Clean slides and cover slips

[Zucker.Robert]

I was using some good quality pre-cleaned microscope slides and noticed
that they  are not clean but very "dirty".  Good cover slips ( 1.5 )
also have some dirt on them.
Does anyone have a protocol for cleaning  precleaned slides and
coverslips and a source to perhaps buy good slides/coverlsips. .
It is very hard to look at small particles in a field with dirt and
streaks on the so called pre-cleaned slides
thanks
bob



Robert M. Zucker, PhD
U.S. Environmental Protection Agency
Office of Research and Development
National Health and Environmental Effects Research Laboratory.
Reproductive Toxicology Division
Telephone: [phone]   Fax: [phone]
e-mail: [hidden email]

Mail address: USEPA,ORD,NHEERL,RTD
Developmental Biology Branch ( MD 67)
Research Triangle Park, North Carolina, 27711

Shipping address:
2525 E.NC Highway 54
Durham, NC, 27713

[Keith Morris]

Hi Robert,

Talked about this recently on the listserver, so have an instant reply:

[This wash sequence is used for new 'pre-washed' glass slides from boxes]:
Soak the slides overnight in 5ml Teepol/RenClean in ~500 ml MillQ water [you
need a large 100 slide slotted rack to hold the slides apart and upright in
the tank, and to lift them in/out]. Next morning wash off detergent gently
with de-ionised tap water* then MillQ**. Leave for 1h in ~500 ml MillQ + 5ml
conc HCL. Rinse with de-ionised tap water then MillQ, 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 don't pre-wash cover-slips, but prior to use I always wipe the cover-slip
with 70% ethanol and KimTech Science 75512 tissues*** [70% alcohol wet then
dry side], and leave cover-slip to dry resting against a rack just before
use. The slides are stored in their dry racks/covered tank and removed as
needed.

You need some sort of 100 slide rack and tank [with a lid] for the washes
[ours have been around since the year dot].We use VWR [formally BDH]
'washed,cleaned, polished Superfrost' [cat no. 631-0102, the labeling area
is available in 7 colours including pink 631-0104]. Our borosilicate
cover-slips are also VWR [e.g. catalogue 631-0138].

Tip: If you have to dry a wet [PBS/SSC/water] slide after processing, use a
hand puffer**** to blow the liquid off the slide, then airdry upright in a
slotted slide holder [if complete drying is required] and apply
cover-slip+mountant. You can wipe the base & edges of the slide with a
Kimtech tissue, avoiding the specimen area. This helps you to eliminate
'water marks' on the slide surface. Avoid compressed air aerosol cans though
as these are expensive to use and can fire propellent onto the slide
surface, making a worse mess than the drying water/pbs.

Keith

*could be tap water
**could be lower grade 'deionised' water
We just happen to have de-ionised supplied to a tap in the lab and a local
MillQ water purifier. The times are 'minimum's, so you can leave the slides
in alcohol/acid etc.. for longer if you are busy in the lab.

***These expensive tissues are ideal, Blue-rolls are too inflexible/abrasive
and softer tissues give off fluff.

****or better still low pressure compressed air [air outlet via clear
plastic tubing and a in-line disposable submicron filter, plus wear safety
glasses] to blow the liquid off the slide, then airdry upright in a slotted
slide holder [if complete drying is required] and apply cover-slip+mountant.
Sadly our building has no gas or compressed air taps in the lab owing to
'health & safety' issues, so I use a very large photographic hand puffer
these days [no brush] - it's a natty number shaped like a space rocket, and
stands up on its fins:

http://www.amazon.co.uk/Giottos-GTAA-1901-Large-Rocket/dp/B000RGM762
http://www.amazon.co.uk/Giottos-GTAA1900-Rocket-Air-Blower/dp/B00017LSPI/ref
=pd_cp_ce_0



---------------------------------------------------------------------------
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:  [phone]
Email:  [hidden email]
Web-pages: http://www.well.ox.ac.uk/cytogenetics/



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of [hidden email]
Sent: 25 June 2009 01:44
To: [hidden email]
Subject: Clean slides and cover slips

I was using some good quality pre-cleaned microscope slides and noticed
that they  are not clean but very "dirty".  Good cover slips ( 1.5 )
also have some dirt on them.
Does anyone have a protocol for cleaning  precleaned slides and
coverslips and a source to perhaps buy good slides/coverlsips. .
It is very hard to look at small particles in a field with dirt and
streaks on the so called pre-cleaned slides
thanks
bob



Robert M. Zucker, PhD
U.S. Environmental Protection Agency
Office of Research and Development
National Health and Environmental Effects Research Laboratory.
Reproductive Toxicology Division
Telephone: [phone]   Fax: [phone]
e-mail: [hidden email]

Mail address: USEPA,ORD,NHEERL,RTD
Developmental Biology Branch ( MD 67)
Research Triangle Park, North Carolina, 27711

Shipping address:
2525 E.NC Highway 54
Durham, NC, 27713

[Goodhouse, Joseph G.]

