Preparation of Slides
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| Materials | Qty | Order info | |
|---|---|---|---|
| Glass microscope slides | 60 | Gold Seal #3010 | |
| Slide rack | 2 | Shandon Lipshaw #121 (800-245-6212) | <= Each rack holds 30 slides |
| Slide chamber | 6 | Shandon Lipshaw #121 | <= Each chamber holds 350 mL |
| ddH2O | ~5 L | ||
| NaOH | 70 g | ||
| 95% Ethanol | 420 mL | ||
| Poly-L-lysine | 70 mL | Sigma #P 8920 | |
| Tissue culture PBS | 70 mL | ||
| Vacuum oven (45C) | |||
| Slide box (plastic only) | 1 | VWR #48443-806 |
| 1. | Place slides in slide racks. Place racks in chambers. |
| 2. | Prepare CLEANING SOLUTION: |
| Dissolve 70 g NaOH in 280 mL ddH2O. | |
| Add 420 mL 95% ethanol. Total volume is 700 mL (= 2 X 350 mL); stir until completely mixed. | |
| If solution remains cloudy, add ddH2O until clear. | |
| 3. | Pour solution into chambers with slides; cover chambers with glass lids. Mix on orbital shaker for 2 hr. |
| Once slides are clean, they should be exposed to air as little as possible. Dust particles will interfere with coating and printing. | |
| 4. | Quickly transfer racks to fresh chambers filled with ddH2O. Rinse vigorously by plunging racks up and down. |
| Repeat rinses 4X with fresh ddH2O each time . It is critical to remove all traces of NaOH-ethanol. | |
| 5. | Prepare POLYLYSINE SOLUTION: |
| 70 mL poly-L-lysine + 70 mL tissue culture PBS in 560 mL water. | |
| Use plastic graduated cylinder and beaker. | |
| 6. | Transfer slides to polylysine solution and shake 15 min. - 1 hr. |
| 7. | Transfer rack to fresh chambers filled with ddH2O. Plunge up and down 5X to rinse. |
| 8. | Centrifuge slides on microtiter plate carriers (place paper towels below rack to absorb liquid) for 5 min. @ 500 rpm. |
| Transfer slide racks to empty chambers with covers for transport to vacuum oven. | |
| 9. | Dry slide racks in 45C vacuum oven for 10 min. (Vacuum is optional.) |
| 10. | Store slides in closed slide box (plastic only, without rubber mat bottom) |
| 11. | BEFORE PRINTING ARRAYS: |
| Check that polylysine coating is not opaque. | |
| Test print, hyb and scan sample slides to determine slide batch quality. |
