clearing methods and Abberior dyes

[Ekaterina PAPUSHEVA-2]

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Dear all,

Does someone have experience with clearing samples (brain tissue, lymph nodes) stained with far-red abberior dyes?
What works best? How does IDisco perform?

Thanks in advance,
Ekaterina


___

Ekaterina Papusheva

Bioimaging Facility

Institute of Science and Technology Austria

Phone  [phone];
Mobile [phone]
Fax    [phone]

Am Campus 1

A-3400 Klosterneuburg

Visit our website: www.ist.ac.at<http://www.ist.ac.at/>


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[Douglas Richardson]

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Hello Ekaterina,

With the exception of high concentrations of TDE (which results in rapid
bleaching of blue/green dyes but enhances fluorescence from red cyanine
dyes - Staudt et al.MRT 2007), I have not found any clearing solutions that
strongly suppress fluoresce from organic dye molecules.  However, all
clearing methods (and fixation methods) have to potential to alter antibody
binding sites and reduce/eliminate the ability of your primary antibody to
bind its target.

Additionally, your cleared samples are often not in an oxygen scavenging
environment (unlike when using an antifade mounting media), therefore you
may observe more rapid bleaching. Most Atto dyes are quite stable, and if
you're using lightsheet microscopy to image the sample, this likely won't
be a problem.

-Doug


On Wed, Oct 24, 2018 at 4:06 AM Ekaterina PAPUSHEVA <
[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.
> *****
>
> Dear all,
>
> Does someone have experience with clearing samples (brain tissue, lymph
> nodes) stained with far-red abberior dyes?
> What works best? How does IDisco perform?
>
> Thanks in advance,
> Ekaterina
>
>
> ___
>
> Ekaterina Papusheva
>
> Bioimaging Facility
>
> Institute of Science and Technology Austria
>
> Phone  [phone];
> Mobile [phone]
> Fax    [phone]
>
> Am Campus 1
>
> A-3400 Klosterneuburg
>
> Visit our website: www.ist.ac.at<http://www.ist.ac.at/>
>
>
> This message and any attachment (together "the Message") are confidential
> and intended only for the individual named. If you have received the
> Message in error you should not distribute or copy this Message. Please
> notify the sender immediately by e-mail if you have received this Message
> in error and delete the Message from your system. Correspondence via e-mail
> is for information purposes. IST Austria neither makes nor accepts legally
> binding statements unless explicitly otherwise agreed to the contrary.
>

[Ekaterina PAPUSHEVA-2]

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Thank you very much, Doug!

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Douglas Richardson
Sent: Wednesday, October 24, 2018 11:46 PM
To: [hidden email]
Subject: Re: clearing methods and Abberior dyes

*****
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*****

Hello Ekaterina,

With the exception of high concentrations of TDE (which results in rapid
bleaching of blue/green dyes but enhances fluorescence from red cyanine
dyes - Staudt et al.MRT 2007), I have not found any clearing solutions that
strongly suppress fluoresce from organic dye molecules.  However, all
clearing methods (and fixation methods) have to potential to alter antibody
binding sites and reduce/eliminate the ability of your primary antibody to
bind its target.

Additionally, your cleared samples are often not in an oxygen scavenging
environment (unlike when using an antifade mounting media), therefore you
may observe more rapid bleaching. Most Atto dyes are quite stable, and if
you're using lightsheet microscopy to image the sample, this likely won't
be a problem.

-Doug


On Wed, Oct 24, 2018 at 4:06 AM Ekaterina PAPUSHEVA <
[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.
> *****
>
> Dear all,
>
> Does someone have experience with clearing samples (brain tissue, lymph
> nodes) stained with far-red abberior dyes?
> What works best? How does IDisco perform?
>
> Thanks in advance,
> Ekaterina
>
>
> ___
>
> Ekaterina Papusheva
>
> Bioimaging Facility
>
> Institute of Science and Technology Austria
>
> Phone  [phone];
> Mobile [phone]
> Fax    [phone]
>
> Am Campus 1
>
> A-3400 Klosterneuburg
>
> Visit our website: www.ist.ac.at<http://www.ist.ac.at/>
>
>
> This message and any attachment (together "the Message") are confidential
> and intended only for the individual named. If you have received the
> Message in error you should not distribute or copy this Message. Please
> notify the sender immediately by e-mail if you have received this Message
> in error and delete the Message from your system. Correspondence via e-mail
> is for information purposes. IST Austria neither makes nor accepts legally
> binding statements unless explicitly otherwise agreed to the contrary.
>