Posted by
Zdenek Svindrych-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Trying-to-image-Cy7-5-AxioObserver-Z1-blocking-IR-tp7586976p7586986.html
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Michael,
there is a plastic filter holder BELOW the filter block. That is not in the
excitation light path. Our Axio setup is different, as we have an LSM 780
NLO coupled to the back port. But for a widefield setup it makes perfect
sense to include IR blocking filter somewhere in the lamp or in the
illumination path, for the fluorescence filters are often misbehaving in the
IR...
You can grab a (metric) screwdriver and explore for yourself, or (as
suggested several times) ask Zeiss.
Good luck!
zdenek
---------- Původní e-mail ----------
Od: Cammer, Michael <
[hidden email]>
Komu:
[hidden email]
Datum: 12. 7. 2017 9:30:29
Předmět: Re: Trying to image Cy7.5 -- AxioObserver.Z1 blocking IR?
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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
________________________________________
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=j5oPpO0eBH1
iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&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=j5oPpO0eBH1iio48DtsedbOBGmuw5
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FA&e=
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