Likewise:
We remove the coverslip by soaking in 2X SSC [probably for no real reason,
but it helps preserve the specimen on the slide, and I suppose you could add
a drop of Tween 20 to the Coplin jar]. Recover and then rinse the coverslip
in deionised water, and finally wipe the coverslip carefully with ether or
petroleum spirit in the fume cupboard [gloves + KimTech Science 75512
tissues] to get it as clean as possible before the wash sequence below. We
use immersion oil for imaging [which doesn't dissolve so well in ethanol,
hence ether/petroleum spirit], and more importantly 'always liquid'
VectaShield + DAPI non-hardening mountant [naturally with no nail polish
gluing it on]. You can generally slowly soak off 'permanently' mounted
coverslips with things like Xylene/Toluene [check what the original mountant
was dissolved in, e.g. Histomount uses toluene]. After ether/petroleum
spirit cleaning, quickly visually inspect the cover-slips at an angle to the
reflected light to ensure they are smear free before the wash sequence
below.
[This wash sequence is also used for new 'pre-washed' glass slides from
boxes]: Soak the cleaned coverslips overnight in 5ml Teepol/RenClean in ~500
ml [try a slotted rack to hold the coverslips upright]. Wash off detergent
gently with tap water then de-ionised. Leave for 1h in ~500 ml de-ionised +
5ml conc HCL. Rinse with tap water then de-ionised, and put in 100% ethanol
for 1h. Replace with fresh 100% ethanol for another hour. Remove Ethanol
and air dry in covered chamber. We generally save the last ethanol wash to
re-use as the next 'first' one. I always wipe the cover-slip again with 70%
ethanol and KimTech Science 75512 tissues [wet then dry side], and leave to
dry just before use or reuse.
Haven't need to do this for a while, but it used to work. It's expensive in
time and materials though, and you ideally need some sort of rack for the
washes [easy if you have an in-house workshop].
Keith
---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.
Telephone: +44 (0)1865 287568
Email: [log in to unmask]
Web-pages: http://www.well.ox.ac.uk/cytogenetics/
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Hu Xian
Sent: 18 May 2009 07:55
To: [log in to unmask]
Subject: Protocols for reusing coverslips
Dear List,
We need to use some pretty expensive coverslips for high NA
objectives(around 8 USD per piece). We might have to reuse them :(.
Any one has any established protocol for coverslip re-use for share?
Suggestions are welcomed too.
Thanks a lot...
Regards,
Edna
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