coverslips for cell culture

[Fredrik Wermeling]

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

Hi!

 

I’m planning to grow adherent cells for later staining with fluorescently labelled antibodies. Previously I’ve done this by adding a sterile round coverslip in a 12 well plate and kept the cells on the coverslip through out the staining and fixation and then just "mounted" them on a microscope slide. My questions are:

- Do you have any suggestions for which coverslips to purchase, I guess they have to be thin enough and without auto fluorescence.

- Is there any alternative? I saw that Nunc sells a Lab tek product that sounds like what I´m looking for. 

 

Thanks a lot

/ Fredrik

 

[Sandrine Pouvreau]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Hi.
I'm doing this kind of things with Hek cells or myotubes on cell culture round dishes with glass coverslip bottom of several kind. The coverslip has to be 0.17mm thin, but I never had any autofluorescence problem. I'm for instance using (from Fisher Scientist) Delta TPG dish, 0.17mm. They have a thick ring of plastic surrounding the glass coverslip, so it is hard to focus on the cells that are on the edge, but I just cut it with scissor and it works pretty well.
I wish you good luck
Sandrine

Fredrik Wermeling <[hidden email]>
Sent by: Confocal Microscopy List <[hidden email]>

10/25/2007 03:13 PM

Please respond to
Confocal Microscopy List <[hidden email]>

To	[hidden email]
cc
Subject	coverslips for cell culture

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Hi!
 
I’m planning to grow adherent cells for later staining with fluorescently labelled antibodies. Previously I’ve done this by adding a sterile round coverslip in a 12 well plate and kept the cells on the coverslip through out the staining and fixation and then just "mounted" them on a microscope slide. My questions are:
- Do you have any suggestions for which coverslips to purchase, I guess they have to be thin enough and without auto fluorescence.
- Is there any alternative? I saw that Nunc sells a Lab tek product that sounds like what I´m looking for.
 
Thanks a lot
/ Fredrik

 

[Kurt Thorn]

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

You want to use coverslips that are 0.17 mm thick - number 1.5
coverslips are the closest in thickness.

You can also use coverslip bottom plates or coverlsip bottom dishes -
these are very convenient if you want to image the cells live but may be
more of a pain for fixing and staining.  Glass-bottom-dishes.com sell a
number of different varieties.

Kurt

Fredrik Wermeling wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi!
>
>  
>
> I’m planning to grow adherent cells for later staining with
> fluorescently labelled antibodies. Previously I’ve done this by adding
> a sterile round coverslip in a 12 well plate and kept the cells on the
> coverslip through out the staining and fixation and then just
> "mounted" them on a microscope slide. My questions are:
>
> - Do you have any suggestions for which coverslips to purchase, I
> guess they have to be thin enough and without auto fluorescence.
>
> - Is there any alternative? I saw that Nunc sells a Lab tek product
> that sounds like what I´m looking for.
>
>  
>
> Thanks a lot
>
> / Fredrik
>
>  
>

--
Kurt Thorn, PhD
Director, Nikon Imaging Center
University of California San Francisco

UCSF MC 2140
Genentech Hall Room S252
600 16th St.
San Francisco, CA 94158-2517

http://nic.ucsf.edu
phone [phone]
fax   [phone]

[Stephen Cody]

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

>You want to use coverslips that are 0.17 mm thick - number 1.5
>coverslips are the closest in thickness.

The trouble with No. 1.5 is that there is great variation in thickness.
The industry standard for No. 1.50 is:
#1.5 = 0.160  -  0.190

Please review the achieves under Subject: "Coverslip - Good News'

This is quite a large range, and 0.190 mm thick coverslips will cause detrimental spherical aberration.
 
Both Menzel and Assistent (Hecht Glaswarenfabrik [Thanks Beat]) sell a better standard thickness for confocal microscopy it is called:
"170 +/- 0.01mm"
 
Many of the distributors may not know about this sorting of coverslip, but it does exist. You may need to convince them to ask their suppliers about it.
 
If you still have trouble you can order them from Lomb in Australia, they will deliver.

I order "0.170 +/- 0.01mm" coverslips for confocal through
Lomb Scientific www.lomb.com.au . This is an Australian distributor
of Menzel Glass. Lomb will also cut coverslips to special custom sizes
(larger bulk orders are more economical).

You can contact Peter directly on [hidden email]  for product
information, order codes etc..

Or as Peter suggests he is not always available [hidden email]  may
work better sometimes.

