odd inensity shift over time
Locked
|
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear confocalers, We would appreciate help identifying (and remedying) a problem we have not encountered before. One of our users reported that the laser intensity of the confocal was dropping off during each of her imaging sessions. Today I put a mirror on the confocal and imaged for 4 1/2 hours in the reflectance mode with the slits to detect over than 20 nm open. During that time the intensities at 488, 543, and 633 nm rose to a peak between 15 and 60 minutes and then fell off at a slower rate (graph posted at http://www.flickr.com/photos/mcammer/6512342071/ ). Has anyone seen this pattern before and could help us identify the cause? Is it a laser issue (with three different lasers?), an AOTF issue, a detectors issue, a drifting out of alignment issue (although in this last case the resolution in fluorescent imaging continues to look fine)? Also, this is not a focus drift issue. The microscope is a Zeiss 710 with three spectral detectors. Any help identifying this greatly appreciated. ________________________________________________________ Michael Cammer, Assistant Research Scientist Skirball Institute of Biomolecular Medicine Lab: (212) 263-3208 Cell: (914) 309-3270 </PRE> <html> <body> ------------------------------------------------------------<br /> This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email.<br /> ================================= </body> </html> <PRE> |
 
|
Craig Brideau |
Re: odd inensity shift over time
|
|
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Is the temperature in the room holding steady over this time period? Craig On Wed, Dec 14, 2011 at 1:44 PM, Cammer, Michael <[hidden email] > wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear confocalers, > We would appreciate help identifying (and remedying) a problem we have not > encountered before. > One of our users reported that the laser intensity of the confocal was > dropping off during each of her imaging sessions. Today I put a mirror on > the confocal and imaged for 4 1/2 hours in the reflectance mode with the > slits to detect over than 20 nm open. During that time the intensities at > 488, 543, and 633 nm rose to a peak between 15 and 60 minutes and then fell > off at a slower rate (graph posted at > http://www.flickr.com/photos/mcammer/6512342071/ ). > Has anyone seen this pattern before and could help us identify the cause? > Is it a laser issue (with three different lasers?), an AOTF issue, a > detectors issue, a drifting out of alignment issue (although in this last > case the resolution in fluorescent imaging continues to look fine)? > Also, this is not a focus drift issue. > > The microscope is a Zeiss 710 with three spectral detectors. > > Any help identifying this greatly appreciated. > ________________________________________________________ > Michael Cammer, Assistant Research Scientist > Skirball Institute of Biomolecular Medicine > Lab: (212) 263-3208 Cell: (914) 309-3270 > > </PRE> > <html> > <body> > ------------------------------------------------------------<br /> > This email message, including any attachments, is for the sole use of the > intended recipient(s) and may contain information that is proprietary, > confidential, and exempt from disclosure under applicable law. Any > unauthorized review, use, disclosure, or distribution is prohibited. If you > have received this email in error please notify the sender by return email > and delete the original message. Please note, the recipient should check > this email and any attachments for the presence of viruses. The > organization accepts no liability for any damage caused by any virus > transmitted by this email.<br /> > ================================= > </body> > </html> > <PRE> > |
|
|
Vladimir Ghukasyan-2 |
Re: odd inensity shift over time
|
|
In reply to this post by mcammer
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Michael, Does this happen when you run imaging with a single laser at a time? Vladimir On Wed, Dec 14, 2011 at 3:44 PM, Cammer, Michael <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear confocalers, > We would appreciate help identifying (and remedying) a problem we have not encountered before. > One of our users reported that the laser intensity of the confocal was dropping off during each of her imaging sessions. Today I put a mirror on the confocal and imaged for 4 1/2 hours in the reflectance mode with the slits to detect over than 20 nm open. During that time the intensities at 488, 543, and 633 nm rose to a peak between 15 and 60 minutes and then fell off at a slower rate (graph posted at http://www.flickr.com/photos/mcammer/6512342071/ ). > Has anyone seen this pattern before and could help us identify the cause? Is it a laser issue (with three different lasers?), an AOTF issue, a detectors issue, a drifting out of alignment issue (although in this last case the resolution in fluorescent imaging continues to look fine)? > Also, this is not a focus drift issue. > > The microscope is a Zeiss 710 with three spectral detectors. > > Any help identifying this greatly appreciated. > ________________________________________________________ > Michael Cammer, Assistant Research Scientist > Skirball Institute of Biomolecular Medicine > Lab: (212) 263-3208 Cell: (914) 309-3270 > > </PRE> > <html> > <body> > ------------------------------------------------------------<br /> > This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email.<br /> > ================================= > </body> > </html> > <PRE> |
