Congelación de nematodos: un método para exponer los tejidos de gusano interiores para la tinción
Published: April 30, 2023
Abstract
Fuente: 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 Anticuerpo Staining, RNAi Loss-of-function Analysis and Imaging. J. Vis. Exp. (2017).
Este video describe el liongelante, un método para hacer que los tejidos C. elegans sean accesibles para la tinción de anticuerpos alterando la cutícula.
Protocol
Este protocolo es un extracto de 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). Tinción de anticuerpos del intestino C. elegans fijación Tome una diapositiva de vidrio limpia y use poli-L-lisi…
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 |
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Freeze-Cracking of Nematodes: A Method to Expose Interior Worm Tissues for Staining. J. Vis. Exp. (Pending Publication), e20130, doi: (2023).