I have no commercial affiliation with any of the companies listed. I can prove it, have a look at the car I drive! :)
 
Cheers
Stephen H. Cody
Microscopy Manager
Central Resource for Advanced Microscopy
Ludwig Institute for Cancer Research
PO Box 2008 Royal Melbourne Hospital
Parkville, Victoria,      3050
Australia
Tel: [phone]    Fax: [phone]
email: [hidden email]
www.ludwig.edu.au/labs/confocal.html
www.ludwig.edu.au/confocal

Tip: Learn how to receive reminders about you microscope booking:
http://www.ludwig.edu.au/confocal/Local/Booking_Hint.htm Type your signature here

________________________________

From: Confocal Microscopy List on behalf of Kurt Thorn
Sent: Fri 26/10/2007 6:33 AM
To: [hidden email]
Subject: Re: coverslips for cell culture



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

You want to use coverslips that are 0.17 mm thick - number 1.5
coverslips are the closest in thickness.

You can also use coverslip bottom plates or coverlsip bottom dishes -
these are very convenient if you want to image the cells live but may be
more of a pain for fixing and staining.  Glass-bottom-dishes.com sell a
number of different varieties.

Kurt

Fredrik Wermeling wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi!
>
>
>
> I'm planning to grow adherent cells for later staining with
> fluorescently labelled antibodies. Previously I've done this by adding
> a sterile round coverslip in a 12 well plate and kept the cells on the
> coverslip through out the staining and fixation and then just
> "mounted" them on a microscope slide. My questions are:
>
> - Do you have any suggestions for which coverslips to purchase, I
> guess they have to be thin enough and without auto fluorescence.
>
> - Is there any alternative? I saw that Nunc sells a Lab tek product
> that sounds like what I´m looking for.
>
>
>
> Thanks a lot
>
> / Fredrik
>
>
>

--
Kurt Thorn, PhD
Director, Nikon Imaging Center
University of California San Francisco

UCSF MC 2140
Genentech Hall Room S252
600 16th St.
San Francisco, CA 94158-2517

http://nic.ucsf.edu <http://nic.ucsf.edu/>
phone [phone]
fax   [phone]

[Donnelly, Tom]

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

The  Titertek II  ( Nunc) 4 and 8 well dishes have # 1.5 coverslip bottoms and they work well.

 

I would suggest looking at www.willcowells.com  they make very high quality coverslip bottom 35mm dishes with both 12mm and 22mm 170um coverslips. They also make a "kit" which would allow you to grow cells on the coverslip then insert it into the machined 35mm dish.

 

No commercial interest in either.

 

Tom

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Fredrik Wermeling
Sent: Thursday, October 25, 2007 1:13 PM
To: [hidden email]
Subject: coverslips for cell culture

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

Hi!

 

I’m planning to grow adherent cells for later staining with fluorescently labelled antibodies. Previously I’ve done this by adding a sterile round coverslip in a 12 well plate and kept the cells on the coverslip through out the staining and fixation and then just "mounted" them on a microscope slide. My questions are:

- Do you have any suggestions for which coverslips to purchase, I guess they have to be thin enough and without auto fluorescence.

- Is there any alternative? I saw that Nunc sells a Lab tek product that sounds like what I´m looking for. 

 

Thanks a lot

/ Fredrik

 

[Kurt Thorn]

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

Does anyone know of a US source for these?  When I this came up earlier
on the list I did some searching for a US distributor but came up empty
handed.  I agree that if you can get these it would be a big improvement
on using #1.5 coverslips.

Kurt
> Both Menzel and Assistent (Hecht Glaswarenfabrik [Thanks Beat]) sell a better standard thickness for confocal microscopy it is called:
> "170 +/- 0.01mm"
>  

--
Kurt Thorn, PhD
Director, Nikon Imaging Center
University of California San Francisco

UCSF MC 2140
Genentech Hall Room S252
600 16th St.
San Francisco, CA 94158-2517

http://nic.ucsf.edu
phone [phone]
fax   [phone]

[Stephen Cody]

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

Dear Kurt,
 
I suspect Menzel and Assistent  (Hecht) are the only manufacturers of coverslip glass. I suspect other companies buy the glass, perhaps cut it, and rebrand it.  This is certainly what Lomb in Australia do, they are quite open about it. If there are other manufacturers of coverslip glass I'd be happy to be corrected. So if you buy from a particular company or supplier, I suspect is is probably Menzel or Assistent glass anyway, and you need to put a little pressure on your supplier to contact their manufacturer and order the correct thickness.
 
