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Sven Terclavers-3 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Trying-to-image-Cy7-5-AxioObserver-Z1-blocking-IR-tp7586976p7586978.html
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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
http://microscopynotes.com/axioobserver/IR/ )
Thank you!!
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Michael Cammer, DART Microscopy Laboratory, NYU Langone Medical Center
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