Subbing Coverglass for STORM tissue-sections


  • Thomas CoorsTek ceramic coverglass holders (2)
  • 1M KOH (56.11 g KOH in 1000 ml H2O)
  • 600 ml pyrex beakers (2)
  • Gelatin (from bovine skin type B)
  • Chromium potassium sulfate
  • 95% EtOH (ethanol)


  1. Place coverglass in ceramic holders: for STORM imaging, #1.5 (water immersion objective) or #1 (oil immersion objective) coverglass is appropriate
  2. Place racks with coverslips in 600 ml beaker
  3. Fill beaker with 1M KOH
  4. Place beaker in sonicator for 15 minutes
  5. Prepare gelatin

– 1,500 mg gelatin in 290 ml H2O. Heat to dissolve. (5x previous recipe)
– Prepare sodium potassium sulfate solution:
– 150 mg chromium potassium sulfate in 10 ml H2O. Vortex to dissolve. (5x previous recipe)
– Add chromium potassium sulfate solution to gelatin solution. Total volume is now 300 ml.
6. Rinse coverglass 1x in ddH2O
7. Dispose of KOH into appropriate waste container
8. Rinse out KOH with ddH2O
9. Fill beaker with 95% EtOH
10. Place racks with coverglass in EtOH-filled beaker
11. Return to sonicator for 15 minutes
12. Remove ceramic racks and let coverglass air dry. Use KimWipes to wick
away excess EtOH from the base of the coverglass and the troughs of the
ceramic holder.
13. Dip racks with coverglass into gelatin/chromium potassium sulfate solution
and swirl for 1 minute
14. Air dry coverglass. Use KimWipes to wick away excess solution from the
base of the coverglass and the troughs of the ceramic holder. Carefully
separate the coverslips from one another with a forceps (to ensure even
drying without aggregation of gelatin into smears or blobs).
15. Store coverglass for future use.

  • Adopted from Colenso Speer’s protocol
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