Cheers
Stephen H. Cody
Microscopy Manager
Central Resource for Advanced Microscopy
Ludwig Institute for Cancer Research
PO Box 2008 Royal Melbourne Hospital
Parkville, Victoria,      3050
Australia
Tel: [phone]    Fax: [phone]
email: [hidden email]
www.ludwig.edu.au/labs/confocal.html
www.ludwig.edu.au/confocal

Tip: Learn how to receive reminders about you microscope booking:
http://www.ludwig.edu.au/confocal/Local/Booking_Hint.htm Type your signature here

________________________________

From: Confocal Microscopy List on behalf of Kurt Thorn
Sent: Fri 26/10/2007 9:42 AM
To: [hidden email]
Subject: Re: coverslips for cell culture



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

Does anyone know of a US source for these?  When I this came up earlier
on the list I did some searching for a US distributor but came up empty
handed.  I agree that if you can get these it would be a big improvement
on using #1.5 coverslips.

Kurt
> Both Menzel and Assistent (Hecht Glaswarenfabrik [Thanks Beat]) sell a better standard thickness for confocal microscopy it is called:
> "170 +/- 0.01mm"
>

--
Kurt Thorn, PhD
Director, Nikon Imaging Center
University of California San Francisco

UCSF MC 2140
Genentech Hall Room S252
600 16th St.
San Francisco, CA 94158-2517

http://nic.ucsf.edu <http://nic.ucsf.edu/>
phone [phone]
fax   [phone]

[Glen MacDonald-2]

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

Erie Glass, which sells under their own label as well as sells to other companies, actually owns Menzel.  
The glass sold by both is now made by Erie Electroverre in Romont, Switzerland.   Erie has other plants
around the world and is now part of Thermo-Fisher.

How long before there are only 2 corporations left in the world?

Regards,
Glen

Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923
[phone]

On Fri, 26 Oct 2007, Stephen Cody wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear Kurt,
>
> I suspect Menzel and Assistent  (Hecht) are the only manufacturers of coverslip glass. I suspect other
companies buy the glass, perhaps cut it, and rebrand it.  This is certainly what Lomb in Australia do,
they are quite open about it. If there are other manufacturers of coverslip glass I'd be happy to be
corrected. So if you buy from a particular company or supplier, I suspect is is probably Menzel or
Assistent glass anyway, and you need to put a little pressure on your supplier to contact their
manufacturer and order the correct thickness.

>
> Cheers
> Stephen H. Cody
> Microscopy Manager
> Central Resource for Advanced Microscopy
> Ludwig Institute for Cancer Research
> PO Box 2008 Royal Melbourne Hospital
> Parkville, Victoria,      3050
> Australia
> Tel: [phone]    Fax: [phone]
> email: [hidden email]
> www.ludwig.edu.au/labs/confocal.html
> www.ludwig.edu.au/confocal
>
> Tip: Learn how to receive reminders about you microscope booking:
> http://www.ludwig.edu.au/confocal/Local/Booking_Hint.htm Type your signature here
>
> ________________________________
>
> From: Confocal Microscopy List on behalf of Kurt Thorn
> Sent: Fri 26/10/2007 9:42 AM
> To: [hidden email]
> Subject: Re: coverslips for cell culture
>
>
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Does anyone know of a US source for these?  When I this came up earlier
> on the list I did some searching for a US distributor but came up empty
> handed.  I agree that if you can get these it would be a big improvement
> on using #1.5 coverslips.
>
> Kurt
>> Both Menzel and Assistent (Hecht Glaswarenfabrik [Thanks Beat]) sell a better standard thickness

for confocal microscopy it is called:

>> "170 +/- 0.01mm"
>>
>
> --
> Kurt Thorn, PhD
> Director, Nikon Imaging Center
> University of California San Francisco
>
> UCSF MC 2140
> Genentech Hall Room S252
> 600 16th St.
> San Francisco, CA 94158-2517
>
> http://nic.ucsf.edu <http://nic.ucsf.edu/>
> phone [phone]
> fax   [phone]
>

[Sarah Aubut]

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

I have used the Nunc chambered coverglass, and they have worked
excellent for me with viewing living cells and labeling them while doing
time lapse. These are like the chamber slide, except the bottom is the
coverglass and the chambers cannot be removed, which makes them perfect
for viewing live cells. Make sure you get the LabTek II, not the Labtek
I, as the II's have the 1.5 thickness. I order mine directly from
Nalgene-Nunc without a problem. The only issue I had was they are
shorter then a normal slide, so did not fit into my stage, but as I use
them all the time, I invested in a Prior stage that is designed for
them. I'm sure you can easily rig your normal stage if they don't fit.

