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cy7 nuclear stain for live cell microscopy

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Zervantonakis, Ioannis Zervantonakis, Ioannis
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cy7 nuclear stain for live cell microscopy

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Any recommendations for Cy5.5 or Cy7 nuclear stains? We have tried amine-reactive dyes but these cause autofluorescence, so we are looking for dyes similar to TOPRO-5, but this is no longer available from Thermo.




______________________

Ioannis K. Zervantonakis

Assistant Professor

Dept. of Bioengineering

U. Pittsburgh Swanson School of Engineering

UPMC Hillman Cancer Center

(e) [hidden email]<mailto:[hidden email]>

(w) www.zervalab.com<http://www.zervalab.com/>

(p) 4126484442
Zdenek Svindrych-2 Zdenek Svindrych-2
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Re: cy7 nuclear stain for live cell microscopy

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Hi Ioannis, have you thought about DRAQ5, DRAQ7?
Best, zdenek

On Mon, Nov 9, 2020 at 5:05 PM Zervantonakis, Ioannis <[hidden email]> 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.
> *****
>
> Any recommendations for Cy5.5 or Cy7 nuclear stains? We have tried
> amine-reactive dyes but these cause autofluorescence, so we are looking for
> dyes similar to TOPRO-5, but this is no longer available from Thermo.
>
>
>
>
> ______________________
>
> Ioannis K. Zervantonakis
>
> Assistant Professor
>
> Dept. of Bioengineering
>
> U. Pittsburgh Swanson School of Engineering
>
> UPMC Hillman Cancer Center
>
> (e) [hidden email]<mailto:[hidden email]>
>
> (w) www.zervalab.com<http://www.zervalab.com/>
>
> (p) 4126484442
>


--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth
George McNamara George McNamara
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Re: cy7 nuclear stain for live cell microscopy

In reply to this post by Zervantonakis, Ioannis
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DNA: Zombie NIR from BioLegend, excites ok at ~640nm according to
https://searchlight.semrock.com

RNA (since nucle have RNA): various SYTO etc dyes from ThermoFisher.

Your new favorite nuclear protein: could use an antibody with some NIR
fluorophore (Alexa Fluor 750, Brilliant Violet 750) to detect any of:
histone, Lamin (A/C or B), nuclear pore complex (NPC), etc. NPC provide
nice punctate spots (microscopy) or standard test targets (nanoscopy),
avoids massive blob of DNA, RNA or histone, so simpler for compartment
segmentation. Going for NPC could detect using the anti-TPR and
fluorescent 2ndary nanobody used in Pleiner 2018 JCB
(https://doi.org/10.1083/jcb.201709115 ... 2ndary nanobodies now
available from nano-tag.com. chromotek.com or ThermoFisher).


On 11/9/2020 4:55 PM, Zervantonakis, Ioannis 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.
> *****
>
> Any recommendations for Cy5.5 or Cy7 nuclear stains? We have tried amine-reactive dyes but these cause autofluorescence, so we are looking for dyes similar to TOPRO-5, but this is no longer available from Thermo.
>
>
>
>
> ______________________
>
> Ioannis K. Zervantonakis
>
> Assistant Professor
>
> Dept. of Bioengineering
>
> U. Pittsburgh Swanson School of Engineering
>
> UPMC Hillman Cancer Center
>
> (e) [hidden email]<mailto:[hidden email]>
>
> (w) www.zervalab.com<http://www.zervalab.com/>
>
> (p) 4126484442
Cammer, Michael-2 Cammer, Michael-2
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Re: cy7 nuclear stain for live cell microscopy

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Post images on http://www.imgur.com and include the link in your posting.
*****

A warning that we've had focus problems with IR probes for non-multiphoton with lenses that are not chromatically corrected out this far into red.

Cheers-

Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory

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

[hidden email]<mailto:[hidden email]>  http://nyulmc.org/micros  http://microscopynotes.com/

Voice direct only, no text or messages:  1-914-309-3270 and 1-646-501-0567



________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of George McNamara <[hidden email]>
Sent: Monday, November 9, 2020 5:47 PM
To: [hidden email]
Subject: Re: cy7 nuclear stain for live cell microscopy

[EXTERNAL]

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Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwICaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=W0rfKZVtl7OPRmYLCXKPtqQqXn66gzmke8zCWN36C70&s=sYD_NY4DDccfvFzYuHAuN5MMiCm9OxMvOcelaUEc6hI&e=  and include the link in your posting.
*****

DNA: Zombie NIR from BioLegend, excites ok at ~640nm according to
https://urldefense.proofpoint.com/v2/url?u=https-3A__searchlight.semrock.com&d=DwICaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=W0rfKZVtl7OPRmYLCXKPtqQqXn66gzmke8zCWN36C70&s=nOF-3Zo5toBXGFCEhkBDgVC3-iXTAUt9JMJaBVVdumI&e=

RNA (since nucle have RNA): various SYTO etc dyes from ThermoFisher.

