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Preparation of Slides
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PreparationofSlides
Lastupdated:October6,1999Materials Qty Orderinfo Glassmicroscopeslides 60 GoldSeal#3010 Sliderack 2 ShandonLipshaw#121(800-245-6212) <=Eachrackholds30slides Slidechamber 6 ShandonLipshaw#121 <=Eachchamberholds350mL ddH2O ~5L NaOH 70g 95%Ethanol 420mL Poly-L-lysine 70mL Sigma#P8920 TissueculturePBS 70mL Vacuumoven(45C) Slidebox(plasticonly) 1 VWR#48443-806 1. Placeslidesinslideracks.Placeracksinchambers. 2. PrepareCLEANINGSOLUTION: Dissolve70gNaOHin280mLddH2O. Add420mL95%ethanol.Totalvolumeis700mL(=2X350mL);stiruntilcompletelymixed. Ifsolutionremainscloudy,addddH2Ountilclear. 3. Poursolutionintochamberswithslides;coverchamberswithglasslids.Mixonorbitalshakerfor2hr. Onceslidesareclean,theyshouldbeexposedtoairaslittleaspossIBLe.Dustparticleswillinterferewithcoatingandprinting. 4. QuicklytransferrackstofreshchambersfilledwithddH2O.Rinsevigorouslybyplungingracksupanddown. Repeatrinses4XwithfreshddH2Oeachtime.ItiscriticaltoremovealltracesofNaOH-ethanol. 5. PreparePOLYLYSINESOLUTION: 70mLpoly-L-lysine+70mLtissueculturePBSin560mLwater. Useplasticgraduatedcylinderandbeaker. 6. Transferslidestopolylysinesolutionandshake15min.-1hr. 7. TransferracktofreshchambersfilledwithddH2O.Plungeupanddown5Xtorinse. 8. Centrifugeslidesonmicrotiterplatecarriers(placepapertowelsbelowracktoabsorbliquid)for5min.@500rpm. Transfersliderackstoemptychamberswithcoversfortransporttovacuumoven. 9. Dryslideracksin45Cvacuumovenfor10min.(Vacuumisoptional.) 10. Storeslidesinclosedslidebox(plasticonly,withoutrubbermatbottom) 11. BEFOREPRINTINGARRAYS: Checkthatpolylysinecoatingisnotopaque. Testprint,hybandscansampleslidestodetermineslidebatchquality.
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