Analysis of surface properties of fixed and live cells using derivatized agarose beads.
A novel assay has been developed for the histochemical characterization of surface properties of cells based on their adhesion to agarose beads derivatized with more than 100 types of molecules, including sugars, lectins and other proteins, and amino acids. The assay simply involves mixing small quantities of washed cells and beads in droplets on glass microscope slides and determining to which beads various cell types adhere. Distilled water was found to be the best medium for this assay because added ions or molecules in other media inhibit adhesion in some cases. Many cells, however, cannot tolerate distilled water. Here we show that cells fixed with either of two fixatives (1% formaldehyde or Prefer fixative) displayed similar bead-binding properties as did live cells. Specificity of cell-bead binding was tested by including specific free molecules in the test suspensions in hapten-type inhibition experiments. If a hapten compound inhibited live-cell adhesion to a specific bead, it also inhibited fixed-cell adhesion to a specific bead. The results of these experiments suggest that fixed cells display authentic surface properties, opening the door for the use of this assay with many cell types that cannot tolerate distilled water.
Navarro
Vanessa M
VM
Department of Biology and Center for Cancer and Developmental Biology, California State University, Northridge 91330-8303, USA.
Walker
Sherri L
SL
Badali
Oliver
O
Abundis
Maria I
MI
Ngo
Lylla L
LL
Weerasinghe
Gayani
G
Barajas
Marcela
M
Zem
Gregory
G
Oppenheimer
Steven B
SB
D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism*; D006241:Haptens / Q000276:immunology; D008863:Microspheres; D011956:Receptors, Cell Surface / Q000276:immunology / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D012680:Sensitivity and Specificity; D012685:Sepharose / Q000276:immunology / Q000378:metabolism*; D013499:Surface Properties; D016707:Tissue Fixation
D006241:Haptens; D011956:Receptors, Cell Surface; D012685:Sepharose