Journal: Journal of visualized experiments : JoVE

Article Title: An Optimized O9-1/Hydrogel System for Studying Mechanical Signals in Neural Crest Cells

doi: 10.3791/62693

Figure Lengend Snippet: Materials

Article Snippet: It is better to determine which type suits the experiment best before starting the experiments (see the ). table ft1 table-wrap mode="anchored" t5 Name of Material/Equipment Company Catalog Number Comments/Description 12mm #1 Corning 0211 Glass Coverslip Chemglass Life Sciences CLS-1763-012 2% Bis-Acrylamide Sigma Aldrich M1533 24-well plate Greiner Bio-one 662165 25mm #1 Corning 0211 Glass Coverslip Chemglass Life Sciences CLS-1763-025 3-aminopropyl triethoxysilane (APTS) Sigma Aldrich A3648 4-well cell culture plate Thermo Scientific 179830 4% Paraformaldehyde Sigma Aldrich J61899-AP 40% Acrylamide Sigma Aldrich A4058 50% glutaraldehyde Sigma Aldrich G7651 6-well cell culture plate Greiner Bio-one 657160 AFM cantilever (spherical bead) Novascan AFM software Catalyst NanoScope Model: 8.15 SR3R1 Alexa Fluor 488 Phalloidin Thermo Fisher A12379 Ammonium Persulfate (APS) Sigma Aldrich 248614 Powder anti-AP-2α Antibody Santa Cruz sc-12726 anti-Vinculin antibody Abcam ab129002 Atomic Force Microscopy (AFM) Bioscope Catalyst Bruker Corporation Collagen type I (100mg) Corning 354236 DAPI (4’,6-Diamidino-2-Phenylindole, Dihydrochloride) Thermo Fisher D1306 Dichloromethylsilane (DCMS) Sigma Aldrich 440272 Donkey serum Sigma Aldrich D9663 Dulbecco’s Modified Eagle Medium (DMEM) Corning 10-017-CV Fetal bovine serum (FBS) Corning 35-010-CV Fluorescence microscope Leica Model DMi8 Fluoromount-G mounting medium SouthernBiotech 0100-35 HEPES Sigma Aldrich H3375 Powder Horse serum Corning 35-030-CI iScript Reverse Transcription Supermix Bio-Rad 1708841 Penicillin-Streptomycin antibiotic Thermo Fisher 15140148 RNeasy micro kit Qiagen 74004 Sterile 1x PBS Hyclone SH30256.02 Sterile deionized water Hardy Diagnostics U284 sulfo-SANPAH Thermo Fisher 22589 SYBR green Applied Biosystems 4472908 TEMED Sigma Aldrich T9281 Triton X-100 Sigma Aldrich X100 Tween 20 Sigma Aldrich P9416 Open in a separate window Materials Use an alcohol burner or Bunsen burner to sterilize each coverslip by passing it back and forth through the flame (30 s for protein assay experiments).

Techniques: Cell Culture, Software, Microscopy, Modification, Fluorescence, SYBR Green Assay