Daniel:Protocols/NuclearAdhesion
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Nuclear Adhesion to Slides[edit]
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