light arrives.
> We work with an inverted IX 81 microscope.
>
> Unfortunately the service situation in Brazil is very poor, so if there
> were any suggestions how to proceed, I would rather try to check the
> alignment myself.
>
> Anyone having experience with the FluoView1000 scanhead?
>
> Thank you, Jens
>
> Dr. Jens Rietdorf, visiting scientist @ center for technological
> development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
> Brasil.
>

jerie

Re: laser alignment issue FluoView1000

In reply to this post by Straatman, Kees (Dr.)

*****
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*****

I have removed the manual shutter from the filter turret, re-inserted the
filter turret and I am looking at a white paper inserted at the position of
the shutter. I have even looked directly onto the mirror cube to see if I
could see any residual light out of the scanhead with room lights switched
off: NO light whatsoever.

Zdenek Svindrych-2

Re: laser alignment issue FluoView1000

In reply to this post by jerie

*****
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*****

Hi,
in a normal point-scanning confocal there is no critical alignment in the
excitation path. I've seen come weird configurations (e.g. old Nikon PCM
2000), where both excitation light and the emission was directed through the
pinhole, there a small misalignment can result in complete loss of
excitation light. But I think FV1000 is not the case.

It must be either some safety shutter or the scanner is in parked position.
Can you hear the scanner?

Good luck!

zdenek

---------- Původní zpráva ----------
Od: jens rietdorf <[hidden email]>
Komu: [hidden email]
Datum: 5. 4. 2016 10:25:29
Předmět: Re: laser alignment issue FluoView1000

"*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Listers,

according to suggestions from Olympus Americas and Avi Jacob (thanks!), I
have compared power measures with the internal light power meter (LPM) to
earlier measurements from August last year. All 6 lasers except the 405nm
line (39% but I noticed weaker signal and de-focus here before) show full
power (99-100%).

Both red LED's on the right side of the scan head are ON.
(filter turrets manual shutter open, filter block in mirror position1).

Still no light out of the scanhead.

All further suggestions welcome!

Very happy about the listserver!
Jens

Abraços,

Dr. Jens Rietdorf, visiting scientist @ center for technological
development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
Brasil.

On Mon, Apr 4, 2016 at 6:39 PM, jens rietdorf <[hidden email]> wrote:

Janeiro
> Brasil.
>"

Smith, Benjamin E.

Re: laser alignment issue FluoView1000

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

I'm not sure where the AOTF is in the FV1000 light path, but when an AOTF dies, the laser output drops nearly to zero (although an extremely faint beam is still transmitted). This may also be something to look into. However, the fact that both the UV and visible lines kicked the bucket, it seems unlikely to me that both AOTFs would die at the same time.

Another problem we've had on older systems is the filter turret/slider stops indexing properly, leading to little or no light depending on the filter position. Manually pushing the turret to a roughly centered position, and then letting the scan head reinitialize, correctly reset the indexing.

Along these lines, you should be able to see and hear each mechanical component in the scan head initialize. A failure of a mechanical component to correctly find its zero position (i.e. jitters side to side, moves to the extreme of the device's range, or doesn't move at all) is a quick way to determine if you may have a mechanical failure. What can sometimes happen is that a component rotated past it's expected range (effectively a negative position). As a result, if the system wasn't designed to understand a negative position, it will try and initialize the part by moving it further in the negative direction, leaving it in some extreme position (look up "integer overflow" for more info on this: https://en.wikipedia.org/wiki/Integer_overflow).

This is why recentering the part can sometimes help, because the system can then correctly interpret the part's position and then zero it correctly when the scanhead is reinitialized. I found this problem tends to be especially predominant on older systems.

Good luck,
Ben Smith

________________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Zdenek Svindrych <[hidden email]>
Sent: Tuesday, April 5, 2016 9:38 AM
To: [hidden email]
Subject: Re: laser alignment issue FluoView1000

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi,
in a normal point-scanning confocal there is no critical alignment in the
excitation path. I've seen come weird configurations (e.g. old Nikon PCM
2000), where both excitation light and the emission was directed through the
pinhole, there a small misalignment can result in complete loss of
excitation light. But I think FV1000 is not the case.

