membrane for tissue culture

[Sylvie Le Guyader]

*****
To join, leave or search the confocal microscopy listserv, go to:
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*****

Dear all



One of our users needs to culture a piece of tissue on some sort of membrane. I am wondering if anyone knows where we could find such a membrane.



The requirements are:

-          It should be compatible with tissue culture (not toxic)

-          It should float (so that the top of the tissue is in the air and the bottom touches the medium)

-          It should be somehow rigid (so that it doesn't curl up)

-          It should not have any high edges like transwell inserts (because we need to image it on an upright system with a short working distance objective)



Would anyone know of something we can use?



thanks

Med vänlig hälsning / Best regards

Sylvie

@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Unit Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7,
Novum, G lift, floor 6
14157 Huddinge
Sweden
mobile: [phone]
office: [phone]
LCI website<http://ki.se/en/bionut/welcome-to-the-lci-unit>

[WILDE Geraint]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Sylvie,

I my past research days I grew organotypic hippocampal cultures which required the precise conditions you list below, and also used them so that I could image the cultures (LWD objectives at the time on an inverted scope).  I used these. http://www.merckmillipore.com/GB/en/product/Millicell-Cell-Culture-Inserts,MM_NF-C10504?bd=1 It seems they are still manufactured even after all these years.

Best

Geraint

Product Manager for Microscopy Systems

Andor Technology
Part of the Oxford Instruments Group

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Sylvie Le Guyader
Sent: 09 September 2015 08:09
To: [hidden email]
Subject: [CONFOCALMICROSCOPY] membrane for tissue culture

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear all



One of our users needs to culture a piece of tissue on some sort of membrane. I am wondering if anyone knows where we could find such a membrane.



The requirements are:

-          It should be compatible with tissue culture (not toxic)

-          It should float (so that the top of the tissue is in the air and the bottom touches the medium)

-          It should be somehow rigid (so that it doesn't curl up)

-          It should not have any high edges like transwell inserts (because we need to image it on an upright system with a short working distance objective)



Would anyone know of something we can use?



thanks

Med vänlig hälsning / Best regards

Sylvie

@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Unit Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7,
Novum, G lift, floor 6
14157 Huddinge
Sweden
mobile: [phone]
office: [phone]
LCI website<http://ki.se/en/bionut/welcome-to-the-lci-unit>


This message has been scanned for malware by SurfControl plc. www.surfcontrol.com

[Dar Do]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Sylvie,

I'm interested to have the same performances for the cell cultures I use.
Right now we are using Multiwell with Polycarbonate Membrane Transwell®
Inserts with a long range objective, but we would like to work with a short
range objective also. So we are thinking about to use Snapwell™ Inserts (
http://www.sigmaaldrich.com/catalog/product/sigma/cls3407?lang=fr&region=FR),
that give you the possibility to detach the membrane and place it on a
glass slide. My doubt are about contamination and sollicitation of the
cells, so we never try this solution so far, but I think we will in the
near future.

Best regards,

Dario

DARIO DONNARUMMA

POSTDOC
Laboratoire Charles Coulomb (L2C)

Tél : [phone] <+33+(0)+4+67+14+38+60> | Fax : [phone]
46 37

[hidden email] <[hidden email]>
<http://www.umontpellier.fr/> <http://www.umontpellier.fr/>

