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I am waiting to hear from Zeiss to find out whether we can easily remove the heat filter(s) in the new Axioobserver.
In the meantime, I found a Zeiss AxioPlan, removed the KG? filter from the back, and the 740 and 770 nm light goes through it fine.
Thank you all for the feedback.
=*===========================================================*=
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: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Rusty Nicovich
Sent: Wednesday, July 12, 2017 5:51 PM
To: [log in to unmask]
Subject: Re: Trying to image Cy7.5 -- AxioObserver.Z1 blocking IR?
From my schematic, the Axio Observer has a heat filter right behind the aperture diaphragm in the rear port. This would be internal to the body of the scope and unlikely to be accessible or removable.
Assuming it's this part number as listed on the Zeiss website:
https://urldefense.proofpoint.com/v2/url?u=https-3A__www.micro-2Dshop.zeiss.com_index.php-3Fp-3Dat-26f-3Da-26i-3D10051-5F400000001200100000-26o-26h-3D25-26n-3D0-26sd-3D467830-2D0000-2D000-26l-3Den&d=DQIFaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=2dd3bRi4R-tIWosCp6IoMx8t8cNYhoiAng6VNFWuWi0&s=v-XN3q2ZaJvuNm5ZaAEqtVQ_q4XN3I3FdCy07-lt2HQ&e=
and it's really typical KG 1 glass, this page has the transmission/blocking
curves:
https://urldefense.proofpoint.com/v2/url?u=https-3A__www.ahf.de_art-2DWaermeschutzfilter-5FKG-5F1-5F-5FD-5F25-5Fmm-3BF01-2D125.html&d=DQIFaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=2dd3bRi4R-tIWosCp6IoMx8t8cNYhoiAng6VNFWuWi0&s=-qcTfxL57QX1KanZkMOFlMP6K4tsQrLDoibE5l55Fb4&e=
The 50% cut-off for transmission is 740 nm, which may be bad news if you are illuminating with 740 nm or 770 nm.
These KG 1 optics:
https://urldefense.proofpoint.com/v2/url?u=https-3A__www.knightoptical.com_stock_optical-2Dcomponents_uvvisnir-2Doptics_filters_short-2Dpass-2Dfilters_kg1-2Dheat-2Dglass-2Dfilters_&d=DQIFaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=2dd3bRi4R-tIWosCp6IoMx8t8cNYhoiAng6VNFWuWi0&s=8OjWDiwz0JAjfQYGPKj0iraF1qY_P0R96tCz8XuBUls&e=
have an even worse 50% cut-off at 716 nm. Light Googling can't seem to find more info on the Zeiss product transmission but it doesn't look like good news if you're trying to illuminate with > 720 nm. In any case, a call to Zeiss is going to be the way to go here.
Thanks,
Rusty
On Wed, Jul 12, 2017 at 7:04 AM, <[log in to unmask]> wrote:
> *****
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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 <[log in to unmask]>
> Komu: [log in to unmask]
> 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
> https://urldefense.proofpoint.com/v2/url?u=http-3A__microscopynotes.co
> m_axioobserver_IR_diagram.png&d=DQIFaQ&c=j5oPpO0eBH1iio48DtsedbOBGmuw5jHLjgvtN2r4ehE&r=oU_05LztNstAydlbm5L5GDu_vAdjXk3frDLx_CqKkuo&m=2dd3bRi4R-tIWosCp6IoMx8t8cNYhoiAng6VNFWuWi0&s=dqDcUpwkpA9gx4zh4bYfwfALTKNv8TvTKy_XRIapAWk&e= 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 &
> https://urldefense.proofpoint.com/v2/url?u=http-3A__microscopynotes.co
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> 6VNFWuWi0&s=QZ4D6s7slhBbAFuZqIi_9Wc5cwUtkaI-MnjamzzrURY&e=
>
>
>
> -----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
> https://urldefense.proofpoint.com/v2/url?u=http-3A__microscopynotes.co
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> Cell: (914) 309-3270
>
> ________________________________________
> From: Confocal Microscopy List [[log in to unmask]] on
> behalf of Sven Terclavers [[log in to unmask]]
> Sent: Tuesday, July 11, 2017 10:25 PM
> To: [log in to unmask]
> 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: [log in to unmask]
> 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=j5oPpO0
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> 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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>
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