How to sterilize cover slips (glasses)? - (Jan/30/2009 )
We use coverslips for cell/tissue culture to prepare samples for immunohistochemical analysis.
I do this under a hood with a lamina flow. i put the slides in 96%etoh for some minutes, then flame them and let them dry.
afterwards i put them in poly-l-lysin for some minutes, then I aspirate the excess of poly-l-lysin and thats it.
the poly-l-lysin is not for necessary for sterilization, but several cell-lines need it to attacht to the slides
We also autoclave our coverslips. We are using Fisherbrand 18 mm round coverslips. We individually wrap 1 to 3 coverslips together, depending on the number needed, in aluminum foil. Wrap neatly and very carefully, so they are easy to remove with tweezers later. We autoclave them on the dry cycle for about 30 min and they survive perfectly. For a different brand though, I would recommend testing 1 or 2 first. Put them in an erlenmeyer flask, or pipette tip box, so they are protected during and after autoclaving. You can then store them this way for several weeks following autoclaving. For added sterilization, we flame them one by one as they are taken out of the aluminum foil via tweezers, and then place them in the wells of the plate. One or two passes through the flame, without ethanol, is fine. We grow cells on them for up to 5 days in various treatments and conditions and do not have problems with contamination. We also have not had problems with contamination when they are not flamed, but now used it as an added security.
No need to sterilize them, just open a new box of cover slips in hood and use.
I never get contamination. I use corning cover slips.
In my lab we put c/s overnight in nitric acid, on the other day we wash 3 times during 30' with water. Then 30' in 100% ethanol and 3 more washes with water. They are kept in water at RT.
I used to put coverslips one by one on the "Tip BOX". After filling one layer, I put a tissue paper cover them. Again, placed coverslips one by one. You could do several rounds till amount you need. Then autoclaved the "Tip Box".
I wash rinse them with distilled water for 10 mins
Then wash them with 90% alcohol for 15 mins
Wash them again with distilled water
Autoclave by placing the bunch in a beaker sealed with aluminium foil.
air dry in the warm room overnight.
I actually read this another procedure from a book which involves detergents. I will send you the procedure once I find it.
But my procedure has been working well so far
I moved to a new lab and have to set-up things for IF. Can any of you guys tell me the vendor and catalog number of coverslips that fit-in to a 24-well plate, as well as the standard glass slides with frosted-end?
Thank you in advance,
We normally autoclave them and after dry them in the heater.
The problem is that if they are superimposed they probably will be harnessed, so it is important to sort them in order to maintain them separated.
We use tips boxes because they have holes and it is easier to separate the cover slips from the surface
labrat612 on Feb 4 2009, 12:44 PM said:
wash them in surfactant to remove grease, 70% ethanol and flame. The UV is silly- it's pretty good at blocking.