|
|
In reply to this post by mcammer
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear Michael, I could imagine that it I the temperature stabilization/control of the AOTF that creates the observed phenomenon. You could relatively easily test this if you would do an intensity measurement directly on the laser (before the AOTF) and then do the same thing after the AOTF (but before the fibre). Cheers Gabor On 12/14/11 9:44 PM, "Cammer, Michael" <[hidden email]> wrote: >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >Dear confocalers, >We would appreciate help identifying (and remedying) a problem we have >not encountered before. >One of our users reported that the laser intensity of the confocal was >dropping off during each of her imaging sessions. Today I put a mirror >on the confocal and imaged for 4 1/2 hours in the reflectance mode with >the slits to detect over than 20 nm open. During that time the >intensities at 488, 543, and 633 nm rose to a peak between 15 and 60 >minutes and then fell off at a slower rate (graph posted at >http://www.flickr.com/photos/mcammer/6512342071/ ). >Has anyone seen this pattern before and could help us identify the cause? > Is it a laser issue (with three different lasers?), an AOTF issue, a >detectors issue, a drifting out of alignment issue (although in this last >case the resolution in fluorescent imaging continues to look fine)? >Also, this is not a focus drift issue. > >The microscope is a Zeiss 710 with three spectral detectors. > >Any help identifying this greatly appreciated. >________________________________________________________ >Michael Cammer, Assistant Research Scientist >Skirball Institute of Biomolecular Medicine >Lab: (212) 263-3208 Cell: (914) 309-3270 > ></PRE> ><html> ><body> >------------------------------------------------------------<br /> >This email message, including any attachments, is for the sole use of the >intended recipient(s) and may contain information that is proprietary, >confidential, and exempt from disclosure under applicable law. Any >unauthorized review, use, disclosure, or distribution is prohibited. If >you have received this email in error please notify the sender by return >email and delete the original message. Please note, the recipient should >check this email and any attachments for the presence of viruses. The >organization accepts no liability for any damage caused by any virus >transmitted by this email.<br /> >================================= ></body> ></html> ><PRE> |
|
|
In reply to this post by mcammer
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I would first check what's going on in the transmitted detector. Mike -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Cammer, Michael Sent: Wednesday, December 14, 2011 3:44 PM To: [hidden email] Subject: odd inensity shift over time ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear confocalers, We would appreciate help identifying (and remedying) a problem we have not encountered before. One of our users reported that the laser intensity of the confocal was dropping off during each of her imaging sessions. Today I put a mirror on the confocal and imaged for 4 1/2 hours in the reflectance mode with the slits to detect over than 20 nm open. During that time the intensities at 488, 543, and 633 nm rose to a peak between 15 and 60 minutes and then fell off at a slower rate (graph posted at http://www.flickr.com/photos/mcammer/6512342071/ ). Has anyone seen this pattern before and could help us identify the cause? Is it a laser issue (with three different lasers?), an AOTF issue, a detectors issue, a drifting out of alignment issue (although in this last case the resolution in fluorescent imaging continues to look fine)? Also, this is not a focus drift issue. The microscope is a Zeiss 710 with three spectral detectors. Any help identifying this greatly appreciated. ________________________________________________________ Michael Cammer, Assistant Research Scientist Skirball Institute of Biomolecular Medicine Lab: (212) 263-3208 Cell: (914) 309-3270 </PRE> <html> <body> ------------------------------------------------------------<br /> This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email.<br /> ================================= </body> </html> <PRE> |
|
|
Boswell, Carl A - (cboswell) |
Re: odd inensity shift over time
|
|
My first guess would be the AOTF, notwithstanding the differences in spectral changes. Does Zeiss have any comment?
