Intensity changes with a Zeiss LSM 5 exciter

[POUVREAU SANDRINE]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi all.
We have a confocal microscope LSM 5 exciter (Zeiss) which is lately having
some troubles. In brief, the fluorescence intensity is changing during the
record, in a line –related pattern (in other word, the intensity changes happen
on one or several lines of the picture).
Either we have a good resolution/high intensity picture, and the intensity is
dropping, or we have a poor resolution/low intensity picture, and the intensity
is abruptly increasing. This seems to (although we are not sure yet about this
part) increase when the confocal is on for a long time.
Did somebody experience the same phenomenon with a similar system?
The technician from Zeiss told us it could be a problem with one the
acquisition cards. However, our end of the year budget does not allow us to
change them. Is there any way we can try to at least minimize the problem?
Thanks a lot.
Regards
Sandrine

[Mark Cannell]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Electronics components are more often all or none in failure modes.  
Connectors are a different issue tho'. and more likely to fail. Try re-
plugging in  all the connectors you can.  Also, I assume you have  
checked/tested and proved that it's not the laser illumination side of  
things?

Hope this helps

Mark


On 23/10/2010, at 8:19 AM, Sandrine Pouvreau wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all.
> We have a confocal microscope LSM 5 exciter (Zeiss) which is lately  
> having
> some troubles. In brief, the fluorescence intensity is changing  
> during the
> record, in a line –related pattern (in other word, the intensity  
> changes happen
> on one or several lines of the picture).
> Either we have a good resolution/high intensity picture, and the  
> intensity is
> dropping, or we have a poor resolution/low intensity picture, and  
> the intensity
> is abruptly increasing. This seems to (although we are not sure yet  
> about this
> part) increase when the confocal is on for a long time.
> Did somebody experience the same phenomenon with a similar system?
> The technician from Zeiss told us it could be a problem with one the
> acquisition cards. However, our end of the year budget does not  
> allow us to
> change them. Is there any way we can try to at least minimize the  
> problem?
> Thanks a lot.
> Regards
> Sandrine

[Stephen Cody-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Dear Sandrine,

I've seen this a couple of times with faulty pmt power supplies on Bio-Rad systems. As Mark suggests it could also be the laser. Do you see the same pattern when you collect images simultaneously with two detectors? If so probably the laser. Do you only see the problem with one laser or irrespective of which laser you use?

Good luck
Steve

Stephen H. Cody

On 23/10/2010, at 6:19 AM, Sandrine Pouvreau <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all.
> We have a confocal microscope LSM 5 exciter (Zeiss) which is lately having
> some troubles. In brief, the fluorescence intensity is changing during the
> record, in a line –related pattern (in other word, the intensity changes happen
> on one or several lines of the picture).
> Either we have a good resolution/high intensity picture, and the intensity is
> dropping, or we have a poor resolution/low intensity picture, and the intensity
> is abruptly increasing. This seems to (although we are not sure yet about this
> part) increase when the confocal is on for a long time.
> Did somebody experience the same phenomenon with a similar system?
> The technician from Zeiss told us it could be a problem with one the
> acquisition cards. However, our end of the year budget does not allow us to
> change them. Is there any way we can try to at least minimize the problem?
> Thanks a lot.
> Regards
> Sandrine

[POUVREAU SANDRINE]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi.
If I do a double wavelength record with line interleaving, one stimulated at 488 with the argon laser and collected with PMT1, and the other one stimulated at 543 with an helium neon laser and collected with PMT2, I see the problem occuring exactly at the same point on both pictures. Same thing if I use in addition a transmitted light detector. So I thought it could not be PMTs or laser, unless we have something affecting both lasers at the same time. Connectors is a good idea, I will try that one. Any other suggestion?
Thank you.
Sandrine

________________________________________
De : Confocal Microscopy List [[hidden email]] de la part de Stephen Cody [[hidden email]]
Date d'envoi : samedi 23 octobre 2010 05:29
À : [hidden email]
Objet : Re: Intensity changes with a Zeiss LSM 5 exciter

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Dear Sandrine,

I've seen this a couple of times with faulty pmt power supplies on Bio-Rad systems. As Mark suggests it could also be the laser. Do you see the same pattern when you collect images simultaneously with two detectors? If so probably the laser. Do you only see the problem with one laser or irrespective of which laser you use?

Good luck
Steve

Stephen H. Cody

On 23/10/2010, at 6:19 AM, Sandrine Pouvreau <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all.
> We have a confocal microscope LSM 5 exciter (Zeiss) which is lately having
> some troubles. In brief, the fluorescence intensity is changing during the
> record, in a line –related pattern (in other word, the intensity changes happen
> on one or several lines of the picture).
> Either we have a good resolution/high intensity picture, and the intensity is
> dropping, or we have a poor resolution/low intensity picture, and the intensity
> is abruptly increasing. This seems to (although we are not sure yet about this
> part) increase when the confocal is on for a long time.
> Did somebody experience the same phenomenon with a similar system?
> The technician from Zeiss told us it could be a problem with one the
> acquisition cards. However, our end of the year budget does not allow us to
> change them. Is there any way we can try to at least minimize the problem?
> Thanks a lot.
> Regards
> Sandrine

[Mark Cannell]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Interesting, check the AOM connections.

