Re: Low level fluorescence

Deborah Berglund (umbbd@gemini.oscs.montana.edu)
Sat, 9 Dec 1995 14:48:48 -0700 (MST)

Try getting your antibody conjugated to biotin, then use the biotin
avidin linkage with a known number of fluorescein (or PE...) molecules.
Worked for us.
Deb from Bozeman, MT

On Thu, 7 Dec 1995, Susan & Ulrik Sprogxe-Jakobsen wrote:

> Hello out there!
>
> I am performing flow cytometry analysis on human red cells. Due to the low
> number of antigen copies per cell, the resulting fluorescense (FITC) is
> close to the flowcytometer (FACScan) noise level.
>
> 1. Does anyone know how to calculate the real number of cell-bound FITC
> molecules (and hence antigens), when working in this low area of
> fluorescence intensity?
>
> 2. Unmarked human red cells have a lower green (FITC) autofluorescence
> than certified blank beads. Any suggestions on how to incorporate this in
> the calculations of cell-bound FITC molecules on human RBCs?
>
>
> Any suggestion appreciated ...
>
> Best regards
>
> Ulrik Sprogoe-Jakobsen
>
> Dept. Clinical Immunology
> Odense University Hospital
> DK-5000 Odense C
> Phone: +45 6541 3576
> Fax: +45 6612 7975
>
>
>


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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone: (765)-494-0757; FAX(765) 494-0517; Web http://www.cyto.purdue.edu , EMAIL cdrom3@flowcyt.cyto.purdue.edu


Home Page Table of Contents Sponsors E-Mail Archive Web Sites

CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the Purdue University Cytometry Laboratories and distributed free of charge as an educational service to the cytometry community. If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL, Purdue University, West Lafayette, IN 47907. Phone: (765)-494-0757; FAX(765) 494-0517; Web http://www.cyto.purdue.edu , EMAIL cdrom3@flowcyt.cyto.purdue.edu