Cell-spreading assays.
A method is provided to quantitate the extent of cell spreading as a function of the expression level of transfected recombinant proteins. This chapter contains protocols for 1) replating and staining transfected cells for immunofluorescence microscopy, 2) optimizing image acquisition so that fluorescence intensity can be measured independent of cell morphology, and 3) quantitating cell area and expression levels of recombinant proteins for individual transfected cells using ImagePro-Plus software. This method can be used to further our understanding of intracellular signals and protein interactions that regulate cell spreading.
Berrier
Allison L
AL
LSU School of Dentistry, Department of Cell Biology and Anatomy, New Orleans, LA, USA.
LaFlamme
Susan E
SE
D000595:Amino Acid Sequence; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D016650:Fluorescein-5-isothiocyanate; D005455:Fluorescent Antibody Technique; D006801:Humans; D007231:Infant, Newborn; D039641:Integrin beta Chains / Q000737:chemistry / Q000502:physiology; D008297:Male; D008969:Molecular Sequence Data; D012867:Skin / Q000166:cytology; D012984:Software; D014162:Transfection
D039641:Integrin beta Chains; D016650:Fluorescein-5-isothiocyanate