what is your favorite method to clean cover slips?

[Zaal, Kristina (NIH/NIAMS)]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear colleagues,

I am looking for a method to clean cover slips to be used for cell culture and immunofluorescence staining. Currently, I just soak in ethanol and than flame. What is your favorite method?

Kristien J.M. Zaal, Ph.D.

Light Imaging Section

NIAMS, NIH

Bldg 50, Rm 1533

Bethesda, MD 20892

Phone [phone]; FAX [phone]

[Tamara Howard]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I prefer this protocol:

http://spectorlab.cshl.edu/acid_clean.html

Or we soak in HCl (no nitric - it kind of scares people!)
and wash as described in the Spector protocol.

Tamara

On Mon, 27 Aug 2007 11:16:03 -0400
  "Zaal, Kristina (NIH/NIAMS)" <[hidden email]> wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear colleagues,
>
>
>
> I am looking for a method to clean cover slips to be
>used for cell  culture and immunofluorescence staining.
>Currently, I just soak in  ethanol and than flame. What
>is your favorite method?
>
>
>
>
> Kristien J.M. Zaal, Ph.D.
> Light Imaging Section
> NIAMS, NIH
> Bldg 50, Rm 1533
> Bethesda, MD 20892
> Phone [phone]; FAX [phone]
>
>
>
>

***************************
Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
***************************

[mmodel]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Kristien: you don’t need coverslips to be perfectly clean for immunofluorescence, but they need to be sterile to culture cells. What you are doing sounds good enough.

 

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Zaal, Kristina (NIH/NIAMS)
Sent: Monday, August 27, 2007 11:16 AM
To: [hidden email]
Subject: what is your favorite method to clean cover slips?

 

Dear colleagues,

 

 

 

I am looking for a method to clean cover slips to be used for cell culture and immunofluorescence staining. Currently, I just soak in ethanol and than flame. What is your favorite method?

 

 

 

 

Kristien J.M. Zaal, Ph.D.

Light Imaging Section

NIAMS, NIH

Bldg 50, Rm 1533

Bethesda, MD 20892

Phone [phone]; FAX [phone]

 

 

 

[Jeffrey L. Travis]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I don't know whose method this is, but it works well:
1.)gently etch the coverslips by slowly dipping them ten times in a
solution of 10% HF / 40% nitric acid.  (use plastic vessel for this stage)
2.)follow by 30 dips in clean double distilled water (plastic vessel
here too), then
3.) 30 dips in boiling double distilled water, then
4.) 2 more  sets of 30 dips in separate beakers of distilled water;
5.) gently wick off the adherent water by touching the corner of the
coverslip to a fresh kimwipe;
6.) air dry for use or, alternatively,
7.) store the coverslips in absolute alcohol.  Flamed before use, these
will the cleanest coverslips in the known universe.

Zaal, Kristina (NIH/NIAMS) wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear colleagues,
>
>
>
> I am looking for a method to clean cover slips to be used for cell
> culture and immunofluorescence staining. Currently, I just soak in
> ethanol and than flame. What is your favorite method?
>
>
>
>
> Kristien J.M. Zaal, Ph.D.
> Light Imaging Section
> NIAMS, NIH
> Bldg 50, Rm 1533
> Bethesda, MD 20892
> Phone [phone]; FAX [phone]
>
>
>
>

[Jennifer Waters]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

 

I think cleaning coverslips is always worth it.  If nothing else you will get much better/consistent tissue culture results.  I've had users with cells that won't adhere (that normally do), look sick, etc that got better after cleaning coverslips.  I have a protocol online which I learned in the Salmon lab.

 

http://nic.med.harvard.edu/educational/index.html  (scroll down the page...)

