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mcammer on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Trying-to-image-Cy7-5-AxioObserver-Z1-blocking-IR-tp7586976p7586985.html
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Thank you for the replies. Thank you for the diagram which I reposted at
http://microscopynotes.com/axioobserver/IR/diagram.png with the part of the light path we're having a problem with.
The light guide is plugged in to the fluorescence illumination optics at the back of the AxioObserver. Then there is a 50/50 mirror in the filter block position to project to the back aperture of the lens. We put the mirror in because the electronics of the AxioObserver won't allow the shutter to open if the side port is open which we would need to put the power meter probe in. But we tested the mirror independently and it works ok out to 770 nm at least, so the problem must be in the illumination optics path. But Sven points out there may be a filter between the filter block and the objective, so I will look for that, but since we have the IR autofocus device, I don't see how this would be possible.
I'll try another scope, but this is the one we really need to use because of the camera and stage for tiling, neither which can be moved to another scope body. Also, we cannot do customization to this stand that would prevent routine use because it is in microscopy core. Hence why we're looking for a filter that might be simple to pop out.
Thanks.
=*===========================================================*=
Michael Cammer, DART Microscopy Laboratory, NYU Langone Medical Center
Cell: 914-309-3270 (this is for calling, not texting) Office: Skirball 2nd Floor main office, back right
http://ocs.med.nyu.edu/microscopy &
http://microscopynotes.com/-----Original Message-----
From: Cammer, Michael
Sent: Tuesday, July 11, 2017 10:31 PM
To: Confocal Microscopy List
Subject: RE: Trying to image Cy7.5 -- AxioObserver.Z1 blocking IR?
At this point, there are no filters (that we know of) in the light path. The light guide is plugged in to the fluorescence illumination optics at the back of the AxioObserver. Then there is a 50/50 mirror in the filter block position to project to the back aperture of the lens. We put the mirror in because the electronics of the AxioObserver won't allow the shutter to open if the side port is open which we would need to put the power meter probe in. But the mirror is working ok out to 770 nm at least, so I thought issue must be in the illumination optics path. But Sven points out there may be a filter between the filter block and the objective, so I will look for that, but since we have the IR autofocus device, I don;t see how this would be possible.
Thanks.
_________________________________________
Michael Cammer, Optical Microscopy Specialist
http://ocs.med.nyu.edu/microscopy http://microscopynotes.com/Cell: (914) 309-3270
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From: Confocal Microscopy List [
[hidden email]] on behalf of Sven Terclavers [
[hidden email]]
Sent: Tuesday, July 11, 2017 10:25 PM
To:
[hidden email]
Subject: Re: Trying to image Cy7.5 -- AxioObserver.Z1 blocking IR?
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The light train itself has a transmission beyond 1100nm. I assume the (optional) IR filter you're talking about is the one that sits just below the filter cubes in a slider, right above the tube lens. This one should not even affect illumination as it's a filter for emission light. Have you checked whether there's a small compensation glass sitting on top of the filter turret? I don't know its specific transmission spectrum, but it might be that one. Since you have DF you can leave this out completely. Furthermore, no attenuator filters that could influence transmission? And as mentioned by Craig, which filters are you using? When you remove the filter turret, do you see the light coming through the light train?
Best regards,
Sven
Sent from Mail for Windows 10
From: Cammer, Michael
Sent: Tuesday, July 11, 2017 10:01 PM
To:
[hidden email]
Subject: Trying to image Cy7.5 -- AxioObserver.Z1 blocking IR?
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A few weeks ago we asked about finding a light source at 780ish nm for exciting Cy7.5. As always, people on this listserv provided very helpful suggestions. We were able to get a pE-4000 system with multiple LEDs but are finding that the Zeiss AxioObserver.Z1 fluorescence illumination optics appear to be cutting wavelengths above 700 nm.
We adjusted the LED powers such that 635, 660, 740, and 770 nm all have 52 mW at the end of the liquid light guide. We removed the IR filter for the autofocus system that is in the objectives nosepiece. However, the light at 740 and 770 nm is blocked from arriving at the objective lenses.
Does anyone know whether there is a filter somewhere else in the light path we could easily remove to pass these wavelengths or is this a limitation of the light train?
(graphs of the power loss posted at
https://urldefense.proofpoint.com/v2/url?u=http-3A__microscopynotes.com_axioobserver_IR_&d=DQIFaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=hwRGJZNcWAhP78SlBGjpWK7aUX3i2m17ECXlq3M4bCw&s=5H18aiZASsoKyCJ0CWz_vELNL-F_nJdbJz3htY0tpT8&e= ) Thank you!!
=*===========================================================*=
Michael Cammer, DART Microscopy Laboratory, NYU Langone Medical Center
Cell: 914-309-3270 (this is for calling, not texting) Office: Skirball 2nd Floor main office, back right
http://ocs.med.nyu.edu/microscopy &
https://urldefense.proofpoint.com/v2/url?u=http-3A__microscopynotes.com_&d=DQIFaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=hwRGJZNcWAhP78SlBGjpWK7aUX3i2m17ECXlq3M4bCw&s=YfHG4GsP2w6AWO_eV_YxwEJd0_-WuRVVzQoRmEHD-FA&e=------------------------------------------------------------
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