C Carl A. Boswell 520-954-7053 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of MODEL, MICHAEL Sent: Wednesday, December 14, 2011 2:25 PM To: [hidden email] Subject: Re: odd inensity shift over time ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I would first check what's going on in the transmitted detector. Mike -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Cammer, Michael Sent: Wednesday, December 14, 2011 3:44 PM To: [hidden email] Subject: odd inensity shift over time ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear confocalers, We would appreciate help identifying (and remedying) a problem we have not encountered before. One of our users reported that the laser intensity of the confocal was dropping off during each of her imaging sessions. Today I put a mirror on the confocal and imaged for 4 1/2 hours in the reflectance mode with the slits to detect over than 20 nm open. During that time the intensities at 488, 543, and 633 nm rose to a peak between 15 and 60 minutes and then fell off at a slower rate (graph posted at http://www.flickr.com/photos/mcammer/6512342071/ ). Has anyone seen this pattern before and could help us identify the cause? Is it a laser issue (with three different lasers?), an AOTF issue, a detectors issue, a drifting out of alignment issue (although in this last case the resolution in fluorescent imaging continues to look fine)? Also, this is not a focus drift issue. The microscope is a Zeiss 710 with three spectral detectors. Any help identifying this greatly appreciated. ________________________________________________________ Michael Cammer, Assistant Research Scientist Skirball Institute of Biomolecular Medicine Lab: (212) 263-3208 Cell: (914) 309-3270 </PRE> <html> <body> ------------------------------------------------------------<br /> This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email.<br /> ================================= </body> </html> <PRE> |
|
|
Zac Arrac Atelaz |
Re: odd inensity shift over time
|
|
In reply to this post by mcammer
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Michael: Some things come to my mind, not sure they are what you are looking for, but use them as reference: 1. Change on the ambient temperature, It would be great having an adjacent graph with the room and laser area temp on side of this one, gas laser is making a blinking, fast enough so you cannot notice, changes in temp would make this faster or slower, and also affect the efficiency of the light emitting properties of the device. 2. If you want to check, there is a laser stabilization function which is basically a feedback info going to the AOTF, I am sure Olympus have this on the FV1000, but you have to check if Zeiss can retrofit something like this to your system. gas laser is on a continuos fluctuation, which might be corrected with this feedback function. Hope this helps Gabriel OH > Date: Wed, 14 Dec 2011 15:44:15 -0500 > From: [hidden email] > Subject: odd inensity shift over time > To: [hidden email] > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear confocalers, > We would appreciate help identifying (and remedying) a problem we have not encountered before. > One of our users reported that the laser intensity of the confocal was dropping off during each of her imaging sessions. Today I put a mirror on the confocal and imaged for 4 1/2 hours in the reflectance mode with the slits to detect over than 20 nm open. During that time the intensities at 488, 543, and 633 nm rose to a peak between 15 and 60 minutes and then fell off at a slower rate (graph posted at http://www.flickr.com/photos/mcammer/6512342071/ ). > Has anyone seen this pattern before and could help us identify the cause? Is it a laser issue (with three different lasers?), an AOTF issue, a detectors issue, a drifting out of alignment issue (although in this last case the resolution in fluorescent imaging continues to look fine)? > Also, this is not a focus drift issue. > > The microscope is a Zeiss 710 with three spectral detectors. > > Any help identifying this greatly appreciated. > ________________________________________________________ > Michael Cammer, Assistant Research Scientist > Skirball Institute of Biomolecular Medicine > Lab: (212) 263-3208 Cell: (914) 309-3270 > > </PRE> > <html> > <body> > ------------------------------------------------------------<br /> > This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email.<br /> > ================================= > </body> > </html> > <PRE> |
|
|
James Pawley |
Re: odd inensity shift over time
|
|
In reply to this post by mmodel
