| 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.
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| 6.
| Transfer slides to polylysine solution and shake 15 min. - 1 hr.
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| 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.
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|
| Transfer slide racks to empty chambers with covers for transport to vacuum oven.
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| 9.
| Dry slide racks in 45C vacuum oven for 10 min. (Vacuum is optional.)
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| 10.
| Store slides in closed slide box (plastic only, without rubber mat bottom)
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| 11.
| BEFORE PRINTING ARRAYS:
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|
| Check that polylysine coating is not opaque.
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|
| Test print, hyb and scan sample slides to determine slide batch quality. |
