Detection of near infrared

[Marcel Mettlen]

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Hello - I was wondering which make/model detectors (InP/InGaAs PMT, sCMOS camera, ....) are currently best suited to detect the near infrared emission from Cy7, Cy7.8, IRDye800, etc?
Thanks,
Marcel Mettlen
-------------------------
Res. Assistant Professor and
Director of Research and Collaborations
UT Southwestern Medical Center
Dept of Cell Biology
Dallas, TX, USA

[Michael Giacomelli-2]

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Hi Marcel,

For operation at 650-800nm, silicon detectors are generally the most sensitive.  For 2D sensors, an sCMOS camera (~70% QE, few photon sensitivity) is probably the best you can do.  If you need point detectors like for confocal, silicon photomultipliers (15-30% QE, although they drop off rapidly in this range) are the most sensitive option, followed by GaAs PMTs (5-15% QE depending on how you define it).

As for which is better in an absolute sense, sCMOS sensors are usually not shot noise limited for the lowest light flux detected, but they have much higher QE and so detect a lot more photons.  It will probably depend on the exact wavelengths and light fluxes you are looking at, although I haven't done the math.

Mike

On Wed, Jan 29, 2020 at 8:24 AM Marcel Mettlen <[hidden email]<mailto:[hidden email]>> wrote:
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Hello - I was wondering which make/model detectors (InP/InGaAs PMT, sCMOS camera, ....) are currently best suited to detect the near infrared emission from Cy7, Cy7.8, IRDye800, etc?
Thanks,
Marcel Mettlen
-------------------------
Res. Assistant Professor and
Director of Research and Collaborations
UT Southwestern Medical Center
Dept of Cell Biology
Dallas, TX, USA


--
Michael Giacomelli, Ph.D.
Assistant Professor
Department of Biomedical Engineering
University of Rochester

[Andrea Latini]

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** no commercial interest **

Dear Marcel,
in addition to Mike's comments let me remind you that in case you'd select a CCD system you must choose a Deep Depletion model to avoid etaloning artifact effect.
Deep Depleted CCDs exhibit a >80% QE between 600nm - 900nm
ref:
Andor https://andor.oxinst.com/learning/view/article/optical-etaloning-in-charge-coupled-devices 
Horiba: https://www.horiba.com/fileadmin/uploads/Scientific/Documents/OSD/OSD-Etalon_Fringe_Suppression.pdf
Princeton - Teledyne: https://www.princetoninstruments.com/userfiles/files/technotes/eXcelon-technote-Novel-performance-enhancing-CCD-technology.pdf

Andrea Latini, CEO
CrestOptics SpA

[George McNamara]

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On point scanning confocal >712nm GaAs PMT -- or GaAs faceplate hybrid
detector -- is likely the way to go, see ~3/4 down page at

http://confocal.jhu.edu/current-equipment/fv3000/

for QE curves and info on our FV3000RS (two GaAs PMTs, current
beamsplitter and emission filters below graph). Hamamatsu electronics
identical (nominally anyway) for the GaAsP internal (HSD) detectors and
GaAs on our FV3000RS.

APDs have outstanding QE (~80%) but typically low photon counting rate
(confocal microscopy is often above the max count rate, or would take
too many line accumulation to be useful).

***

camera ... this is 2020, so back illuminated sCMOS likely more useful
than (typically bigger, fewer pixels, slow readout) b.i. EMCCD, if that
is what you need. More cooled, better (usually ... cost being main limit).

happy, 2020,

George


On 1/29/2020 8:13 AM, Marcel Mettlen wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Hello - I was wondering which make/model detectors (InP/InGaAs PMT, sCMOS camera, ....) are currently best suited to detect the near infrared emission from Cy7, Cy7.8, IRDye800, etc?
> Thanks,
> Marcel Mettlen
> -------------------------
> Res. Assistant Professor and
> Director of Research and Collaborations
> UT Southwestern Medical Center
> Dept of Cell Biology
> Dallas, TX, USA

[Zbigniew Mikulski]

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Hi Marcel!

We added a Cy7-like filter cube to our AxioScan Z1 which runs on OrcaFlash 4.0 v2 and Colibri7 LED excitation. This is a great combination for imaging of tissues as you get very little autofluorescence in this region. There is not a ton of reagents available in this range, we are using secondary antibodies conjugated to CF750 from Biotium.

Some juicy tonsil image for you
https://imgur.com/r9Zyl6S

Best, Z.

Zbigniew Mikulski, PhD
Director, Microscopy Core Facility, Instructor | La Jolla Institute for Immunology | 9420 Athena Circle, La Jolla, CA 92037

[Cammer, Michael]

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If using a CCD camera with longer IR probes and you have autofocus, you may need to remove the autofocus optics to pass 790ish nm light to camera.  We were doing a custom project with a chemistry group with probes that excited around 780 nm and emitted 800 nm+ and it worked widefield with a CCD camera after we removed the autofocus optics.


Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory

NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY  10016

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________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Zbigniew Mikulski <[hidden email]>
Sent: Thursday, January 30, 2020 9:22:31 PM
To: [hidden email]
Subject: Re: Detection of near infrared

[EXTERNAL]

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Hi Marcel!

We added a Cy7-like filter cube to our AxioScan Z1 which runs on OrcaFlash 4.0 v2 and Colibri7 LED excitation. This is a great combination for imaging of tissues as you get very little autofluorescence in this region. There is not a ton of reagents available in this range, we are using secondary antibodies conjugated to CF750 from Biotium.

Some juicy tonsil image for you
https://urldefense.proofpoint.com/v2/url?u=https-3A__imgur.com_r9Zyl6S&d=DwIFaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=Ks-Zps7B-35Ewt5RsFvOH4lRYI21BBMmrgZzKsR2l3Y&s=Z2EyqvUuUdhvEofy0h751YpnqBndMJf1XyY999isYv4&e=

Best, Z.

Zbigniew Mikulski, PhD
Director, Microscopy Core Facility, Instructor | La Jolla Institute for Immunology | 9420 Athena Circle, La Jolla, CA 92037