It must be either some safety shutter or the scanner is in parked position.
Can you hear the scanner?

Good luck!

zdenek

---------- Původní zpráva ----------
Od: jens rietdorf <[hidden email]>
Komu: [hidden email]
Datum: 5. 4. 2016 10:25:29
Předmět: Re: laser alignment issue FluoView1000

"*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Listers,

according to suggestions from Olympus Americas and Avi Jacob (thanks!), I
have compared power measures with the internal light power meter (LPM) to
earlier measurements from August last year. All 6 lasers except the 405nm
line (39% but I noticed weaker signal and de-focus here before) show full
power (99-100%).

Both red LED's on the right side of the scan head are ON.
(filter turrets manual shutter open, filter block in mirror position1).

Still no light out of the scanhead.

All further suggestions welcome!

Very happy about the listserver!
Jens

Abraços,

Dr. Jens Rietdorf, visiting scientist @ center for technological
development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
Brasil.

On Mon, Apr 4, 2016 at 6:39 PM, jens rietdorf <[hidden email]> wrote:

Janeiro
> Brasil.
>"

Smith, Benjamin E.

Re: laser alignment issue FluoView1000

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

One other thing that just came to mind that we see a bit on our FV500 (and could have been directly caused by the install) is that if the data cables going to the scan head or computer get pulled or jostled, then the scan head more or less stops working. This is often (but not always) indicated by the software failing to boot up the scan head altogether. That said, we have had one time where the scan head did boot, and the software allowed us to control the scan head. However, while we could get the arc lamp to shine out the objective, we could not get an image in laser scanning mode.

Almost every problem we've had with the FV500 has been fixed by replacing the problematic data cable. As such, I've now fastened the cables to fixed structures so that there is no tension in the system, and we have not seen this problem since (knock on wood).

-Ben Smith

________________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Smith, Benjamin E. <[hidden email]>
Sent: Tuesday, April 5, 2016 12:08 PM
To: [hidden email]
Subject: Re: laser alignment issue FluoView1000

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

I'm not sure where the AOTF is in the FV1000 light path, but when an AOTF dies, the laser output drops nearly to zero (although an extremely faint beam is still transmitted). This may also be something to look into. However, the fact that both the UV and visible lines kicked the bucket, it seems unlikely to me that both AOTFs would die at the same time.

Another problem we've had on older systems is the filter turret/slider stops indexing properly, leading to little or no light depending on the filter position. Manually pushing the turret to a roughly centered position, and then letting the scan head reinitialize, correctly reset the indexing.

Along these lines, you should be able to see and hear each mechanical component in the scan head initialize. A failure of a mechanical component to correctly find its zero position (i.e. jitters side to side, moves to the extreme of the device's range, or doesn't move at all) is a quick way to determine if you may have a mechanical failure. What can sometimes happen is that a component rotated past it's expected range (effectively a negative position). As a result, if the system wasn't designed to understand a negative position, it will try and initialize the part by moving it further in the negative direction, leaving it in some extreme position (look up "integer overflow" for more info on this: https://en.wikipedia.org/wiki/Integer_overflow).

This is why recentering the part can sometimes help, because the system can then correctly interpret the part's position and then zero it correctly when the scanhead is reinitialized. I found this problem tends to be especially predominant on older systems.

Good luck,
Ben Smith

________________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Zdenek Svindrych <[hidden email]>
Sent: Tuesday, April 5, 2016 9:38 AM
To: [hidden email]
Subject: Re: laser alignment issue FluoView1000

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi,
in a normal point-scanning confocal there is no critical alignment in the
excitation path. I've seen come weird configurations (e.g. old Nikon PCM
2000), where both excitation light and the emission was directed through the
pinhole, there a small misalignment can result in complete loss of
excitation light. But I think FV1000 is not the case.