 

Jennifer

 

On 8/27/07, Jeffrey L. Travis <[hidden email]> wrote:

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I don't know whose method this is, but it works well:
1.)gently etch the coverslips by slowly dipping them ten times in a
solution of 10% HF / 40% nitric acid.  (use plastic vessel for this stage)
2.)follow by 30 dips in clean double distilled water (plastic vessel
here too), then
3.) 30 dips in boiling double distilled water, then
4.) 2 more  sets of 30 dips in separate beakers of distilled water;
5.) gently wick off the adherent water by touching the corner of the
coverslip to a fresh kimwipe;
6.) air dry for use or, alternatively,
7.) store the coverslips in absolute alcohol.  Flamed before use, these
will the cleanest coverslips in the known universe.

Zaal, Kristina (NIH/NIAMS) wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear colleagues,
>
>
>
> I am looking for a method to clean cover slips to be used for cell
> culture and immunofluorescence staining. Currently, I just soak in
> ethanol and than flame. What is your favorite method?
>
>
>
>
> Kristien J.M. Zaal, Ph.D.
> Light Imaging Section
> NIAMS, NIH
> Bldg 50, Rm 1533
> Bethesda, MD 20892
> Phone [phone]; FAX [phone]
>
>
>
>

--
Jennifer Waters, Ph.D .
Director, Nikon Imaging Center at Harvard Medical School

[John Oreopoulos]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I find plasma cleaning for 15 minutes (if you've got a plasma cleaner) right before an experiment after a similar cleaning strategy listed on that website really helps as well. A simple test of whether or not your coverslip is super clean is to add a drop of water to it - if it's really clean, then the water droplet should spread out, whereas if it's not clean, the water will bead up on the surface.

John Oreopoulos

On 27-Aug-07, at 1:08 PM, Jennifer Waters wrote:

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

 

I think cleaning coverslips is always worth it.  If nothing else you will get much better/consistent tissue culture results.  I've had users with cells that won't adhere (that normally do), look sick, etc that got better after cleaning coverslips.  I have a protocol online which I learned in the Salmon lab.

 

http://nic.med.harvard.edu/educational/index.html  (scroll down the page...)

 

Jennifer

 

On 8/27/07, Jeffrey L. Travis <[hidden email]> wrote:

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I don't know whose method this is, but it works well:
1.)gently etch the coverslips by slowly dipping them ten times in a
solution of 10% HF / 40% nitric acid.  (use plastic vessel for this stage)
2.)follow by 30 dips in clean double distilled water (plastic vessel
here too), then
3.) 30 dips in boiling double distilled water, then
4.) 2 more  sets of 30 dips in separate beakers of distilled water;
5.) gently wick off the adherent water by touching the corner of the
coverslip to a fresh kimwipe;
6.) air dry for use or, alternatively,
7.) store the coverslips in absolute alcohol.  Flamed before use, these
will the cleanest coverslips in the known universe.

Zaal, Kristina (NIH/NIAMS) wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear colleagues,
>
>
>
> I am looking for a method to clean cover slips to be used for cell
> culture and immunofluorescence staining. Currently, I just soak in
> ethanol and than flame. What is your favorite method?
>
>
>
>
> Kristien J.M. Zaal, Ph.D.
> Light Imaging Section
> NIAMS, NIH
> Bldg 50, Rm 1533
> Bethesda, MD 20892
> Phone [phone]; FAX [phone]
>
>
>
>

--
Jennifer Waters, Ph.D .
Director, Nikon Imaging Center at Harvard Medical School

[James Pawley]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Kristien: you don't need coverslips to be perfectly clean for immunofluorescence, but they need to be sterile to culture cells. What you are doing sounds good enough.

 

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Zaal, Kristina (NIH/NIAMS)
Sent: Monday, August 27, 2007 11:16 AM
To: [hidden email]
Subject: what is your favorite method to clean cover slips?

 

Dear colleagues,

 

 

 

I am looking for a method to clean cover slips to be used for cell culture and immunofluorescence staining. Currently, I just soak in ethanol and than flame. What is your favorite method?

 

 

 

 

Kristien J.M. Zaal, Ph.D.

