Re: Dyes for cell tracking experiments...

Terry Hoy (WHGTH@cardiff.ac.uk)
Wed, 24 Jul 1996 09:55:43 GMT

>So, the question is this: which dye would any of you esteemed flow
>gurus recommend for this, which will:-
>
> Readily be taken up by viable cells.
> Not be removed by washing immediately afterwards.
> Persist for at least two hours, without serious loss of fluorescence.
>

Zip,
We use biotin to label red cells for in vivo tracking which
you may find suitable.
Labelling is simple : 0.3pg biotin / (red) cell. NHS-LC-Biotin,
Pierce,Rockford,IL in 0.9%NaCl for 10 mins at room temperature.
Wash twice before use.
It is long lived on RBC's - we have seen it many weeks later after
reinjection in vivo.
Detection with streptavidin-FITC gives a good signal to noise ratio -
we detect 1 in 1000 easily with 3% precision.
If you have sufficient material and FACStime going down to 1 in 100000
should be possible.
Details in "Red Cells and Reticulocytes" Chapter 8 ,Flow Cytometry :
Clinical Applications. Ed M.Macey (Blackwell).
Regards,
Terry.

Dr. Terry Hoy
Principal Research Officer
Department of Haematology Telephone 44 1222 743458
University of Wales College of Medicine FAX 44 1222 744655
Heath Park E-mail hoy@cardiff.ac.uk
CARDIFF CF4 4XN http://www.cf.ac.uk/uwcm/hg/hoy/index.html
U.K.