UNIVERSITÉ DE MONTPELLIER

CC 0026 - Place Eugène Bataillon
34095 Montpellier Cedex 5

WWW.UMONTPELLIER.FR <http://www.umontpellier.fr/>

2015-09-09 10:05 GMT+02:00 WILDE Geraint <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Sylvie,
>
> I my past research days I grew organotypic hippocampal cultures which
> required the precise conditions you list below, and also used them so that
> I could image the cultures (LWD objectives at the time on an inverted
> scope).  I used these.
> http://www.merckmillipore.com/GB/en/product/Millicell-Cell-Culture-Inserts,MM_NF-C10504?bd=1
> It seems they are still manufactured even after all these years.
>
> Best
>
> Geraint
>
> Product Manager for Microscopy Systems
>
> Andor Technology
> Part of the Oxford Instruments Group
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Sylvie Le Guyader
> Sent: 09 September 2015 08:09
> To: [hidden email]
> Subject: [CONFOCALMICROSCOPY] membrane for tissue culture
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all
>
>
>
> One of our users needs to culture a piece of tissue on some sort of
> membrane. I am wondering if anyone knows where we could find such a
> membrane.
>
>
>
> The requirements are:
>
> -          It should be compatible with tissue culture (not toxic)
>
> -          It should float (so that the top of the tissue is in the air
> and the bottom touches the medium)
>
> -          It should be somehow rigid (so that it doesn't curl up)
>
> -          It should not have any high edges like transwell inserts
> (because we need to image it on an upright system with a short working
> distance objective)
>
>
>
> Would anyone know of something we can use?
>
>
>
> thanks
>
> Med vänlig hälsning / Best regards
>
> Sylvie
>
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Unit Manager
> Karolinska Institutet- Bionut Dpt
> Hälsovägen 7,
> Novum, G lift, floor 6
> 14157 Huddinge
> Sweden
> mobile: [phone]
> office: [phone]
> LCI website<http://ki.se/en/bionut/welcome-to-the-lci-unit>
>
>
> This message has been scanned for malware by SurfControl plc.
> www.surfcontrol.com
>

[WILDE Geraint]

Sorry, I should probably have added that the transwell I referred to, if you pick the appropriate size (e.g. for a 6-well plate) would allow access for a shorter working distance objective on an upright microscope.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Dar Do
Sent: 09 September 2015 11:12
To: [hidden email]
Subject: Re: [CONFOCALMICROSCOPY] membrane for tissue culture

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Sylvie,

I'm interested to have the same performances for the cell cultures I use.
Right now we are using Multiwell with Polycarbonate Membrane Transwell® Inserts with a long range objective, but we would like to work with a short range objective also. So we are thinking about to use Snapwell™ Inserts ( http://www.sigmaaldrich.com/catalog/product/sigma/cls3407?lang=fr&region=FR),
that give you the possibility to detach the membrane and place it on a glass slide. My doubt are about contamination and sollicitation of the cells, so we never try this solution so far, but I think we will in the near future.

Best regards,

Dario

DARIO DONNARUMMA

POSTDOC
Laboratoire Charles Coulomb (L2C)

Tél : [phone] <+33+(0)+4+67+14+38+60> | Fax : [phone]
46 37

[hidden email] <[hidden email]> <http://www.umontpellier.fr/> <http://www.umontpellier.fr/>

UNIVERSITÉ DE MONTPELLIER

CC 0026 - Place Eugène Bataillon
34095 Montpellier Cedex 5

WWW.UMONTPELLIER.FR <http://www.umontpellier.fr/>

2015-09-09 10:05 GMT+02:00 WILDE Geraint <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Sylvie,
>
> I my past research days I grew organotypic hippocampal cultures which
> required the precise conditions you list below, and also used them so
> that I could image the cultures (LWD objectives at the time on an
> inverted scope).  I used these.
> http://www.merckmillipore.com/GB/en/product/Millicell-Cell-Culture-Ins
> erts,MM_NF-C10504?bd=1 It seems they are still manufactured even after
> all these years.
>
> Best
>
> Geraint
>
> Product Manager for Microscopy Systems
>
> Andor Technology
> Part of the Oxford Instruments Group
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]]
> On Behalf Of Sylvie Le Guyader
> Sent: 09 September 2015 08:09
> To: [hidden email]
> Subject: [CONFOCALMICROSCOPY] membrane for tissue culture
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all
>
>
>
> One of our users needs to culture a piece of tissue on some sort of
> membrane. I am wondering if anyone knows where we could find such a
> membrane.
>
>
>
> The requirements are:
>
> -          It should be compatible with tissue culture (not toxic)
>
> -          It should float (so that the top of the tissue is in the air
> and the bottom touches the medium)
>
> -          It should be somehow rigid (so that it doesn't curl up)
>
> -          It should not have any high edges like transwell inserts
> (because we need to image it on an upright system with a short working
> distance objective)
>
>
>
> Would anyone know of something we can use?
>
>
>
> thanks
>
> Med vänlig hälsning / Best regards
>
> Sylvie
>
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Unit Manager
> Karolinska Institutet- Bionut Dpt
> Hälsovägen 7,
> Novum, G lift, floor 6
> 14157 Huddinge
> Sweden
> mobile: [phone]
> office: [phone]
> LCI website<http://ki.se/en/bionut/welcome-to-the-lci-unit>
>
>
> This message has been scanned for malware by SurfControl plc.
> www.surfcontrol.com
>

[Sylvie Le Guyader]

Hi Geraint

The problem with the transwells you suggest is that on an upright microscope, especially with a short WD objective, the objective will bump into the wall of the well before you reach the focus plane. Or did I misunderstand? We use a Zeiss 20x water dipping lens so it is quite wide.

Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Unit Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7,
Novum, G lift, floor 6
14157 Huddinge
Sweden
mobile: [phone]
office: [phone]
LCI website


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of WILDE Geraint
Sent: den 9 september 2015 12:22
To: [hidden email]
Subject: Re: membrane for tissue culture

Sorry, I should probably have added that the transwell I referred to, if you pick the appropriate size (e.g. for a 6-well plate) would allow access for a shorter working distance objective on an upright microscope.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Dar Do
Sent: 09 September 2015 11:12
To: [hidden email]
Subject: Re: [CONFOCALMICROSCOPY] membrane for tissue culture

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Sylvie,

I'm interested to have the same performances for the cell cultures I use.
Right now we are using Multiwell with Polycarbonate Membrane Transwell® Inserts with a long range objective, but we would like to work with a short range objective also. So we are thinking about to use Snapwell™ Inserts ( http://www.sigmaaldrich.com/catalog/product/sigma/cls3407?lang=fr&region=FR),
that give you the possibility to detach the membrane and place it on a glass slide. My doubt are about contamination and sollicitation of the cells, so we never try this solution so far, but I think we will in the near future.

Best regards,

Dario

DARIO DONNARUMMA

POSTDOC
Laboratoire Charles Coulomb (L2C)

Tél : [phone] <+33+(0)+4+67+14+38+60> | Fax : [phone]
46 37

[hidden email] <[hidden email]> <http://www.umontpellier.fr/> <http://www.umontpellier.fr/>

UNIVERSITÉ DE MONTPELLIER

CC 0026 - Place Eugène Bataillon
34095 Montpellier Cedex 5

WWW.UMONTPELLIER.FR <http://www.umontpellier.fr/>

2015-09-09 10:05 GMT+02:00 WILDE Geraint <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Sylvie,
>
> I my past research days I grew organotypic hippocampal cultures which
> required the precise conditions you list below, and also used them so
> that I could image the cultures (LWD objectives at the time on an
> inverted scope).  I used these.
> http://www.merckmillipore.com/GB/en/product/Millicell-Cell-Culture-Ins
> erts,MM_NF-C10504?bd=1 It seems they are still manufactured even after
> all these years.
>
> Best
>
> Geraint
>
> Product Manager for Microscopy Systems
>
> Andor Technology
> Part of the Oxford Instruments Group
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]]
> On Behalf Of Sylvie Le Guyader
> Sent: 09 September 2015 08:09
> To: [hidden email]
> Subject: [CONFOCALMICROSCOPY] membrane for tissue culture
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all
>
>
>
> One of our users needs to culture a piece of tissue on some sort of
> membrane. I am wondering if anyone knows where we could find such a
> membrane.
>
>
>
> The requirements are:
>
> -          It should be compatible with tissue culture (not toxic)
>
> -          It should float (so that the top of the tissue is in the air
> and the bottom touches the medium)
>
> -          It should be somehow rigid (so that it doesn't curl up)
>
> -          It should not have any high edges like transwell inserts
> (because we need to image it on an upright system with a short working
> distance objective)
>
>
>
> Would anyone know of something we can use?
>
>
>
> thanks
>
> Med vänlig hälsning / Best regards
>
> Sylvie
>
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Unit Manager
> Karolinska Institutet- Bionut Dpt
> Hälsovägen 7,
> Novum, G lift, floor 6
> 14157 Huddinge
> Sweden
> mobile: [phone]
> office: [phone]
> LCI website<http://ki.se/en/bionut/welcome-to-the-lci-unit>
>
>
> This message has been scanned for malware by SurfControl plc.
> www.surfcontrol.com
>

[Sylvie Le Guyader]

Thanks to everyone for the very informative answers. These are really nice suggestions.
Jean-Marie, do you have any experience with the SimPore membranes? Are they compatible with cell culture?
And do you know if it is possible to remove the reservoirs from the Ibidi slides to allow more space for the objective?

Dario: interesting product! Thanks. There is no info on the Sigma website about the high of the walls around the membrane after it has been popped out of the insert. I said in my answer to Geraint that we use the 20x dipping lens but that was a mistake. We use a 20x dry lens NA 0.6 WD 1.7 mm. Quite a lot of WD but considering that the insert is 12mm in diameter, the walls still look too high to me. do you have any idea how high they are?