Sarah Aubut
Lab Technician, Microscopy Suite
Canadian Forestry Service
Sault Ste Marie, ON
Canada

[hidden email]

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Glen H MacDonald
Sent: Friday, October 26, 2007 12:16 AM
To: [hidden email]
Subject: Re: coverslips for cell culture

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

Erie Glass, which sells under their own label as well as sells to other
companies, actually owns Menzel.  
The glass sold by both is now made by Erie Electroverre in Romont,
Switzerland.   Erie has other plants
around the world and is now part of Thermo-Fisher.

How long before there are only 2 corporations left in the world?

Regards,
Glen

Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center Box 357923 University
of Washington Seattle, WA 98195-7923
[phone]

On Fri, 26 Oct 2007, Stephen Cody wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear Kurt,
>
> I suspect Menzel and Assistent  (Hecht) are the only manufacturers of
> coverslip glass. I suspect other
companies buy the glass, perhaps cut it, and rebrand it.  This is
certainly what Lomb in Australia do, they are quite open about it. If
there are other manufacturers of coverslip glass I'd be happy to be
corrected. So if you buy from a particular company or supplier, I
suspect is is probably Menzel or Assistent glass anyway, and you need to
put a little pressure on your supplier to contact their manufacturer and
order the correct thickness.

>
> Cheers
> Stephen H. Cody
> Microscopy Manager
> Central Resource for Advanced Microscopy Ludwig Institute for Cancer
> Research PO Box 2008 Royal Melbourne Hospital
> Parkville, Victoria,      3050
> Australia
> Tel: [phone]    Fax: [phone]
> email: [hidden email]
> www.ludwig.edu.au/labs/confocal.html
> www.ludwig.edu.au/confocal
>
> Tip: Learn how to receive reminders about you microscope booking:
> http://www.ludwig.edu.au/confocal/Local/Booking_Hint.htm Type your
> signature here
>
> ________________________________
>
> From: Confocal Microscopy List on behalf of Kurt Thorn
> Sent: Fri 26/10/2007 9:42 AM
> To: [hidden email]
> Subject: Re: coverslips for cell culture
>
>
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Does anyone know of a US source for these?  When I this came up
> earlier on the list I did some searching for a US distributor but came

> up empty handed.  I agree that if you can get these it would be a big
> improvement on using #1.5 coverslips.
>
> Kurt
>> Both Menzel and Assistent (Hecht Glaswarenfabrik [Thanks Beat]) sell
>> a better standard thickness
for confocal microscopy it is called:

>> "170 +/- 0.01mm"
>>
>
> --
> Kurt Thorn, PhD
> Director, Nikon Imaging Center
> University of California San Francisco
>
> UCSF MC 2140
> Genentech Hall Room S252
> 600 16th St.
> San Francisco, CA 94158-2517
>
> http://nic.ucsf.edu <http://nic.ucsf.edu/> phone [phone]
> fax   [phone]
>

[rjpalmer]

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

Couldn't resist an off-list post on this one. Before retirement (ca
15 years), I expect to see all of US commerce to be roughly equally
divided between Invitrogen and Halliburton. :) :)  The rest of the
world should be sorted out within another 15 - that is, if anything
remains to be sorted....(not talking about commerce there...)

Have you seen the film "Idiocracy"?


>
>How long before there are only 2 corporations left in the world?
>
>Regards,
>Glen

--
Robert J. Palmer Jr., Ph.D.
Natl Inst Dental Craniofacial Res - Natl Insts Health
Oral Infection and Immunity Branch
Bldg 30, Room 310
30 Convent Drive
Bethesda MD 20892
ph 301-594-0025
fax [phone]

[rjpalmer]

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

So much for off-list  :) :)  Please don't turn me in to my employer (GW)...

