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- n. A vibrating microtome used to section soft tissue without freezing
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Results are presented in Coronal slices (100-µm thick) of rat brains were cut with a VT 1000S vibratome (Leica) and directly fixed for 1 h in iced 4% paraformaldehyde in phosphate-buffered solution (PBS).
Coronal slices were obtained by cutting with a Leica vibratome VT1000S in the presence of the ionotropic glutamate receptor inhibitor kynurenic acid (2 mM).
Immunohistochemistry on vibratome slices of paraformaldehyde-fixed tissue was performed using standard procedures as described Functional complementation of NCre and CCre in vitro.
For cross sections, fins were embedded in standard condition and section (20 µm) performed using a vibratome.
Retinas from 6-8 week old C57BL / 6J mice were dissected and flattened by making 4 radial cuts, then placed on a 20\% gelatin block secured to a vibratome chuck.
Photoreceptor cells collected by enzymatic digestion (papain) of vibratome-isolated photoreceptor sheets were cultured in 96 well laminin-coated plates (Sigma) at 37°C under 5\%
Fifty-micrometer horizontal vibratome sections were then cut, cryoprotected in 2\% glycerol and 20\% DMSO in 0.1 M PB for 20 min, and freeze-thawed eight times in liquid nitrogen.