 


Joe Goodhouse
Confocal Core Lab Manager
Dept. of Molecular Biology
Princeton University
609-258-5432

Visit us at http://www.molbio1.princeton.edu/facility/confocal/   


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of [hidden email]
Sent: Wednesday, June 24, 2009 8:44 PM
To: [hidden email]
Subject: Clean slides and cover slips

I was using some good quality pre-cleaned microscope slides and noticed
that they  are not clean but very "dirty".  Good cover slips ( 1.5 )
also have some dirt on them.
Does anyone have a protocol for cleaning  precleaned slides and
coverslips and a source to perhaps buy good slides/coverlsips. .
It is very hard to look at small particles in a field with dirt and
streaks on the so called pre-cleaned slides thanks bob



Robert M. Zucker, PhD
U.S. Environmental Protection Agency
Office of Research and Development
National Health and Environmental Effects Research Laboratory.
Reproductive Toxicology Division
Telephone: [phone]   Fax: [phone]
e-mail: [hidden email]

Mail address: USEPA,ORD,NHEERL,RTD
Developmental Biology Branch ( MD 67)
Research Triangle Park, North Carolina, 27711

Shipping address:
2525 E.NC Highway 54
Durham, NC, 27713

[Goodhouse, Joseph G.]

Bob,
        I use acid alcohol for this.  95% ETOH with 5% 1N HCL. I soak
for several hours or overnight.  Rinse with ETOH Then DI water.  All
slides appear to have an oil residue on them.


Joe Goodhouse
Confocal Core Lab Manager
Dept. of Molecular Biology
Princeton University
609-258-5432

Visit us at http://www.molbio1.princeton.edu/facility/confocal/   


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of [hidden email]
Sent: Wednesday, June 24, 2009 8:44 PM
To: [hidden email]
Subject: Clean slides and cover slips

I was using some good quality pre-cleaned microscope slides and noticed
that they  are not clean but very "dirty".  Good cover slips ( 1.5 )
also have some dirt on them.
Does anyone have a protocol for cleaning  precleaned slides and
coverslips and a source to perhaps buy good slides/coverlsips. .
It is very hard to look at small particles in a field with dirt and
streaks on the so called pre-cleaned slides thanks bob



Robert M. Zucker, PhD
U.S. Environmental Protection Agency
Office of Research and Development
National Health and Environmental Effects Research Laboratory.
Reproductive Toxicology Division
Telephone: [phone]   Fax: [phone]
e-mail: [hidden email]

Mail address: USEPA,ORD,NHEERL,RTD
Developmental Biology Branch ( MD 67)
Research Triangle Park, North Carolina, 27711

Shipping address:
2525 E.NC Highway 54
Durham, NC, 27713

[vb-2]

Hi Robert,

Yes, most of coverglass is quite dirty (most of standard #1.5 coverslips
from many suppliers, even the German one from the Hecht_Assistent;  also
very dirty BK-7 coverglass, Nuncbrand LabTek I or II, Grace Biolabs 16-well
chambered coverglass). The MatTek glass bottom dishes are cleaner but in
many instances the coverglass is "angled", e.g. mounted not strictly
horizontal giving you out-of-focus corners when you image anything that
mounted on the surface.

I attached one rare paper from China that is surprisingly very remarkable,
in technical terms.  ... as NIH would never fund such a project (but USDOE
might have contributed to  the study).

It is a shame that postdocs (and/or skilled technicians) have to kill time
cleaning coverglass... Maybe NIH can provide funds for a "Coverglass
Cleaning Biotech Startup Company"

Good luck,

Vitaly

NCI-Frederick
301-846-6575

PS As my original posting was rejected 'cos I attached pdf file, I will send
it to upon request

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of [hidden email]
Sent: Wednesday, June 24, 2009 7:44 PM
To: [hidden email]
Subject: Clean slides and cover slips

I was using some good quality pre-cleaned microscope slides and noticed
that they  are not clean but very "dirty".  Good cover slips ( 1.5 )
also have some dirt on them.
Does anyone have a protocol for cleaning  precleaned slides and
coverslips and a source to perhaps buy good slides/coverlsips. .
It is very hard to look at small particles in a field with dirt and
streaks on the so called pre-cleaned slides
thanks
bob



Robert M. Zucker, PhD
U.S. Environmental Protection Agency
Office of Research and Development
National Health and Environmental Effects Research Laboratory.
Reproductive Toxicology Division
Telephone: [phone]   Fax: [phone]
e-mail: [hidden email]

Mail address: USEPA,ORD,NHEERL,RTD
Developmental Biology Branch ( MD 67)
Research Triangle Park, North Carolina, 27711

Shipping address:
2525 E.NC Highway 54
Durham, NC, 27713

[Rietdorf, Jens]

Dear bob,

Some time ago, we have been ordering pre-cleaned cover glasses through
http://www.primeglass.de/  (no comm. Interest); some neuronal cell lines
refuse to grow on non pre-cleaned glasses and they certify their glasses
for growth of delicate cell lines (and it worked). No idea how
'optically' clean those glasses are, but their cleaning procedure
involves some acid and alcohol treatments like mentioned in some of the
replies to your initial posting. May be worth a try.


cheers, jens

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of [hidden email]
Sent: Thursday, June 25, 2009 2:44 AM
To: [hidden email]
Subject: Clean slides and cover slips

I was using some good quality pre-cleaned microscope slides and noticed
that they  are not clean but very "dirty".  Good cover slips ( 1.5 )
also have some dirt on them.
Does anyone have a protocol for cleaning  precleaned slides and
coverslips and a source to perhaps buy good slides/coverlsips. .
It is very hard to look at small particles in a field with dirt and
streaks on the so called pre-cleaned slides
thanks
bob



Robert M. Zucker, PhD
U.S. Environmental Protection Agency
Office of Research and Development
National Health and Environmental Effects Research Laboratory.
Reproductive Toxicology Division
Telephone: [phone]   Fax: [phone]
e-mail: [hidden email]

Mail address: USEPA,ORD,NHEERL,RTD
Developmental Biology Branch ( MD 67)
Research Triangle Park, North Carolina, 27711

Shipping address:
2525 E.NC Highway 54
Durham, NC, 27713