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Couldn't resist an off-list post on this one. Before retirement (ca
>15 years), I expect to see all of US commerce to be roughly equally
>divided between Invitrogen and Halliburton. :) :)  The rest of the
>world should be sorted out within another 15 - that is, if anything
>remains to be sorted....(not talking about commerce there...)
>
>Have you seen the film "Idiocracy"?
>
>>
>>How long before there are only 2 corporations left in the world?
>>
>>Regards,
>>Glen
>
>--
>Robert J. Palmer Jr., Ph.D.
>Natl Inst Dental Craniofacial Res - Natl Insts Health
>Oral Infection and Immunity Branch
>Bldg 30, Room 310
>30 Convent Drive
>Bethesda MD 20892
>ph 301-594-0025
>fax [phone]

--
Robert J. Palmer Jr., Ph.D.
Natl Inst Dental Craniofacial Res - Natl Insts Health
Oral Infection and Immunity Branch
Bldg 30, Room 310
30 Convent Drive
Bethesda MD 20892
ph 301-594-0025
fax [phone]

[Glen MacDonald-2]

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

the Nunc II chambers are useful, but the thickness still varies.  I  
bought a blank insert for the our Prior stage and had it machined to  
take 60 mm coverslips in x dimension and Nunc chambers in the y  
direction.  A small trim of the corners at the intersection of the 2  
rectangular openings allows a 35 mm glass bottom dish to be used, as  
well.  a 1.5 cm semi-circle enlargement at the ends of the  
rectangular openings allow access to the lens to apply immersion media.

Glen

On Oct 26, 2007, at 4:53 AM, Aubut, Sarah wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> I have used the Nunc chambered coverglass, and they have worked
> excellent for me with viewing living cells and labeling them while  
> doing
> time lapse. These are like the chamber slide, except the bottom is the
> coverglass and the chambers cannot be removed, which makes them  
> perfect
> for viewing live cells. Make sure you get the LabTek II, not the  
> Labtek
> I, as the II's have the 1.5 thickness. I order mine directly from
> Nalgene-Nunc without a problem. The only issue I had was they are
> shorter then a normal slide, so did not fit into my stage, but as I  
> use
> them all the time, I invested in a Prior stage that is designed for
> them. I'm sure you can easily rig your normal stage if they don't fit.
>
> Sarah Aubut
> Lab Technician, Microscopy Suite
> Canadian Forestry Service
> Sault Ste Marie, ON
> Canada
>
> [hidden email]
>
> -----Original Message-----
> From: Confocal Microscopy List  
> [mailto:[hidden email]] On
> Behalf Of Glen H MacDonald
> Sent: Friday, October 26, 2007 12:16 AM
> To: [hidden email]
> Subject: Re: coverslips for cell culture
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Erie Glass, which sells under their own label as well as sells to  
> other
> companies, actually owns Menzel.
> The glass sold by both is now made by Erie Electroverre in Romont,
> Switzerland.   Erie has other plants
> around the world and is now part of Thermo-Fisher.
>
> How long before there are only 2 corporations left in the world?
>
> Regards,
> Glen
>
> Glen MacDonald
> Core for Communication Research
> Virginia Merrill Bloedel Hearing Research Center Box 357923 University
> of Washington Seattle, WA 98195-7923
> [phone]
>
> On Fri, 26 Oct 2007, Stephen Cody wrote:
>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Dear Kurt,
>>
>> I suspect Menzel and Assistent  (Hecht) are the only manufacturers of
>> coverslip glass. I suspect other
> companies buy the glass, perhaps cut it, and rebrand it.  This is
> certainly what Lomb in Australia do, they are quite open about it. If
> there are other manufacturers of coverslip glass I'd be happy to be
> corrected. So if you buy from a particular company or supplier, I
> suspect is is probably Menzel or Assistent glass anyway, and you  
> need to
> put a little pressure on your supplier to contact their  
> manufacturer and
> order the correct thickness.
>>
>> Cheers
>> Stephen H. Cody
>> Microscopy Manager
>> Central Resource for Advanced Microscopy Ludwig Institute for Cancer
>> Research PO Box 2008 Royal Melbourne Hospital
>> Parkville, Victoria,      3050
>> Australia
>> Tel: [phone]    Fax: [phone]
>> email: [hidden email]
>> www.ludwig.edu.au/labs/confocal.html
>> www.ludwig.edu.au/confocal
>>
>> Tip: Learn how to receive reminders about you microscope booking:
>> http://www.ludwig.edu.au/confocal/Local/Booking_Hint.htm Type your
>> signature here
>>
>> ________________________________
>>
>> From: Confocal Microscopy List on behalf of Kurt Thorn
>> Sent: Fri 26/10/2007 9:42 AM
>> To: [hidden email]
>> Subject: Re: coverslips for cell culture
>>
>>
>>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Does anyone know of a US source for these?  When I this came up
>> earlier on the list I did some searching for a US distributor but  
>> came
>
>> up empty handed.  I agree that if you can get these it would be a big
>> improvement on using #1.5 coverslips.
>>
>> Kurt
>>> Both Menzel and Assistent (Hecht Glaswarenfabrik [Thanks Beat]) sell
>>> a better standard thickness
> for confocal microscopy it is called:
>>> "170 +/- 0.01mm"
>>>
>>
>> --
>> Kurt Thorn, PhD
>> Director, Nikon Imaging Center
>> University of California San Francisco
>>
>> UCSF MC 2140
>> Genentech Hall Room S252
>> 600 16th St.
>> San Francisco, CA 94158-2517
>>
>> http://nic.ucsf.edu <http://nic.ucsf.edu/> phone [phone]
>> fax   [phone]
>>

