We use various sorts of flowcells.  These can be as simple as a plexiglass
block with a couple of tracks cut in it and a coverslip glued on top to
seal things up (see Palmer, Meth Enzymol. V310).  We have not used
eukaryotic tissue culture cells (the cells to which you are referring I
presume), but this approach works fine for bacteria and yeasts.  They atach
nicely and don't care which way gravity is going...however, non-motile
cells need to be seeded in the "upside down" position (so they fall down
onto the coverslip) before turning the flowcell over and beginning
perfusion.


>Hi,
>
>Is anyone using an upright fluorescence microscope to analyze viable cells
>in a closed culture system with a 100x oil objective. i.e., the cells will
>have to be upside down on the upper cover glass and the chamber filled with
>culture fluid?
>
>If so, did you have to make your culture holder or are there commercially
>available systems that will accomplish this?  The Bioptechs catalog shows
>the FCS3 which would permit this but I'm not sure if it's in production yet.
>
>Thanks for any help.
>
>Ray
>
>Raymond B. Hester, Ph.D.
>Research Cytometry Laboratory
>CSAB 357, Biotechnical Services
>University of South Alabama
>Mobile, AL 36688
>Voice:    (334) 460-6029
>FAX:      (334) 460-6073
>E-mail:   [log in to unmask]

Robert J. Palmer Jr., Ph.D.
Natl Inst Dental Craniofacial Res - Natl Insts Health
Oral Infection and Immunity Branch
Bldg 30, Room 308
30 Convent Drive
Bethesda MD 20892
ph 301-594-0025
fax 301-402-0396