Live cell imaging in mouse embryonic stem cells

[Alivia]

What are the dyes and medium that I should use if I were to perform a live-cell imaging on mouse embryonic stem cell to track the cell division?

[c.surridge]

Alivia,

I am sure that others on this group will have first hand experience to share with you. There are also a number of Protocols published by us that may be helpful. This one looks particularly appropriate for your problem:

Embryonic and induced pluripotent stem cell staining and sorting with the live-cell fluorescence imaging probe CDy1

• Nam-Young Kang,
 
• Seong-Wook Yun,
 
• Hyung-Ho Ha,
 
• Sung-Jin Park &
 
• Young-Tae Chang

Nature Protocols 6, 1044-1052 doi:10.1038/nprot.2011.350

Also worth looking at may be

Generation of eggs from mouse embryonic stem cells and induced pluripotentstem cells

• Katsuhiko Hayashi &
 
• Mitinori Saitou

Nature Protocols 8, 1513-1524 doi:10.1038/nprot.2013.090

Alternatively another approach you could try is using reporters as is described here:

Long term non-invasive imaging of embryonic stem cells using reporter genes

• Ning Sun,
 
• Andrew Lee &
 
• Joseph C Wu

Nature Protocols 4, 1192-1201 doi:10.1038/nprot.2009.100

Good luck with your experiments.

Chris

(Chief Editor, Nature Protocols)

[Alivia]

Thank you Chris for you reply! 

Unfortunately I only have a few months left to optimize and work this out so I am actually looking at simpler methods, eg. Hoechst stain.

I understand that Hoechst stain may affect cell division due to its intrinsic activity.  

Greatly appreciate if any researchers have first hand experience handling similar work.

[c.surridge]

I hope we can find you an answer. 

I don't know that this is quite what you want but this Protocol uses Hoechst 33342 stain on hPSCs.

Adapting human pluripotent stem cells to high-throughput and high-content screening

Sabrina C Desbordes and Lorenz Studer

Nature Protocols

 

8

 

p111 - p130

 

20/12/2012

doi:10.1038/nprot.2012.139

Chris