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Or better yet, just monitor the light coming out of a (low NA) objective with a simple photometer. Motion of the laser-end of the fibre with respect to the "objective" that focuses the laser light into it, might have times constants of the length you mention. JP *************************************************************************** Prof. James B. Pawley, Ph. 608-238-3953 21. N. Prospect Ave. Madison, WI 53726 USA [hidden email] 3D Microscopy of Living Cells Course, June 9-21, 2012, UBC, Vancouver Canada Info: http://www.3dcourse.ubc.ca/ Application deadline 3/16/2012 "If it ain't diffraction, it must be statistics." Anon. >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >I would first check what's going on in the transmitted detector. >Mike > >-----Original Message----- >From: Confocal Microscopy List >[mailto:[hidden email]] On Behalf Of Cammer, >Michael >Sent: Wednesday, December 14, 2011 3:44 PM >To: [hidden email] >Subject: odd inensity shift over time > >***** >To join, leave or search the confocal microscopy listserv, go to: >http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy >***** > >Dear confocalers, >We would appreciate help identifying (and remedying) a problem we >have not encountered before. >One of our users reported that the laser intensity of the confocal >was dropping off during each of her imaging sessions. Today I put >a mirror on the confocal and imaged for 4 1/2 hours in the >reflectance mode with the slits to detect over than 20 nm open. >During that time the intensities at 488, 543, and 633 nm rose to a >peak between 15 and 60 minutes and then fell off at a slower rate >(graph posted at http://www.flickr.com/photos/mcammer/6512342071/ ). >Has anyone seen this pattern before and could help us identify the >cause? Is it a laser issue (with three different lasers?), an AOTF >issue, a detectors issue, a drifting out of alignment issue >(although in this last case the resolution in fluorescent imaging >continues to look fine)? >Also, this is not a focus drift issue. > >The microscope is a Zeiss 710 with three spectral detectors. > >Any help identifying this greatly appreciated. >________________________________________________________ >Michael Cammer, Assistant Research Scientist >Skirball Institute of Biomolecular Medicine >Lab: (212) 263-3208 Cell: (914) 309-3270 > ></PRE> ><html> ><body> >------------------------------------------------------------<br /> >This email message, including any attachments, is for the sole use >of the intended recipient(s) and may contain information that is >proprietary, confidential, and exempt from disclosure under >applicable law. Any unauthorized review, use, disclosure, or >distribution is prohibited. If you have received this email in error >please notify the sender by return email and delete the original >message. Please note, the recipient should check this email and any >attachments for the presence of viruses. The organization accepts no >liability for any damage caused by any virus transmitted by this >email.<br /> >================================= ></body> ></html> ><PRE> -- |
|
|
Zucker.Robert |
Re: odd intensity shift over time
|
|
In reply to this post by mcammer
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear Mike This pattern probably has to do with the AOTF instability. We have published a few papers on the instability of a confocal microscope that are available on request. The data is quite disturbing, but it is believed that this effect can occur with AOTFs that are not heat stabilized or are defective. Please contact me directly for PDF copies of these publications best wishes bob 1. Zucker RM Confocal Slide Based System :Performance. Cytometry 2006 69 A 659-676.. 2. Zucker RM Confocal Microscopy Slide Based Systems: Instability. Cytometry 2006 69A 677-690. 3. Zucker, R.M. Evaluation of Confocal Microscopy System Performance. Cell Imaging Techniques. Douglas Taajets editor Humana Press Chapter 5 77-135 2005 Dr. Robert Zucker, PhD USEPA ORD NHEERL TAD MD 67, RTP NC 27711 Tel: 919-541-1585 fax -919-541-4017 shipping address RTF building ,2525 E.NC Hwy 54, Durham NC 27713 From: "Cammer, Michael" <[hidden email]> To: [hidden email] Date: 12/14/2011 03:44 PM Subject: odd inensity shift over time Sent by: Confocal Microscopy List <[hidden email]> ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear confocalers, We would appreciate help identifying (and remedying) a problem we have not encountered before. One of our users reported that the laser intensity of the confocal was dropping off during each of her imaging sessions. Today I put a mirror on the confocal and imaged for 4 1/2 hours in the reflectance mode with the slits to detect over than 20 nm open. During that time the intensities at 488, 543, and 633 nm rose to a peak between 15 and 60 minutes and then fell off at a slower rate (graph posted at http://www.flickr.com/photos/mcammer/6512342071/ ). Has anyone seen this pattern before and could help us identify the cause? Is it a laser issue (with three different lasers?), an AOTF issue, a detectors issue, a drifting out of alignment issue (although in this last case the resolution in fluorescent imaging continues to look fine)? Also, this is not a focus drift issue. The microscope is a Zeiss 710 with three spectral detectors. Any help identifying this greatly appreciated. ________________________________________________________ Michael Cammer, Assistant Research Scientist Skirball Institute of Biomolecular Medicine Lab: (212) 263-3208 Cell: (914) 309-3270 </PRE> <html> <body> ------------------------------------------------------------<br /> This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email.<br /> ================================= </body> </html> <PRE> |
