Freeze-Cracking of Nematodes: A Method to Expose Interior Worm Tissues for Staining
Published: April 30, 2023
Abstract
Source : Zhang, N., et coll. 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).
Cette vidéo décrit le gel-fissuration, une méthode pour rendre les tissus C. elegans accessibles pour la coloration des anticorps en perturbant la cuticule.
Protocol
Ce protocole est un extrait de Zhang et coll.,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). Coloration d’anticorps de l’intestin de C. elegans fixation Prenez une lame de verre propre et utilisez la pol…
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 |
Citer Cet Article
Freeze-Cracking of Nematodes: A Method to Expose Interior Worm Tissues for Staining. J. Vis. Exp. (Pending Publication), e20130, doi: (2023).