Bulk - cultured PC3 - GFP cells (A), holoclone P1H8 (B)-- , or meroclone P2H3 (C)-- derived cells were plated
on glass coverslips and stained for CD44, α2β1, and / or β - catenin using monoclonal antibodies.
Sections were mounted
with glass coverslips and images were taken using a microscope (Axioplan; Zeiss) connected to a CCD camera (RT slider; Diagnostic Instruments).
Bulk - cultured PC3 or PC3 - GFP cells, or clone - derived cultures were plated
on glass coverslips and used in immunofluorescence staining for CD44, α2β1 integrin, and β - catenin using monoclonal antibodies and protocols described previously (27 — 29).
Chambers were 500 μm wide, 100 μm high and 7 mm long, and were constructed from poly (dimethylsiloxane) bonded to
glass coverslips.
The cultures were then incubated at room temperature for one hour to allow cells to attach to
the glass coverslip.
Cells were plated either on
glass coverslips (40,000 cells / coverslip) coated with poly - L lysine and laminin or six - well plates (106 cells / well) coated with laminin alone.
Basically, the veterinarian will mix a fresh fecal sample from the dog with a special solution, strain the mixture, put it into a test tube covered with a glass lid, spin it down in a centrifuge and then look at the material that sticks to the underside of
the glass coverslip under a microscope.