Freeze-Cracking av Nematoder: En metode for å eksponere innvendig ormvev for farging

Abstract

Kilde: Zhang, N., et al. The C. elegans Intestine As a Model for Intercellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis at the Single-cell Level: Labeling by Antibody Staining, RNAi Loss-of-function Analysis and Imaging. J. Vis. Uttr. (2017).

Denne videoen beskriver fryse-cracking, en metode for å gjøre C. elegans vev tilgjengelig for antistofffarging ved å forstyrre kutiklet.

Protocol

Denne protokollen er et utdrag fra Zhang et al, The C. elegans Intestine As a Model for Intercellular Lumen Morphogenesis and In Vivo Polarized Membrane Biogenesis at the Single-cell Level: Labeling by Antibody Staining, RNAi Loss-of-function Analysis and Imaging, J. Vis. Exp. (2017). Antistofffarging av C. elegans tarmen Fiksering Ta en ren glasssklie og bruk poly-L-lysin for å generere en tyn…

Materials

Antibody staining
poly-L-lysine	Sigma	P5899
Methanol	Fisher Scientific	A452-4
Acetone	Fisher Scientific	A949SK-4
Permount	Fisher Scientific	SP15-100
Microscope slides	Fisher Scientific	4448
Microscope coverslips (22×22-1)	Fisher Scientific	12-542-B
M9 Medium			lab made
OP50 bacteria	CGC