Enkelt celle dissociation af Caenorhabditis elegans:En metode til at isolere levende celler
Published: April 30, 2023
Abstract
Kilde: Tyskland, E.M., et al. Isolering af specifikke Neuron Populationer fra Roundworm Caenorhabditis elegans. J. Vis. Exp. (2019).
Denne video beskriver en metode til at adskille hele orme i en enkelt celle suspension egnet til FACS eller immunoprecipitation af intakte celler.
Protocol
Denne protokol er et uddrag fra Tyskland et al, Isolation af specifikke neuronpopulationer fra Roundworm Caenorhabditis elegans, J. Vis. Exp. (2019). 1. Forberedelse og indsamling af lagrede orme til celleisolation BEMÆRK: Forklaret nedenfor er isoleringen af cholinrgiske neuroner…
Materials
| Agar, Molecular Biology Grade | VWR | A0930 | |
| CaCl2 | Sigma | C5670 | |
| Contess Automated Cell Counter | Invitrogen | Z359629 | |
| DTT | USBiological | D8070 | |
| FBS | Omega Scientific | FB-02 | |
| Fluorodeoxyuridine | Sigma | F0503 | |
| HEPES | Sigma | H3375 | |
| KCl | Amresco | O395 | |
| KH2PO4 | USBiological | P5110 | |
| Leibovitz's L-15 Medium | Gibco | 21083027 | |
| MgCl2 | Sigma | M8266 | |
| MgSO4 | USBiological | M2090 | |
| Na2HPO4·7H2O | USBiological | S5199 | |
| NaCl | VWR | X190 | |
| NaOCl (Bleach) | Clorox | ||
| Penicillin-Streptomicen | Fisher Scientific | 15140122 | |
| Pronase E | Sigma | 7433 | protease mixture from Streptomyces griseus |
| SDS | Amresco | O227 | |
| SMT1-FLQC fluorescence stereomicroscope | Tritech Research | ||
| Sucrose | USBiological | S8010 |
Cite This Article
Single Cell Dissociation of Caenorhabditis elegans: A Method to Isolate Live Cells. J. Vis. Exp. (Pending Publication), e20135, doi: (2023).