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=Nuclear Adhesion to Slides= [[Daniel Jacobsen|Back to Main]] This protocol is for the adhesion of nuclei on to glass slides for imaging. I have done it successfully on glass slides with poly-L lysine, not sure about Vecta-bond or uncoated. #Clean glass slide in 100% EtOH; allow to air dry ##Draw a square around the desired adhesion area using a hydrophobic barrier pen #Remove cells from -80C and spin down for 5 min at 1000g ##Remove supernatant, careful not to disturb the pellet #Add 500uL of 4% paraformaldehyde to the tube and wait '''5 min''' #Centrifuge cells for 5 min at 1000g ##Remove supernatant, careful not to disturb the pellet #Add 500 uL of 70% EtOH #Centrifuge cells for 5 min at 1000g ##Remove supernatant, careful not to disturb the pellet #Resuspend nuclei in 20 uL 70% EtOH #Deposit on to glass slide #Use a coverslip to gently spread the 20 uL across the slide ##Be careful not to scratch the slide #Dry the slide using a heating block at 60C for 10-15 min
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