It must be either some safety shutter or the scanner is in parked position.
Can you hear the scanner?

Good luck!

zdenek

---------- Původní zpráva ----------
Od: jens rietdorf <[hidden email]>
Komu: [hidden email]
Datum: 5. 4. 2016 10:25:29
Předmět: Re: laser alignment issue FluoView1000

"*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Listers,

according to suggestions from Olympus Americas and Avi Jacob (thanks!), I
have compared power measures with the internal light power meter (LPM) to
earlier measurements from August last year. All 6 lasers except the 405nm
line (39% but I noticed weaker signal and de-focus here before) show full
power (99-100%).

Both red LED's on the right side of the scan head are ON.
(filter turrets manual shutter open, filter block in mirror position1).

Still no light out of the scanhead.

All further suggestions welcome!

Very happy about the listserver!
Jens

Abraços,

Dr. Jens Rietdorf, visiting scientist @ center for technological
development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
Brasil.

On Mon, Apr 4, 2016 at 6:39 PM, jens rietdorf <[hidden email]> wrote:

Janeiro
> Brasil.
>"

Glen MacDonald-2

Re: laser alignment issue FluoView1000

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Shortly after our FV-1000 was installed, I bolted a pillar to the table, in front of the cable ports on the right side of the IX-8. that eliminated the many calls from users who could not get a scan or the software would not full load. They were knocking cables loose by setting their notebooks or slide boxes on the table then shoving them back out of the way. It is just a 6 inch long scrap of Thorlabs 25 mm extruded aluminum.

Good luck,
Glen MacDonald
Digital Microscopy Center
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
[hidden email]

