[Histonet] Alternatives to Cytospin?

[Tony Henwood]

Jerry,

Firstly, I would not recommend fixing in ethanol since cells will shrink and Romanowsky staining will be compromised (the cells stain too darkly since they are smaller).

I would use a centrifuge to concentrate the cells and smear like a blood film or if you do not have a centrifuge available, place a drop of the BALF on the slide and either air dry as you suggested (if cell count is low) or treat it like blood (if it looks quite opaque - lots of cells/ & other junk).

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of JR R
Sent: Friday, 3 June 2011 9:54 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Alternatives to Cytospin?


I will be collecting bronchoalveolar lavage fluid (BALF) from mice and want to do do a diff-quick stain on the fluid to characterize the leukocyte population.

My colleagues usually use a cytospin to concentrate cells onto a slide but I'll be at a lab that doesn't have the equipment so I'm looking for alternatives.

Could I just put a drop of the BALF onto a charged slide and air dry it?  Or treat the BALF like a blood smear?

Alternatively, could I fix the cells in...ethanol maybe and then cytospin them the next day when I get back to my lab?

Any ideas will be much appreciated.

Jerry Ricks
Research Scientist
University of Washington
Department of Pathology


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