Onde cortar matérias: Uma dissecção e análise de guia para a orientação espacial da Retina do rato de Marcos Ocular
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Summary
Abstract
Introduction
Protocol
Representative Results
Discussion
Disclosures
The authors have nothing to disclose.
Acknowledgements
Materials
| 0.1 M Phosphate Buffered Saline | Sigma-Aldrich | P5244 | |
| Axioplan2 Epifluorescent Microscope | Zeiss | N/A | |
| Clear Nailpolish | N/A | N/A | |
| Corning LSE Low Speed Orbital Shaker | Sigma-Aldrich | CLS6780FP | |
| Costar TC-Treated 24-well Plates | Sigma-Aldrich | CLS3524 | |
| Dissection Microscope | Olympus | SZ51 | |
| Donkey anti-Goat Alexa 594 | Life Technologies | A11058 | |
| Donkey anti-Rabbit Alexa 594 | Life Technologies | A21207 | |
| Donkey Normal Serum | Millipore | 566460 | Use at 5.2% (52 μL with 86 μL of 20% Triton X-100 and 863 μL of 0.1M PBS for 1 mL of blocking solution) |
| Fisherbrand Superfrost Plus Microscope Slides | Fisher Scientific | 12-550-15 | |
| Goat anti-s-opsin | Santa Cruz Biotechnologies | sc-14363 | Not commerically available as of 2017 |
| Graefe Curved Forceps | Fine Science Tools | 11052-10 | |
| ImageJ or FIJI | National Institute of Health | N/A | Freely available software |
| Low Temperature Cautery Ophthalmic Fine Tip Cauterizer | Bovie Medical Corporation | AA00 | |
| MATLAB | MathWorks | N/A | At least version 2007b or later |
| Micro Cover Glasses | VWR International | 48393-241 | |
| Micro Slide Trays | VWR International | 82020-913 | |
| Moira Ultra Fine Forceps | Fine Science Tools | 11370-40 | |
| Nitrocellulose membrane | Millipore | HAWP04700 | |
| Paraformaldehyde | Electron Microscopy Sciences | 15714-S | Use at 4% (25 μL and 875 μL of 0.1 M PBS for 1 mL of fixative) |
| PrecisionGlide Needle 20G (0.90 mm x 25 mm) | BD PrecisionGlide | 305175 | |
| Pyrex Glass Petri Dish | Sigma-Aldrich | CLS3160152 | |
| R | The R Project for Statistical Computing | N/A | Freely available software; version 3.4.3 or later |
| Rabbit anti-s-opsin | Millipore | ABN1660 | |
| Retiga R3 Microscope Camera | Qimaging | 01-RET-R3-R-CLR-14-C | |
| Retistruct | N/A | N/A | Freely available software compatiable with Windows 7 or Windows 10 |
| Shandon Aqua-Mount Slide Mounting Media | Fisher Scientific | 14-390-5 | |
| Triton X-100 | Sigma-Aldrich | T8787 | Use 1.7% (86 μL of 20% Triton-X with 52 μL of Donkey Normal Serum and 863 μL of 0.1 M PBS for 1 mL of blocking solution) |
| Vannas Spring Dissection Scissors | Fine Science Tools | 15000-03 | |
| 5MP USB Microscope Digital Camera | AmScope | MU500 | To be used with the Olympus Dissection Microscope |
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Cite This Article
Sondereker, K. B., Stabio, M. E., Jamil, J. R., Tarchick, M. J., Renna, J. M. Where You Cut Matters: A Dissection and Analysis Guide for the Spatial Orientation of the Mouse Retina from Ocular Landmarks. J. Vis. Exp. (138), e57861, doi:10.3791/57861 (2018).