|
|
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Thank you everybody for the suggestions. We ran a test overnight while recording room temperature. There were independent drifts in each wavelength's power and no correlation with room temperature. We're repeating the readings right now using the transmission detector. Based on your comments, I'm thinking the problem is the AOTF or its controller. Regardless, we filed a service request with Zeiss. I'll let you know the what happens. Regards, Michael ________________________________________________________ Michael Cammer, Assistant Research Scientist Skirball Institute of Biomolecular Medicine Lab: (212) 263-3208 Cell: (914) 309-3270 ------------------------------------------------------------ This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email. ================================= |
|
|
Ekaterina PAPUSHEVA-2 |
molecular beacons for RNA expression analysis
|
|
In reply to this post by Vladimir Ghukasyan-2
*****
To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi all, My question is not strictly confocal, but maybe you would have an idea.... we are considering using molecular beacons tp follow promoter activity in isolated cells from zebrafish embryos. We have problem cloning a GFP reporter for the gene expression (the recombinant promoter is not functional), therefore considering using molecular beacons for following mRNA concentration (mRNA sequence is known). I am a novice with molecular beacons, and have a beginners question: how does binding of a molecular beacon affect RNA lifecycle? Would not we fall in a trap, that the area of RNA that is bound to the beacon is not degraded=false positive signal persists? What are the controls for this? I would be very grateful if you could give a tip or recommend a literature! (while surfing myself) good weekend to everyone! Ekaterina ___ Ekaterina Papusheva, PhD Imaging Facility Manager Institute of Science and Technology Austria Phone +43-(0)2243 9000-1043; Mobile +43-(0)664 88509130 Fax +43-(0)2243 9000-2000 Am Campus 1 A-3400 Klosterneuburg Visit our website: www.ist.ac.at This message and any attachment (together "the Message") are confidential and intended only for the individual named. If you have received the Message in error you should not distribute or copy this Message. Please notify the sender immediately by e-mail if you have received this Message in error and delete the Message from your system. Correspondence via e-mail is for information purposes. IST Austria neither makes nor accepts legally binding statements unless explicitly otherwise agreed to the contrary. -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Vladimir Ghukasyan Sent: Mittwoch, 14. Dezember 2011 22:16 To: [hidden email] Subject: Re: odd inensity shift over time ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Michael, Does this happen when you run imaging with a single laser at a time? Vladimir On Wed, Dec 14, 2011 at 3:44 PM, Cammer, Michael <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear confocalers, > We would appreciate help identifying (and remedying) a problem we have not encountered before. > One of our users reported that the laser intensity of the confocal was dropping off during each of her imaging sessions. Today I put a mirror on the confocal and imaged for 4 1/2 hours in the reflectance mode with the slits to detect over than 20 nm open. During that time the intensities at 488, 543, and 633 nm rose to a peak between 15 and 60 minutes and then fell off at a slower rate (graph posted at http://www.flickr.com/photos/mcammer/6512342071/ ). > Has anyone seen this pattern before and could help us identify the cause? Is it a laser issue (with three different lasers?), an AOTF issue, a detectors issue, a drifting out of alignment issue (although in this last case the resolution in fluorescent imaging continues to look fine)? > Also, this is not a focus drift issue. > > The microscope is a Zeiss 710 with three spectral detectors. > > Any help identifying this greatly appreciated. > ________________________________________________________ > Michael Cammer, Assistant Research Scientist Skirball Institute of > Biomolecular Medicine > Lab: (212) 263-3208 Cell: (914) 309-3270 > > </PRE> > <html> > <body> > ------------------------------------------------------------<br /> > This email message, including any attachments, is for the sole use of > the intended recipient(s) and may contain information that is > proprietary, confidential, and exempt from disclosure under applicable > law. Any unauthorized review, use, disclosure, or distribution is > prohibited. If you have received this email in error please notify the > sender by return email and delete the original message. Please note, > the recipient should check this email and any attachments for the > presence of viruses. The organization accepts no liability for any > damage caused by any virus transmitted by this email.<br /> > ================================= </body> </html> <PRE> |
«
Return to Confocal Microscopy List
|
1 view|%1 views
| Free forum by Nabble | Edit this page |