Zebrafish apoptosis in vivo - staining for time-lapse imaging

[Leonor Morgado]

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Dear all,

We are aiming to do time-lapse imaging of apoptotic cells in zebrafish. We
are browsing stainings that are specific for apoptotic cells and we would
appreciate any recommendation.

We will start with acridine orange - is there any precaution we should take
account for?
Did anyone worked with SYTOX deep red or TO-PRO3 for this purpose and got
good results?
Any other staining that you would recommend? Ideally with an emission in
green or far red range.

Thank you very much in advance,

Leonor Morgado
Champalimaud ABBE Platform
Advanced BioImaging and BioOptics Experimental Platform
Research Technician

[Gaurav Joshi]

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Hi Leonor,

I have used cell event green caspase -3/7 from ThermoFisher for live cell imaging with great success. Activation of caspase -3 cleaves the substrate -DEVD leading to fluorescence of the probe. The fluorescence is prominent in the nucleus making it easy to quantify the cells.

https://www.thermofisher.com/order/catalog/product/C10423#/C10423

FYI - No commercial interest. There are other companies selling a similar reagent.

Best,
Gaurav.

Gaurav Joshi, PhD  |  Imaging Specialist
Integrated Cellular Imaging (ICI), Emory University
Health Sciences Research Building (HSRB)
EG72, 1760 Haygood Drive, Atlanta, GA 30322
ici.emory.edu

         
 

On 12/2/19, 1:05 PM, "Confocal Microscopy List on behalf of Leonor Morgado" <[hidden email] on behalf of [hidden email]> wrote:

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    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,
   
    We are aiming to do time-lapse imaging of apoptotic cells in zebrafish. We
    are browsing stainings that are specific for apoptotic cells and we would
    appreciate any recommendation.
   
    We will start with acridine orange - is there any precaution we should take
    account for?
    Did anyone worked with SYTOX deep red or TO-PRO3 for this purpose and got
    good results?
    Any other staining that you would recommend? Ideally with an emission in
    green or far red range.
   
    Thank you very much in advance,
   
    Leonor Morgado
    Champalimaud ABBE Platform
    Advanced BioImaging and BioOptics Experimental Platform
    Research Technician