> On Apr 5, 2016, at 10:20 AM, Smith, Benjamin E. <[hidden email]> wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> One other thing that just came to mind that we see a bit on our FV500 (and could have been directly caused by the install) is that if the data cables going to the scan head or computer get pulled or jostled, then the scan head more or less stops working. This is often (but not always) indicated by the software failing to boot up the scan head altogether. That said, we have had one time where the scan head did boot, and the software allowed us to control the scan head. However, while we could get the arc lamp to shine out the objective, we could not get an image in laser scanning mode.
>
> Almost every problem we've had with the FV500 has been fixed by replacing the problematic data cable. As such, I've now fastened the cables to fixed structures so that there is no tension in the system, and we have not seen this problem since (knock on wood).
>
> -Ben Smith
>
> ________________________________________
> From: Confocal Microscopy List <[hidden email]> on behalf of Smith, Benjamin E. <[hidden email]>
> Sent: Tuesday, April 5, 2016 12:08 PM
> To: [hidden email]
> Subject: Re: laser alignment issue FluoView1000
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> I'm not sure where the AOTF is in the FV1000 light path, but when an AOTF dies, the laser output drops nearly to zero (although an extremely faint beam is still transmitted). This may also be something to look into. However, the fact that both the UV and visible lines kicked the bucket, it seems unlikely to me that both AOTFs would die at the same time.
>
> Another problem we've had on older systems is the filter turret/slider stops indexing properly, leading to little or no light depending on the filter position. Manually pushing the turret to a roughly centered position, and then letting the scan head reinitialize, correctly reset the indexing.
>
> Along these lines, you should be able to see and hear each mechanical component in the scan head initialize. A failure of a mechanical component to correctly find its zero position (i.e. jitters side to side, moves to the extreme of the device's range, or doesn't move at all) is a quick way to determine if you may have a mechanical failure. What can sometimes happen is that a component rotated past it's expected range (effectively a negative position). As a result, if the system wasn't designed to understand a negative position, it will try and initialize the part by moving it further in the negative direction, leaving it in some extreme position (look up "integer overflow" for more info on this: https://en.wikipedia.org/wiki/Integer_overflow).
>
> This is why recentering the part can sometimes help, because the system can then correctly interpret the part's position and then zero it correctly when the scanhead is reinitialized. I found this problem tends to be especially predominant on older systems.
>
> Good luck,
> Ben Smith
>
> ________________________________________
> From: Confocal Microscopy List <[hidden email]> on behalf of Zdenek Svindrych <[hidden email]>
> Sent: Tuesday, April 5, 2016 9:38 AM
> To: [hidden email]
> Subject: Re: laser alignment issue FluoView1000
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi,
> in a normal point-scanning confocal there is no critical alignment in the
> excitation path. I've seen come weird configurations (e.g. old Nikon PCM
> 2000), where both excitation light and the emission was directed through the
> pinhole, there a small misalignment can result in complete loss of
> excitation light. But I think FV1000 is not the case.
>
> It must be either some safety shutter or the scanner is in parked position.
> Can you hear the scanner?
>
> Good luck!
>
> zdenek
>
>
> ---------- Původní zpráva ----------
> Od: jens rietdorf <[hidden email]>
> Komu: [hidden email]
> Datum: 5. 4. 2016 10:25:29
> Předmět: Re: laser alignment issue FluoView1000
>
> "*****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Listers,
>
> according to suggestions from Olympus Americas and Avi Jacob (thanks!), I
> have compared power measures with the internal light power meter (LPM) to
> earlier measurements from August last year. All 6 lasers except the 405nm
> line (39% but I noticed weaker signal and de-focus here before) show full
> power (99-100%).
>
> Both red LED's on the right side of the scan head are ON.
> (filter turrets manual shutter open, filter block in mirror position1).
>
> Still no light out of the scanhead.
>
> All further suggestions welcome!
>
> Very happy about the listserver!
> Jens
>
> Abraços,
>
> Dr. Jens Rietdorf, visiting scientist @ center for technological
> development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
> Brasil.
>
> On Mon, Apr 4, 2016 at 6:39 PM, jens rietdorf <[hidden email]> wrote:
>
>> Listers,
>>
>> following the installation of a new air condition, we are not succeeding
>> to get any light after the scanhead of our Olympus FluoView1000 CLSM. End
>> of fiber, there is plenty of light. Behind the scanhead, no light arrives.
>> We work with an inverted IX 81 microscope.
>>
>> Unfortunately the service situation in Brazil is very poor, so if there
>> were any suggestions how to proceed, I would rather try to check the
>> alignment myself.
>>
>> Anyone having experience with the FluoView1000 scanhead?
>>
>> Thank you, Jens
>>
>> Dr. Jens Rietdorf, visiting scientist @ center for technological
>> development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de
> Janeiro
>> Brasil.
>> "

Jacqueline Ross

Re: laser alignment issue FluoView1000

In reply to this post by jerie

Hi Jens,

To me, it doesn't sound like alignment per se as you don't have any light from any lasers.

Did you have to dismantle the confocal in order to have the A/C installed. If so, there might be a polariser somewhere in the scan head that's affecting things. We had a laser replaced and the engineer couldn't get any light out of the lens initially though everything was fine on the fibre side. Then it turned out that there was a polariser in the system (don't know where sorry:-() that was blocking all of the light!

The other thing I thought I'd mention (though am a bit embarrassed to do so) is that we often have people turning off the keys on our scanner power supplies. There's always a bit of puzzlement that the system won't scan since the indicator light does come on with the switch even when the keys are in the OFF position.

Checking the cables between the scanner power supplies and scan head as has been suggested is a very good idea too though we've never had any problems with that as ours are all at the back.