[Claire Brown]

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

We use 35 mm glass bottom dishes from World Precision Instruments. They say
on their web page they are 0.17 mm +- 0.01 mm. They are also much cheaper
than other suppliers if you buy them in boxes of 100.

No commercial interest.

Claire

[Michael Weber-4]

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

fyi (no commercial interest):

http://www.zeiss.de/C1256A770030BCE0/WebViewAllE/162CCB55A50A433FC12573A8002B17D9

The press release contains quite some side-kicks to Leica. And the
technique sounds kind of STORM-like. Does somebody has further information?

cheers,
Michael

[Martin Wessendorf]

Search the CONFOCAL archive at
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Michael Weber wrote:

> http://www.zeiss.de/C1256A770030BCE0/WebViewAllE/162CCB55A50A433FC12573A8002B17D9 

> The press release contains quite some side-kicks to Leica. And the
> technique sounds kind of STORM-like. Does somebody has further information?

I think you're right.  Looks as if they've teamed up with the group
headed by Eric Betzig.  He had the paper in Science last year:

"Imaging Intracellular Fluorescent Proteins at Nanometer Resolution"
Eric Betzig, George H. Patterson, Rachid Sougrat, O. Wolf Lindwasser,
Scott Olenych, Juan S. Bonifacino, Michael W. Davidson, Jennifer
Lippincott-Schwartz, and Harald F. Hess
Science 15 September 2006 313: 1642-1645

If that's it, it seems like a powerful technique, lower-cost than the
Leica system, but unfortunately also very slow.  Also, it appears only
applicable to photoactivatable fluorescent proteins (or dyes) that will
change their spectrum to a longer wavelength when excited with a shorter
wavelength.  It has the advantage of using much less intense
illumination than the Leica system but I don't know of anyone using it
for live-cell work.

Martin
--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455             E-mail: martinw[at]med.umn.edu

[John Oreopoulos]

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal STORM and PALM are similar. Here is the original publication on the PALM technique:

Betzig, E., G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess. 2006. Imaging intracellular fluorescent proteins at nanometer resolution. Science 313:1642-1645. 

John Oreopoulos, BSc,

PhD Candidate

University of Toronto

Institute For Biomaterials and Biomedical Engineering

Centre For Studies in Molecular Imaging

Tel: W:416-946-5022

On 5-Dec-07, at 11:04 AM, Michael Weber wrote:

Search the CONFOCAL archive at

http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

fyi (no commercial interest):

http://www.zeiss.de/C1256A770030BCE0/WebViewAllE/162CCB55A50A433FC12573A8002B17D9

The press release contains quite some side-kicks to Leica. And the technique sounds kind of STORM-like. Does somebody has further information?

cheers,

Michael

[Allan Kachelmeier]

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

It looks like the speed of the Palm/Storm/Palmira technique is being tackled. See Egner et al., Biophysical Journal 93:3285 (2007).

 

Allan Kachelmeier

Manager, confocal microscopy core

Oregon Hearing Research Center

OHSU

 

On 12/5/07, John Oreopoulos <[hidden email]> wrote:

Search the CONFOCAL archive at <a onclick="return top.js.OpenExtLink(window,event,this)" href="http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal" target="_blank"> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

STORM and PALM are similar. Here is the original publication on the PALM technique:

 

Betzig, E., G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess. 2006. Imaging intracellular fluorescent proteins at nanometer resolution. Science 313:1642-1645.  

