Cleaning coverslip properly is crucial for imaging. I have collected several protocols amongst MBI labs. Each is used for slightly different purpose.
1. Nitric Acid Cleaning
Used by: Chew Lili and Edna,
For: Talin Stretching TIRF Single Molecule Imaging
a. soak coverslips in 10M Nitric Acid for 20 mins
b. wash the coverslips 8 times in MilliQ water by soaking them for 10 mins each
c. air dry the coverslips and keep them in a sealed container
Note: All action should be taken in bio-safety cabinet.
These Coverslips are coated with fibronectin before seeding.
2. Sulphuric acid Cleaning
Used by: Fu HongXia
For: Single DNA Strectching by Magnetic Tweezers
a. soak coverslips in acetone for 30 mins in sonicator
b. soak them in methanol for 30 mins in sonicator
c. rinse well with MilliQ water in sonicator
d. prepare acid wash solution:
Water should be added first.
Warning! Be extra careful when handling this acid.
e. soak the coverslip in the acid solution, slowly heat it up to around 100oC
f. rinse well with MilliQ water
g. clean these coverslips with plasma cleaning
Note: All actions should be taken in fume hood (except for plasma cleaning)