Your new favorite nuclear protein: could use an antibody with some NIR
fluorophore (Alexa Fluor 750, Brilliant Violet 750) to detect any of:
histone, Lamin (A/C or B), nuclear pore complex (NPC), etc. NPC provide
nice punctate spots (microscopy) or standard test targets (nanoscopy),
avoids massive blob of DNA, RNA or histone, so simpler for compartment
segmentation. Going for NPC could detect using the anti-TPR and
fluorescent 2ndary nanobody used in Pleiner 2018 JCB
(https://urldefense.proofpoint.com/v2/url?u=https-3A__doi.org_10.1083_jcb.201709115&d=DwICaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=W0rfKZVtl7OPRmYLCXKPtqQqXn66gzmke8zCWN36C70&s=I8UNsE1Q3-PGV4Xk74nmGj5F4oOXGaU-bqFGQBwMqns&e=  ... 2ndary nanobodies now
available from nano-tag.com. chromotek.com or ThermoFisher).


On 11/9/2020 4:55 PM, Zervantonakis, Ioannis wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwICaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=W0rfKZVtl7OPRmYLCXKPtqQqXn66gzmke8zCWN36C70&s=W0EIoz4CvelYUWxXaRYLUuZidQXr9vEWshl6mPB4Yj4&e=
> Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwICaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=W0rfKZVtl7OPRmYLCXKPtqQqXn66gzmke8zCWN36C70&s=sYD_NY4DDccfvFzYuHAuN5MMiCm9OxMvOcelaUEc6hI&e=  and include the link in your posting.
> *****
>
> Any recommendations for Cy5.5 or Cy7 nuclear stains? We have tried amine-reactive dyes but these cause autofluorescence, so we are looking for dyes similar to TOPRO-5, but this is no longer available from Thermo.
>
>
>
>
> ______________________
>
> Ioannis K. Zervantonakis
>
> Assistant Professor
>
> Dept. of Bioengineering
>
> U. Pittsburgh Swanson School of Engineering
>
> UPMC Hillman Cancer Center
>
> (e) [hidden email]<mailto:[hidden email]>
>
> (w) https://urldefense.proofpoint.com/v2/url?u=http-3A__www.zervalab.com&d=DwICaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=W0rfKZVtl7OPRmYLCXKPtqQqXn66gzmke8zCWN36C70&s=RkssCIKMDXIkx7q4BWM6qR_QTF9UJiBp6NbNI946RLY&e= <https://urldefense.proofpoint.com/v2/url?u=http-3A__www.zervalab.com_&d=DwICaQ&c=j5oPpO0eBH1iio48DtsedeElZfc04rx3ExJHeIIZuCs&r=hUBj2D5n6oKThx2L01qn8IORZb5f-ruLVXPmQ1zQNnM&m=W0rfKZVtl7OPRmYLCXKPtqQqXn66gzmke8zCWN36C70&s=LKygFkUZCYIrtkkh85BdD4Gj9V6va5i-CLbEAcq1xjQ&e= >
>
> (p) 4126484442

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Angela Fontaine Angela Fontaine
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Re: cy7 nuclear stain for live cell microscopy

In reply to this post by Zervantonakis, Ioannis
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Hi Ioannis,

have you tried the SYTO and SYTOX dyes (also form Thermo).
I was very happy with the result - I used them to label in vivo and did
intravital imaging with a multiphoton microscope to visualize Netosis in
vivo.

Best wishes,
Angela

On Tue, 10 Nov 2020 at 09:05, Zervantonakis, Ioannis <[hidden email]> 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.
> *****
>
> Any recommendations for Cy5.5 or Cy7 nuclear stains? We have tried
> amine-reactive dyes but these cause autofluorescence, so we are looking for
> dyes similar to TOPRO-5, but this is no longer available from Thermo.
>
>
>
>
> ______________________
>
> Ioannis K. Zervantonakis
>
> Assistant Professor
>
> Dept. of Bioengineering
>
> U. Pittsburgh Swanson School of Engineering
>
> UPMC Hillman Cancer Center
>
> (e) [hidden email]<mailto:[hidden email]>
>
> (w) www.zervalab.com<http://www.zervalab.com/>
>
> (p) 4126484442
>

--
 <https://www.centenary.org.au>
Steffen Dietzel Steffen Dietzel
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Re: cy7 nuclear stain for live cell microscopy

In reply to this post by Zervantonakis, Ioannis
*****
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Post images on http://www.imgur.com and include the link in your posting.
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Ioannis,

sounds like SiR-DNA or SiR700-DNA might be working for you. Worked nice
in our hands. Spirochrome also has some new SPY dyes that might do the
trick, but we have no experience yet with those.

https://spirochrome.com/

Best

Steffen

Am 09.11.2020 um 22:55 schrieb Zervantonakis, Ioannis:

> *****
> 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.
> *****
>
> Any recommendations for Cy5.5 or Cy7 nuclear stains? We have tried amine-reactive dyes but these cause autofluorescence, so we are looking for dyes similar to TOPRO-5, but this is no longer available from Thermo.
>
>
>
>
> ______________________
>
> Ioannis K. Zervantonakis
>
> Assistant Professor
>
> Dept. of Bioengineering
>
> U. Pittsburgh Swanson School of Engineering
>
> UPMC Hillman Cancer Center
>
> (e) [hidden email]<mailto:[hidden email]>
>
> (w) www.zervalab.com<http://www.zervalab.com/>
>
> (p) 4126484442

--
------------------------------------------------------------
Steffen Dietzel, PD Dr. rer. nat
Ludwig-Maximilians-Universität München
Biomedical Center (BMC)
Head of the Core Facility Bioimaging

Großhaderner Straße 9
D-82152 Planegg-Martinsried
Germany

http://www.bioimaging.bmc.med.uni-muenchen.de
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