Light Imaging Section

NIAMS, NIH

Bldg 50, Rm 1533

Bethesda, MD 20892

Phone [phone]; FAX [phone]

 

 

 

-- 

              ****************************************
Prof. James B. Pawley,                               Ph.  608-263-3147 
Room 223, Zoology Research Building,                         FAX  [phone]
250 N. Mills St., Madison, WI, 53706  [hidden email]
"A scientist is not one who can answer questions but one who can
question answers."  Theodore Schick Jr., Skeptical Enquirer, 21-2:39

[James Pawley]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I prefer this protocol:

http://spectorlab.cshl.edu/acid_clean.html

Or we soak in HCl (no nitric - it kind of scares people!) and wash as
described in the Spector protocol.

Tamara

On Mon, 27 Aug 2007 11:16:03 -0400
  "Zaal, Kristina (NIH/NIAMS)" <[hidden email]> wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Dear colleagues,
>
>
>
>I am looking for a method to clean cover slips to be used for cell
>culture and immunofluorescence staining. Currently, I just soak in
>ethanol and than flame. What is your favorite method?
>
>
>
>
>Kristien J.M. Zaal, Ph.D.
>Light Imaging Section
>NIAMS, NIH
>Bldg 50, Rm 1533
>Bethesda, MD 20892
>Phone [phone]; FAX [phone]
>
>
>

***************************
Tamara Howard
Cell Biology & Physiology
UNM-HSC
Albuquerque, NM
***************************
--
               ****************************************
Prof. James B. Pawley,                 Ph.  608-263-3147
Room 223, Zoology Research Building,                         FAX  [phone]
250 N. Mills St., Madison, WI, 53706  [hidden email]
"A scientist is not one who can answer questions but one who can
question answers."  Theodore Schick Jr., Skeptical Enquirer, 21-2:39

[James Pawley]

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I don't know whose method this is, but it works well: 1.)gently etch
the coverslips by slowly dipping them ten times in a solution of 10%
HF / 40% nitric acid.  (use plastic vessel for this stage)
2.)follow by 30 dips in clean double distilled water (plastic vessel
here too), then
3.) 30 dips in boiling double distilled water, then
4.) 2 more  sets of 30 dips in separate beakers of distilled water;
5.) gently wick off the adherent water by touching the corner of the
coverslip to a fresh kimwipe;
6.) air dry for use or, alternatively,
7.) store the coverslips in absolute alcohol.  Flamed before use,
these will the cleanest coverslips in the known universe.

Zaal, Kristina (NIH/NIAMS) wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear
>colleagues,
>
>
>
>I am looking for a method to clean cover slips to be used for cell
>culture and immunofluorescence staining. Currently, I just soak in
>ethanol and than flame. What is your favorite method?
>
>
>
>
>Kristien J.M. Zaal, Ph.D.
>Light Imaging Section
>NIAMS, NIH
>Bldg 50, Rm 1533
>Bethesda, MD 20892
>Phone [phone]; FAX [phone]

--
               ****************************************
Prof. James B. Pawley,                 Ph.  608-263-3147
Room 223, Zoology Research Building,                         FAX  [phone]
250 N. Mills St., Madison, WI, 53706  [hidden email]
"A scientist is not one who can answer questions but one who can
question answers."  Theodore Schick Jr., Skeptical Enquirer, 21-2:39

[James Pawley]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

 

I think cleaning coverslips is always worth it.  If nothing else you will get much better/consistent tissue culture results.  I've had users with cells that won't adhere (that normally do), look sick, etc that got better after cleaning coverslips.  I have a protocol online which I learned in the Salmon lab.

 

http://nic.med.harvard.edu/educational/index.html  (scroll down the page...)

