100 mm culture dish |
Corning |
353025 |
Used for dissection (steps 1.3, 3 and 5) |
50 mL tube |
Genesee Scientific |
28–106 |
For fixation and cryoprotection (step 1) |
Cryostat |
Leica |
CM1850 |
For cryosectioning (step 7) |
Curved scissors |
Fine Science Tools |
91500-09 |
Used for dissection (steps 1.3, 3 and 5) |
DAPI |
Fisher Scientific |
D3571 |
Diluted 1:1,000 in blocking buffer |
Dissection microscope |
Zeiss |
Stemi 2000-C |
Used for dissection (steps 1.3, 3 and 5) |
Donkey anti-Goat 488 |
Fisher Scientific |
A-11055 |
Diluted 1:1,000 in blocking buffer |
Donkey anti-Mouse 555 |
Fisher Scientific |
A-31570 |
Diluted 1:1,000 in blocking buffer |
Forceps |
Fine Science Tools |
91150-20 |
Used for dissection (steps 1.3, 3 and 5) |
Glass Slide Cover |
VWR |
48404-453 |
For cryosectioning (step 7) |
Goat anti-SOX2 |
R&D Systems |
AF2018 |
Diluted 1:100 in blocking buffer |
High-profile disposable cryostat blades |
Leica Microsystems Inc. |
14035838926 |
For cryosectioning (step 7) |
Kimwipe |
Fisher Scientific |
06-666-A |
Used to wipe away excess PBS or OCT (steps 3 and 6) |
Mouse anti-RPE65 |
Novus Bio |
NB100-355SS |
Diluted 1:100 in blocking buffer |
OmniPur Sucrose |
Millipore |
167117 |
Used for cryoprotectant (step 1.2) |
Paraformaldehyde 20% solution |
Electron Microscopy Sciences |
15713 |
Used as tissue fixative (diluted to 4% in step 1.1) |
Peel-A-Away Disposable Embedding Mold (22x22x20 mm Deep) |
Polysciences, Inc. |
18646A |
Used as embedding mold (step 6) |
Phosphate buffered saline, 1x |
Corning |
21-030-CV |
Used in preparation of fixative (step 1.1) and cryoprotectant (step 1.2) |
Scalpel blade no. 15 |
Feather |
08-916-5D |
Used for dissection (steps 1.3, 3 and 5) |
Superfrost Plus Microscope Slides |
Fisher Scientific |
12-550-15 |
For cryosectioning (step 7) |
Tissue-Tek O.C.T. Compound |
Sakura |
4583 |
Used as embedding media (step 6) |