RE: Breast milk lymphocytes

Dear Corrado, 
as you correctly identified, the presence of a large amount of fat in your
milk is what is causing problems. 
Some work has already been done, mainly in ruminants milk, but in humans as
well.
If you type:

milk flow cytometry

in PubMed you get 201 references where you can find some protocols to help
you.

Best regards

Mara

*******************************
Dr. Mara Rocchi DVM Ph.D. 
Flow Cytometer Manager	   
Moredun Research Institute
Pentland Science Park
Bush Loan, Penicuik
EH26 0PZ
Scotland, UK
Tel: +44-131-4455111
Fax: +44-131-4456111
e-mail: mara.rocchi@mri.sari.ac.uk


-----Original Message-----
From: Corrado Cilio [mailto:corrado.cilio@endo.mas.lu.se]
Sent: 20 January 2004 18:56
To: cyto-inbox
Subject: Breast milk lymphocytes


Dear flow community,

We are trying to run flow cytometry on different T cell populations in 
human breast milk . It is not easy to get rid of all the fat and although 
we succeeded in doing some phenotyping we are encountering two major
problems:
1. cell recovery after centrifugation and elimination of fat. If we count 
the cells before and after centrifugation and several washes we regularly 
experience great loss of cells.
2. When we stain for T cell markers as CD4 and CD8 we see to beautiful 
population but there is always a disturbing diagonal background which do 
not disappear no matter how strict we keep the lymphocyte gate on FCS/SSC.

There is someone out there that can give us some suggestions or have some 
more experience with breast milk lymphocytes?

Thank you in advance for any help and or advices.

Corrado

-------------------ooo--O..O--ooo----------------------------------
Corrado M. Cilio, M.D., Ph.D.
Dept. of Pediatrics and Dept. of Endocrinology
Malmoe University Hospital
Lund University
S-205 02 Malmoe, Sweden
Tel: +40-332395; mobil: 070-4330338; Fax: +40-337042
e-mail: corrado.cilio@endo.mas.lu.se
http://www.endo.mas.lu.se/html/w/wallen.html
  -------------------------------------------------------------- 

Judge a man by his questions rather than his answers.
  - Voltaire - 


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