 

Jennifer

 

On 8/27/07, Jeffrey L. Travis <[hidden email]> wrote:

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I don't know whose method this is, but it works well:
1.)gently etch the coverslips by slowly dipping them ten times in a
solution of 10% HF / 40% nitric acid.  (use plastic vessel for this stage)
2.)follow by 30 dips in clean double distilled water (plastic vessel
here too), then
3.) 30 dips in boiling double distilled water, then
4.) 2 more  sets of 30 dips in separate beakers of distilled water;
5.) gently wick off the adherent water by touching the corner of the
coverslip to a fresh kimwipe;
6.) air dry for use or, alternatively,
7.) store the coverslips in absolute alcohol.  Flamed before use, these
will the cleanest coverslips in the known universe.

Zaal, Kristina (NIH/NIAMS) wrote:
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear colleagues,
>
>
>
> I am looking for a method to clean cover slips to be used for cell
> culture and immunofluorescence staining. Currently, I just soak in
> ethanol and than flame. What is your favorite method?
>
>
>
>
> Kristien J.M. Zaal, Ph.D.
> Light Imaging Section
> NIAMS, NIH
> Bldg 50, Rm 1533
> Bethesda, MD 20892
> Phone [phone]; FAX [phone]
>
>
>
>

--
Jennifer Waters, Ph.D .

Director, Nikon Imaging Center at Harvard Medical School

-- 

              ****************************************
Prof. James B. Pawley,                               Ph.  608-263-3147 
Room 223, Zoology Research Building,                         FAX  [phone]
250 N. Mills St., Madison, WI, 53706  [hidden email]
"A scientist is not one who can answer questions but one who can
question answers."  Theodore Schick Jr., Skeptical Enquirer, 21-2:39

[James Pawley]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal I find plasma cleaning for 15 minutes (if you've got a plasma cleaner) right before an experiment after a similar cleaning strategy listed on that website really helps as well. A simple test of whether or not your coverslip is super clean is to add a drop of water to it - if it's really clean, then the water droplet should spread out, whereas if it's not clean, the water will bead up on the surface.

John Oreopoulos

On 27-Aug-07, at 1:08 PM, Jennifer Waters wrote:

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

 

I think cleaning coverslips is always worth it.  If nothing else you will get much better/consistent tissue culture results.  I've had users with cells that won't adhere (that normally do), look sick, etc that got better after cleaning coverslips.  I have a protocol online which I learned in the Salmon lab.

 

http://nic.med.harvard.edu/educational/index.html  (scroll down the page...)

 

Jennifer

 

On 8/27/07, Jeffrey L. Travis <[hidden email]> wrote:

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

I don't know whose method this is, but it works well:
1.)gently etch the coverslips by slowly dipping them ten times in a
solution of 10% HF / 40% nitric acid.  (use plastic vessel for this stage)
2.)follow by 30 dips in clean double distilled water (plastic vessel
here too), then
3.) 30 dips in boiling double distilled water, then
4.) 2 more  sets of 30 dips in separate beakers of distilled water;
5.) gently wick off the adherent water by touching the corner of the
coverslip to a fresh kimwipe;
6.) air dry for use or, alternatively,
7.) store the coverslips in absolute alcohol.  Flamed before use, these
will the cleanest coverslips in the known universe.

Zaal, Kristina (NIH/NIAMS) wrote:
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear colleagues,
>
>
>
> I am looking for a method to clean cover slips to be used for cell
> culture and immunofluorescence staining. Currently, I just soak in
> ethanol and than flame. What is your favorite method?
>
>
>
>
> Kristien J.M. Zaal, Ph.D.
> Light Imaging Section
> NIAMS, NIH
> Bldg 50, Rm 1533
> Bethesda, MD 20892
> Phone [phone]; FAX [phone]
>
>
>
>

--
Jennifer Waters, Ph.D .
Director, Nikon Imaging Center at Harvard Medical School

-- 

              ****************************************
Prof. James B. Pawley,                               Ph.  608-263-3147 
Room 223, Zoology Research Building,                         FAX  [phone]
250 N. Mills St., Madison, WI, 53706  [hidden email]
"A scientist is not one who can answer questions but one who can
question answers."  Theodore Schick Jr., Skeptical Enquirer, 21-2:39