Kind regards,

Jacqui

Jacqueline Ross
Biomedical Imaging Microscopist
Biomedical Imaging Research Unit
School of Medical Sciences
Faculty of Medical & Health Sciences
The University of Auckland
Private Bag 92019
Auckland 1142, NEW ZEALAND

Tel: 64 9 923 7438
Fax: 64 9 373 7484

http://www.fmhs.auckland.ac.nz/sms/biru/

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of jens rietdorf
Sent: Tuesday, 5 April 2016 9:40 a.m.
To: [hidden email]
Subject: laser alignment issue FluoView1000

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Listers,

following the installation of a new air condition, we are not succeeding to
get any light after the scanhead of our Olympus FluoView1000 CLSM. End of
fiber, there is plenty of light. Behind the scanhead, no light arrives. We
work with an inverted IX 81 microscope.

Unfortunately the service situation in Brazil is very poor, so if there
were any suggestions how to proceed, I would rather try to check the
alignment myself.

Anyone having experience with the FluoView1000 scanhead?

Thank you, Jens

Dr. Jens Rietdorf, visiting scientist @ center for technological
development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
Brasil.

jerie

Re: laser alignment issue FluoView1000

In reply to this post by jerie

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear colleagues,

thank you all for your help on the issue, it turned out a shutter inside
the scanhead was blocked. This problem was relatively rapidly addressed by
the service of Olympus Brasil and Olympus Americas. Very helpful input for
the differential diagnosis came from Farid Jalali of Olympus Canada.

A new (maybe related) issue occurred now, we experience very low
sensitivity on detector channels 1&2,
channel 3 operates normally. the effect appears to be stronger in the
blue/green part of the spectrum than for the red.
The FV diagnostic tool does not indicate any abnormalities.

Again, all ideas and suggestions are very welcome.

Kind regards, Jens

Dr. Jens Rietdorf, visiting scientist @ center for technological
development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
Brasil.

On Mon, Apr 4, 2016 at 6:39 PM, jens rietdorf <[hidden email]> wrote:

Glen MacDonald-2

Re: laser alignment issue FluoView1000

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Jens,
Have you tried redirecting the blue and green channels to the detector that is usually “red”?
after ~28,000 hrs on our FV-1000, the “green” channel started dropping out, sometimes entirely, other times with dark bands in the scanned frame. The problem persisted when routed to other detectors. The culprit was the AOTF.

Good luck,
Glen
Glen MacDonald
Digital Microscopy Center
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
[hidden email]

> On Jun 1, 2016, at 10:35 AM, jens rietdorf <[hidden email]> wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear colleagues,
>
> thank you all for your help on the issue, it turned out a shutter inside
> the scanhead was blocked. This problem was relatively rapidly addressed by
> the service of Olympus Brasil and Olympus Americas. Very helpful input for
> the differential diagnosis came from Farid Jalali of Olympus Canada.
>
> A new (maybe related) issue occurred now, we experience very low
> sensitivity on detector channels 1&2,
> channel 3 operates normally. the effect appears to be stronger in the
> blue/green part of the spectrum than for the red.
> The FV diagnostic tool does not indicate any abnormalities.
>
> Again, all ideas and suggestions are very welcome.
>
> Kind regards, Jens
>
> Dr. Jens Rietdorf, visiting scientist @ center for technological
> development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
> Brasil.
>
> On Mon, Apr 4, 2016 at 6:39 PM, jens rietdorf <[hidden email]> wrote:
>
>> Listers,
>>
>> following the installation of a new air condition, we are not succeeding
>> to get any light after the scanhead of our Olympus FluoView1000 CLSM. End
>> of fiber, there is plenty of light. Behind the scanhead, no light arrives.
>> We work with an inverted IX 81 microscope.
>>
>> Unfortunately the service situation in Brazil is very poor, so if there
>> were any suggestions how to proceed, I would rather try to check the
>> alignment myself.
>>
>> Anyone having experience with the FluoView1000 scanhead?
>>
>> Thank you, Jens
>>
>> Dr. Jens Rietdorf, visiting scientist @ center for technological
>> development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
>> Brasil.
>>

jerie

Re: laser alignment issue FluoView1000

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Hi Glen,

exactly that was what I did to find out detector 1&2 are far weaker than
ch3. A good guess seems to be 'Spectral Detectors out of tune' which was
the remote diagnosis by Olympus that just arrived.

