Freeze-Cracking of Nematodes: A Method to Expose Interior Worm Tissues for Staining
Published: April 30, 2023
Abstract
Source: 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. Exp. (2017).
This video describes freeze-cracking, a method to make C. elegans tissues accessible for antibody staining by disrupting the cuticle.
Protocol
This protocol is an excerpt from 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). Antibody staining of the C. elegans intestine Fixation Take a clean glass slide and use poly-L-lysine to genera…
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).