इस अनुच्छेद के एक दृष्टिकोण के साथ और बिना रेटिना वर्णक उपकला एक से तीन दिन postfertilization भ्रूण से जुड़ी है, zebrafish retinas microdissect का वर्णन करता है.
Zebrafish आँख के ऊतकों के Microdissection प्रभावी ढंग से बरकरार retinas और RPE-संलग्न retinas प्राप्त कर सकते हैं. यह काफी हद तक अभिव्यक्ति एक विशिष्ट नेत्र ऊतक (यानी रेटिना या RPE) से संबंधित अध्ययन में मदद करता है. वास्तव में, हम सफल?…
The authors have nothing to disclose.
यह काम पर्ड्यू विश्वविद्यालय में एक जीव विज्ञान के विभाग से स्टार्टअप कोष द्वारा समर्थित है.
Material Name | Tipo | Company | Catalogue Number | Comment |
---|---|---|---|---|
Cordless pestle motor | VWR | 47747-370 | ||
DC power supply | Lascar | PSU130 | Any DC supply would work. The specific voltage of a different machine will need further optimization. | |
Disposable pestle & microtube, 1.5 mL (DNase, RNase and pyrogen-free) | VWR | 47747-366 | These are used for tissue collection in TRIzol for expression analysis. | |
Dumont #5 forceps, Tips: 0.05 x 0.01mm, Inox | World Precision Instruments | 500341 | Fine tip dimension is desirable but is not inflexible, as one may need to sharpen the tip from time to time. | |
Dumont #5SF forceps, Tips: 0.025 x 0.005mm, Inox | Fine Science Tools | 11252-00 | Fine tip dimension is desirable but is not inflexible, as one may need to sharpen the tip from time to time. | |
Falcon polystyrene culture plates, 60 X 15 mm | BD Biosciences | 351007 | These plates are used as dissection plates. | |
Olympus SZX16 Stereomicroscope | Olympus | SZX16 | Any stereomicroscope would work. We used Leica stereomicroscope in previous studies1-3 without any issues. We also use the 1X objective exclusively for the dissection even though we have a 2X objective installed. | |
Sharpening stone | Fine Science Tools | 29008-01 | Use this to sharpen the tip of the forceps if necessary | |
Thermo plate | Tokai Hit | MATS-U55SZX2B | This is used to maintain the temperature of the tissue throughout dissection and minimize the influence of temperature fluctuation on gene expression. We also put the whole microscope in an environmentally controlled room at 28°C during dissection in previous studies1-3 with good success. | |
Trizol, 100 mL | Invitrogen | 15596-026 | ||
tungsten wire, 0.015 inch diameter | World Precision Instruments | TGW1510 | ||
Wooden Applicator | Puritan | 807 | This is used for holding the chemically-etched tungsten needle. |