http://confocal-microscopy-list.275.s1.nabble.com/Trying-to-image-Cy7-5-AxioObserver-Z1-blocking-IR-tp7586976p7586987.html
aperture diaphragm in the rear port. This would be internal to the body of
have an even worse 50% cut-off at 716 nm. Light Googling can't seem to
good news if you're trying to illuminate with > 720 nm. In any case, a
> *****
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>
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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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> umn.edu_cgi-2Dbin_
> wa-3FA0-3Dconfocalmicroscopy&d=DQIFaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5
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> hwRGJZNcWAhP
> 78SlBGjpWK7aUX3i2m17ECXlq3M4bCw&s=DASw7UwNEEnOcBauPfKePh4akaFo79
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> Post images on
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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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> Post images on
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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 &
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> hwRGJZNcWAhP
> 78SlBGjpWK7aUX3i2m17ECXlq3M4bCw&s=YfHG4GsP2w6AWO_eV_YxwEJd0_
> -WuRVVzQoRmEHD-
> FA&e=
>
>
>
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