Daniel:Protocols/NuclearAdhesion

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Nuclear Adhesion to Slides[edit]

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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.

  1. Clean glass slide in 100% EtOH; allow to air dry
    1. Draw a square around the desired adhesion area using a hydrophobic barrier pen
  2. Remove cells from -80C and spin down for 5 min at 1000g
    1. Remove supernatant, careful not to disturb the pellet
  3. Add 500uL of 4% paraformaldehyde to the tube and wait 5 min
  4. Centrifuge cells for 5 min at 1000g
    1. Remove supernatant, careful not to disturb the pellet
  5. Add 500 uL of 70% EtOH
  6. Centrifuge cells for 5 min at 1000g
    1. Remove supernatant, careful not to disturb the pellet
  7. Resuspend nuclei in 20 uL 70% EtOH
  8. Deposit on to glass slide
  9. Use a coverslip to gently spread the 20 uL across the slide
    1. Be careful not to scratch the slide
  10. Dry the slide using a heating block at 60C for 10-15 min