Single Cell Dissociation of Caenorhabditis elegans: A Method to Isolate Live Cells
Published: April 30, 2023
Abstract
Source: Germany, E. M., et al. Isolation of Specific Neuron Populations from Roundworm Caenorhabditis elegans. J. Vis. Exp. (2019).
This video describes a method to dissociate whole worms into a single-cell suspension suitable for FACS or immunoprecipitation of intact cells.
Protocol
This protocol is an excerpt from Germany et al, Isolation of Specific Neuron Populations from Roundworm Caenorhabditis elegans, J. Vis. Exp. (2019). 1. Preparation and collection of aged worms for cell isolation NOTE: Explained below is the isolation of cholinergic neurons…
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).