Cheers Mark


On 23/10/2010, at 11:33 PM, POUVREAU SANDRINE wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi.
> If I do a double wavelength record with line interleaving, one  
> stimulated at 488 with the argon laser and collected with PMT1, and  
> the other one stimulated at 543 with an helium neon laser and  
> collected with PMT2, I see the problem occuring exactly at the same  
> point on both pictures. Same thing if I use in addition a  
> transmitted light detector. So I thought it could not be PMTs or  
> laser, unless we have something affecting both lasers at the same  
> time. Connectors is a good idea, I will try that one. Any other  
> suggestion?
> Thank you.
> Sandrine
>
> ________________________________________
> De : Confocal Microscopy List [[hidden email]] de  
> la part de Stephen Cody [[hidden email]]
> Date d'envoi : samedi 23 octobre 2010 05:29
> À : [hidden email]
> Objet : Re: Intensity changes with a Zeiss LSM 5 exciter
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear Sandrine,
>
> I've seen this a couple of times with faulty pmt power supplies on  
> Bio-Rad systems. As Mark suggests it could also be the laser. Do you  
> see the same pattern when you collect images simultaneously with two  
> detectors? If so probably the laser. Do you only see the problem  
> with one laser or irrespective of which laser you use?
>
> Good luck
> Steve
>
> Stephen H. Cody
>
> On 23/10/2010, at 6:19 AM, Sandrine Pouvreau <[hidden email]
> > wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hi all.
>> We have a confocal microscope LSM 5 exciter (Zeiss) which is lately  
>> having
>> some troubles. In brief, the fluorescence intensity is changing  
>> during the
>> record, in a line –related pattern (in other word, the intensity  
>> changes happen
>> on one or several lines of the picture).
>> Either we have a good resolution/high intensity picture, and the  
>> intensity is
>> dropping, or we have a poor resolution/low intensity picture, and  
>> the intensity
>> is abruptly increasing. This seems to (although we are not sure yet  
>> about this
>> part) increase when the confocal is on for a long time.
>> Did somebody experience the same phenomenon with a similar system?
>> The technician from Zeiss told us it could be a problem with one the
>> acquisition cards. However, our end of the year budget does not  
>> allow us to
>> change them. Is there any way we can try to at least minimize the  
>> problem?
>> Thanks a lot.
>> Regards
>> Sandrine

[POUVREAU SANDRINE]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

You solved my problem! The AOM connections were indeed shaky.
Thanks a lot, you saved me a lot of headaches!
Kind regards
Sandrine




-----Message d'origine-----
De : Confocal Microscopy List [mailto:[hidden email]] De la part de Mark Cannell
Envoyé : samedi 23 octobre 2010 13:22
À : [hidden email]
Objet : Re: RE : Intensity changes with a Zeiss LSM 5 exciter

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Interesting, check the AOM connections.

Cheers Mark


On 23/10/2010, at 11:33 PM, POUVREAU SANDRINE wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi.
> If I do a double wavelength record with line interleaving, one  
> stimulated at 488 with the argon laser and collected with PMT1, and  
> the other one stimulated at 543 with an helium neon laser and  
> collected with PMT2, I see the problem occuring exactly at the same  
> point on both pictures. Same thing if I use in addition a  
> transmitted light detector. So I thought it could not be PMTs or  
> laser, unless we have something affecting both lasers at the same  
> time. Connectors is a good idea, I will try that one. Any other  
> suggestion?
> Thank you.
> Sandrine
>
> ________________________________________
> De : Confocal Microscopy List [[hidden email]] de  
> la part de Stephen Cody [[hidden email]]
> Date d'envoi : samedi 23 octobre 2010 05:29
> À : [hidden email]
> Objet : Re: Intensity changes with a Zeiss LSM 5 exciter
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear Sandrine,
>
> I've seen this a couple of times with faulty pmt power supplies on  
> Bio-Rad systems. As Mark suggests it could also be the laser. Do you  
> see the same pattern when you collect images simultaneously with two  
> detectors? If so probably the laser. Do you only see the problem  
> with one laser or irrespective of which laser you use?
>
> Good luck
> Steve
>
> Stephen H. Cody
>
> On 23/10/2010, at 6:19 AM, Sandrine Pouvreau <[hidden email]
> > wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hi all.
>> We have a confocal microscope LSM 5 exciter (Zeiss) which is lately  
>> having
>> some troubles. In brief, the fluorescence intensity is changing  
>> during the
>> record, in a line -related pattern (in other word, the intensity  
>> changes happen
>> on one or several lines of the picture).
>> Either we have a good resolution/high intensity picture, and the  
>> intensity is
>> dropping, or we have a poor resolution/low intensity picture, and  
>> the intensity
>> is abruptly increasing. This seems to (although we are not sure yet  
>> about this
>> part) increase when the confocal is on for a long time.
>> Did somebody experience the same phenomenon with a similar system?
>> The technician from Zeiss told us it could be a problem with one the
>> acquisition cards. However, our end of the year budget does not  
>> allow us to
>> change them. Is there any way we can try to at least minimize the  
>> problem?
>> Thanks a lot.
>> Regards
>> Sandrine