Anyone with experience in tuning Spectral Detectors on the FV1000?

Thanks a lot,
jens

Abraços,

Dr. Jens Rietdorf, visiting scientist @ center for technological
development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de Janeiro
Brasil.

On Wed, Jun 1, 2016 at 3:34 PM, Glen MacDonald <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hi Jens,
> Have you tried redirecting the blue and green channels to the detector
> that is usually “red”?
> after ~28,000 hrs on our FV-1000, the “green” channel started dropping
> out, sometimes entirely, other times with dark bands in the scanned frame.
> The problem persisted when routed to other detectors. The culprit was the
> AOTF.
>
> Good luck,
> Glen
> Glen MacDonald
> Digital Microscopy Center
> Box 357923
> University of Washington
> Seattle, WA 98195-7923 USA
> (206) 616-4156
> [hidden email]
>
>
>
>
>
>
>
>
> > On Jun 1, 2016, at 10:35 AM, jens rietdorf <[hidden email]> wrote:
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your
> posting.
> > *****
> >
> > Dear colleagues,
> >
> > thank you all for your help on the issue, it turned out a shutter inside
> > the scanhead was blocked. This problem was relatively rapidly addressed
> by
> > the service of Olympus Brasil and Olympus Americas. Very helpful input
> for
> > the differential diagnosis came from Farid Jalali of Olympus Canada.
> >
> > A new (maybe related) issue occurred now, we experience very low
> > sensitivity on detector channels 1&2,
> > channel 3 operates normally. the effect appears to be stronger in the
> > blue/green part of the spectrum than for the red.
> > The FV diagnostic tool does not indicate any abnormalities.
> >
> > Again, all ideas and suggestions are very welcome.
> >
> > Kind regards, Jens
> >
> > Dr. Jens Rietdorf, visiting scientist @ center for technological
> > development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de
> Janeiro
> > Brasil.
> >
> > On Mon, Apr 4, 2016 at 6:39 PM, jens rietdorf <[hidden email]>
> wrote:
> >
> >> Listers,
> >>
> >> following the installation of a new air condition, we are not succeeding
> >> to get any light after the scanhead of our Olympus FluoView1000 CLSM.
> End
> >> of fiber, there is plenty of light. Behind the scanhead, no light
> arrives.
> >> We work with an inverted IX 81 microscope.
> >>
> >> Unfortunately the service situation in Brazil is very poor, so if there
> >> were any suggestions how to proceed, I would rather try to check the
> >> alignment myself.
> >>
> >> Anyone having experience with the FluoView1000 scanhead?
> >>
> >> Thank you, Jens
> >>
> >> Dr. Jens Rietdorf, visiting scientist @ center for technological
> >> development in health CDTS, Oswaldo Cruz Foundation Fiocruz, Rio de
> Janeiro
> >> Brasil.
> >>
>

Stanislav Vitha-2

Re: laser alignment issue FluoView1000

In reply to this post by jerie

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Sometimes, the motorized slit in the spectral detector unit gets stuck, so if
it is stuck in a narrow bandwidth setting, you would be getting very little
signal. But it is unlikely that this would happen on both detectors the same
time.
You could try changing the bandwidth from the maximum 100 nm down 10
nm when imaging a fluorescent slide or a solution of a fluorescent dye. If
the slit adjustment works, the signal will go down.

Another possibility is that the spectral calibration is off, but again not likely
to happen for both detector units, unless somebody played with the
settings, logged as Maker (administrator). One easy way to test is to image
a surface in reflected mode, with minimum spectral bandwidth (2nm). For
instance, use 488 nm laser, set the detector to 488-490nm detection. In
live mode, start moving the detector band in 1 nm steps up or down and
watch the signal intensity. Once you get out of the laser wavelength range,
the signal should drop very fast.

Stan

Dr. Stanislav Vitha
Texas A&M University
Microscopy and Imaging Center