C. 예인대 Blastomere 해부 : 달걀 껍질을 제거하고 배아 세포를 해리하는 방법
Published: April 30, 2023
Abstract
출처: Hsu, C. R., 외. 고립 된 Caenorhabditis elegans 배아 Blastomeres 및 접착 폴리스티렌 구슬과 공간 세포 접촉 패턴의 시험관 재구성. J. 비스 익스펙. (2019).
이 비디오는 초기 배아에서 C. elegans blastomeres를 격리하는 방법을 소개합니다. 결과 세포는 세포 배양 또는 전 생체 내 실험에 적합합니다.
Protocol
이 프로토콜은 Hsu 외에서 발췌, 격리 된 Caenorhabditis elegans 배아 Blastomeres 및 접착 폴리스티렌 구슬, J. Vis. Exp. (2019)와 공간 세포 접촉 패턴의생체로 재구성이다. 배아 블라도미어의 격리 장갑과 실험실 코트를 착용하여 절단및 표백 용액과 접촉하?…
Materials
| Aspirator Tube Assembly | Drummond | 21-180-13 | For the blastomere isolation. |
| Caenorhabditis elegans strain: N2, wild-type | Caenorhabditis Genetics Center | N2 | Strain used in this study |
| Caenorhabditis elegans strain: KSG5, genotype: zuIs45; itIs37 | in house | KSG5 | Strain used in this study |
| Calibrated Mircopipets, 10 µL | Drummond | 21-180-13 | For the blastomere isolation |
| CD Lipid Concentrate | Life Technologies | 11905031 | For the blastomere isolation. Work in the tissue culture hood. |
| Clorox | Clorox | N. A. | For the blastomere isolation. Open a new bottle when the hypochlorite treatment does not work well. |
| Coverslip holder | In house | N.A. | For the blastomere isolation. |
| Dissecting microscope: Zeiss Stemi 508 with M stand. Source of light is built-in LED. Magnification of eye piece is 10X. | Carl Zeiss | Stemi 508 | For the blastomere isolation. |
| Fetal Bovine Serum, Qualified One Shot, Canada origin | Gibco | A3160701 | For the blastomere isolation. Work in the tissue culture hood. |
| General Use and Precision Glide Hypodermic Needles, 25 gauge | BD | 14-826AA | For the blastomere isolation |
| Inulin | Alfa Aesar | AAA1842509 | For the blastomere isolation |
| MEM Vitamin Solution (100x) | Gibco | 11120052 | For the blastomere isolation. |
| Multitest Slide 10 Well | MP Biomedicals | ICN6041805 | For the blastomere isolation |
| Penicillin-Streptomycin (10,000 U/mL) | Gibco | 15140148 | For the blastomere isolation. |
| Polyvinylpyrrolidone | Fisher BioReagents | BP431-100 | For the blastomere isolation |
| Potassium Chloride | Bioshop | POC888 | For the blastomere isolation |
| Schneider’s Drosophila Sterile Medium | Gibco | 21720024 | For the blastomere isolation. Work in the tissue culture hood. |
| Sodium Chloride | Bioshop | SOD001 | For the blastomere isolation |
| Sodium Hydroxide Solution, 10 N | Fisher Chemical | SS255-1 | For the blastomere isolation |
| Syringe Filters, PTFE, Non-Sterile | Basix | 13100115 | For the blastomere isolation. |
| Tygon S3 Laboratory Tubing,, Formulation E-3603, Inner diameter 3.175 mm | Saint Gobain Performance Plastics | 89403-862 | For the blastomere isolation. |
| Tygon S3 Laboratory Tubing,, Formulation E-3603, Inner diameter 6.35 mm | Saint Gobain Performance Plastics | 89403-854 | For the blastomere isolation. |
Cite This Article
C. elegans Blastomere Dissection: A Method to Remove the Eggshell and Dissociate Embryonic Cells. J. Vis. Exp. (Pending Publication), e20158, doi: (2023).