 

 

John Oreopoulos, BSc,

PhD Candidate

University of Toronto

Institute For Biomaterials and Biomedical Engineering

Centre For Studies in Molecular Imaging

 

Tel: W:416-946-5022

 

 

On 5-Dec-07, at 11:04 AM, Michael Weber wrote:

Search the CONFOCAL archive at

<a onclick="return top.js.OpenExtLink(window,event,this)" href="http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal" target="_blank">http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

 

fyi (no commercial interest):

 

<a onclick="return top.js.OpenExtLink(window,event,this)" href="http://www.zeiss.de/C1256A770030BCE0/WebViewAllE/162CCB55A50A433FC12573A8002B17D9" target="_blank">http://www.zeiss.de/C1256A770030BCE0/WebViewAllE/162CCB55A50A433FC12573A8002B17D9

 

The press release contains quite some side-kicks to Leica. And the technique sounds kind of STORM-like. Does somebody has further information?

 

cheers,

Michael

 

[Chris Wood-5]

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No surprise at all that Zeiss picked up this technique, no way they would
let Leica hog the field for long. But I can't see how they can get much
out of this, as it's basically an imaging protocol, rather than an
instrumentation advance. OK, you can package up the acquisition parameters
and post-processing steps in a module/macro, but there's nothing to stop
anyone executing this technique on any microscope with the minumum of
effort. And it seems to me that STED will be suitable for live cells
sooner than the PALM-STORM techniques.

This doesn't look much more than a spoiler tactic to me on the part of
Zeiss.

Saludos

Chris

>I think you're right.  Looks as if they've teamed up with the group
>headed by Eric Betzig.  He had the paper in Science last year:
>
>"Imaging Intracellular Fluorescent Proteins at Nanometer Resolution"
>Eric Betzig, George H. Patterson, Rachid Sougrat, O. Wolf Lindwasser,
>Scott Olenych, Juan S. Bonifacino, Michael W. Davidson, Jennifer
>Lippincott-Schwartz, and Harald F. Hess
>Science 15 September 2006 313: 1642-1645
>
>If that's it, it seems like a powerful technique, lower-cost than the
>Leica system, but unfortunately also very slow.  Also, it appears only
>applicable to photoactivatable fluorescent proteins (or dyes) that will
>change their spectrum to a longer wavelength when excited with a shorter
>wavelength.  It has the advantage of using much less intense
>illumination than the Leica system but I don't know of anyone using it
>for live-cell work.
>
>Martin
>--
>Martin Wessendorf, Ph.D.                   office: [phone]
>Assoc Prof, Dept Neuroscience                 lab: [phone]
>University of Minnesota             Preferred FAX: [phone]
>6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
>Minneapolis, MN  55455             E-mail: martinw[at]med.umn.edu
>=========================================================================

[Csucs Gabor]

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It will be also interesting to see the "patent fight". A few month ago
I've listened to a talk by Stefan Hell, where he nicely presented that
actually the basis for STED (as for STORM/PALM) is the fact that dye
molecules have two switchable states - bright/dark. But this doesn't
need to be bright and dark - any system which has two distinct,
switchable states (color change or something else...) could be used to
reach high resolution. So, basically although the realization behind
STED and PALM/STROM is completely different, but the basic condition is
the same....

Cheers   Gabor

[Martin Wessendorf]

Search the CONFOCAL archive at
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Hey, Chris--

Chris Wood wrote:

> And it seems to me that STED will be suitable for live cells
> sooner than the PALM-STORM techniques.

Interesting question.  Someone pointed out to me off-list that with
STED, the fluorophore gets taken up to the excited state many more times
(--presumably 25x for a 5x increase in resolution) than with
conventional multiphoton microscopy for each photon of actual
fluorescence.  However, for each time it's in the excited state, it has
the same probability of bad photochemistry occuring--e.g., transition to
the triplet state and/or phototoxicity.  In this example, you'd expect
25x more phototoxicity with STED than with multiphoton in order to
obtain the same image.  Thus there may be inherent limitations with STED
with regards to live-cell imaging.

That being said, it'd appear that the weakness is relative: if you can
create a fluorophore that has a lower probability of engaging in bad
photochemistry, live-cell might be doable.  So it strikes me that (in
agreement with what other people have already pointed out) we'll
probably need new, better fluorophores to do live-cell imaging with STED.

Martin Wessendorf
--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455             E-mail: martinw[at]med.umn.edu