Table 1:
Equipment and Reagents. List of equipment and reagents used in this study.
| Equipment |
| 50mL tube rotator |
| Orbital shaker |
| Vibratome (Leica VT1200s) |
| *Vibratome blade (TED Pella, 121-9) |
| *Scalpel or surgical scissors |
| Sharps waste container |
| Confocal microscope (Nikon Al R) |
| PC with image analysis software (ImageJ/FIJI, Nikon elements, or BitPlane Imaris) |
| *Hot plate |
| Paint brush, soft, synthetic bristles, #0. |
| Blunt Forceps |
| Luer Lok syringe (optional, for perfusion) |
| Blunt canula (optional, for perfusion) |
| Canula clip (optional, for perfusion) |
| Suture (optional, for perfusion) |
| Cryomold |
| Lab tissue paper (i.e. Kimwipes) or gauss |
| Cotton swab |
| Nitrite gloves |
| Polypropylene tubes (1.5 mL, 5 mL) |
| Nylon Mesh or Cell Strainer |
| 24 well plates |
| Falcon tubes (50 mL) |
| Sample baskets (5 mL tube + cell strainers/mesh) |
| Petri dishes |
| Coverslips (#0) |
| 100 mL Glass Jars |
| Coverslip caddy |
| Slides |
| Adhesive, solvent-resistant foil (Nashua Foil Tape, 279 pm thickness, catalog # 1207802) |
| Reagents |
| Phosphate buffered saline (PBS) (Gibco catalog # 21300-58) |
| KB buffer (mmol/L: KCI 20, KH2PO4 10, K+-glutamate 70, MgCl2 1, glucose 25, taurine 20, EGTA 0.5, HEPES 10, 0.1% bovine serum albumin, pH adjusted to 7.4 with KOH) |
| *16% paraformaldehyde, ampoule in nitrogen. (Alfa Aesar catalog # 43368) |
| *Methanol (Sigma catalog # 34860) |
| Low melt agarose (Fisher Scientific catalog # BP165-25) |
| Bovine Serum Albumin (BSA) |
| *Dimethyl Sulfoxide (DMS0) |
| *Sodium Azide |
| *Super glue |
| Normal Donkey Serum |
| *Triton X-100 |
| Fluorescent beads (Fluorsbrite, Thermofisher, Sigma) |
| Primary antibodies: phosphorylated serine 10 histone H3 (pH3, Abcam ab5176), trimethyl lysine 9 histone H3 (H3K9me3, Active Motif 39161), Heterochromatin Protein 1 gamma (HP1g, Active Motif 39981), Connexin 43 (Cx43) (Zymed catalog # 71-0700), GFP (Novus catalog # NB1001770). |
| Secondary antibodies: (Alexa Fluor 488, 555, and 647, Thermo Fisher) |
| Chemical dyes: Wheat Germ Agglutinin (WGA) Alexa Fluor-647, WGA-Oregon Green, phalloidin-Alexa Fluor 647, phalloidin-rhodamine, Hoechst 33342 |
| Poly-L-lysine |
| *lsopropanol |
| *Benzyl alcohol |
| *Benzyl benzoate |
| Nail polish (Sally Hanson lnsta-dry, clear) |
*
indicates hazardous reagent or equipment, please consult user manuals and MSDS for appropriate usage.