Adipose tissue in flow cytometry

Lora Barsky lbarsky at chla.usc.edu
Fri Jun 8 15:34:22 EDT 2007


ONce you have your adipose seperated, you can positively select for  
it using Nile Red dye (Invitrogen).

Nile Red Dye
Hydrophobic probe
9-diethylamino-5H-benzo[alpha]phenoxazine-5-one
EM MAX is determined by the relative hydrophobicity of surrounding  
environment
Dissolved in neutral lipids, emits gold (FL2)
Dissolved in amphipathic lipids, emits red (FL3)
Smyth and Wharton (1992) characterized the differentiation of adipocytes
Ratio of Gold/Red defines differentiation

See papers:
Berstein, RL etal. Cytometery 10:469-474 (1989)
Backesjo, C-M, etal. Journal of Bone and Miner Research  Vol 21,  
Number 7, 2006 page 993-1002
Kelley and Gimble, Endocrinology vol 139, no 5 (1998)
Greenspan 1985 Journal of Cell Biology Vol 100: Marach	1985  965-973
Smith and Wharton, Exp Cell Research 199, 29-38 (1992)

good luck,
Lora


Lora Barsky
Manager, Flow Cytometry Lab
Division of Research Immunology/BMT/GISCT Program
Childrens Hospital Los Angeles
4650 Sunset Blvd., MailStop #62
Los Angeles, CA  90027

lbarsky at chla.usc.edu
323-361-5935
323-361-8185 FAX


On Jun 5, 2007, at 10:26 AM, Ann Kelly wrote:

> Hello Flow-ers,
>    My principal investigator is looking for information from those  
> researchers with experience in the analysis of adipose tissue via  
> flow cytometry. This is a new area of research for our group, and  
> we are seeking a method for cell separation. Our hope is to be able  
> to isolate lymphocytes and granulocytes from the whole of the  
> adipose tissue.  Here are a couple of basic questions:
>  1. What method of tissue digestion is favored prior to a cell sort?
>  2. Do we need to sort the granulocytes and lymphocytes from the  
> other cells present in adipose tissue with a density gradient  
> ( i.e. sucrose, Percoll, etc.) prior to performing a cell sort?
>
> Any advice about our new endeavor is welcome.
>
> Thank you,
> Ann Kelly
> Oregon Health Science University
> Senior Research Assistant
> Department of Pulmonary and Critical Care Medicine
>

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