Thanks everyone! :)

Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Unit Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7,
Novum, G lift, floor 6
14157 Huddinge
Sweden
mobile: [phone]
office: [phone]
LCI website


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Dar Do
Sent: den 9 september 2015 12:12
To: [hidden email]
Subject: Re: membrane for tissue culture

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Sylvie,

I'm interested to have the same performances for the cell cultures I use.
Right now we are using Multiwell with Polycarbonate Membrane Transwell® Inserts with a long range objective, but we would like to work with a short range objective also. So we are thinking about to use Snapwell™ Inserts ( http://www.sigmaaldrich.com/catalog/product/sigma/cls3407?lang=fr&region=FR),
that give you the possibility to detach the membrane and place it on a glass slide. My doubt are about contamination and sollicitation of the cells, so we never try this solution so far, but I think we will in the near future.

Best regards,

Dario

DARIO DONNARUMMA

POSTDOC
Laboratoire Charles Coulomb (L2C)

Tél : [phone] <+33+(0)+4+67+14+38+60> | Fax : [phone]
46 37

[hidden email] <[hidden email]> <http://www.umontpellier.fr/> <http://www.umontpellier.fr/>

UNIVERSITÉ DE MONTPELLIER

CC 0026 - Place Eugène Bataillon
34095 Montpellier Cedex 5

WWW.UMONTPELLIER.FR <http://www.umontpellier.fr/>

2015-09-09 10:05 GMT+02:00 WILDE Geraint <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Sylvie,
>
> I my past research days I grew organotypic hippocampal cultures which
> required the precise conditions you list below, and also used them so
> that I could image the cultures (LWD objectives at the time on an
> inverted scope).  I used these.
> http://www.merckmillipore.com/GB/en/product/Millicell-Cell-Culture-Ins
> erts,MM_NF-C10504?bd=1 It seems they are still manufactured even after
> all these years.
>
> Best
>
> Geraint
>
> Product Manager for Microscopy Systems
>
> Andor Technology
> Part of the Oxford Instruments Group
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]]
> On Behalf Of Sylvie Le Guyader
> Sent: 09 September 2015 08:09
> To: [hidden email]
> Subject: [CONFOCALMICROSCOPY] membrane for tissue culture
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all
>
>
>
> One of our users needs to culture a piece of tissue on some sort of
> membrane. I am wondering if anyone knows where we could find such a
> membrane.
>
>
>
> The requirements are:
>
> -          It should be compatible with tissue culture (not toxic)
>
> -          It should float (so that the top of the tissue is in the air
> and the bottom touches the medium)
>
> -          It should be somehow rigid (so that it doesn't curl up)
>
> -          It should not have any high edges like transwell inserts
> (because we need to image it on an upright system with a short working
> distance objective)
>
>
>
> Would anyone know of something we can use?
>
>
>
> thanks
>
> Med vänlig hälsning / Best regards
>
> Sylvie
>
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Unit Manager
> Karolinska Institutet- Bionut Dpt
> Hälsovägen 7,
> Novum, G lift, floor 6
> 14157 Huddinge
> Sweden
> mobile: [phone]
> office: [phone]
> LCI website<http://ki.se/en/bionut/welcome-to-the-lci-unit>
>
>
> This message has been scanned for malware by SurfControl plc.
> www.surfcontrol.com
>

[Giovanna Expósito Romero]

Dear Sylvie,

You could try with MILLICELL-CM 0.4um, PICM0RG50 with 35mm dishes. They have 30mm diameter and just 5mm height.

https://www.merckmillipore.com/GB/en/product/Millicell-Cell-Culture-Insert%2C-30%C2%A0mm%2C-hydrophilic-PTFE%2C-0.4%C2%A0%C2%B5m,MM_NF-PICM0RG50#documentation

We normally use this membranes on an inverted microscope with long working distance objectives and 35mm dishes (MaTtek) for brain slice culture. I think they should work for your application also.

Best regards,

Giovanna

Giovanna Expósito
Servicio de Imagen
Instituto de Neurociencias
Telf: [phone]
Avd. Ramón y Cajal s/n
Campus de San Juan
03550 Sant Joan d´Alacant (Alicante)
Spain



 

-----Mensaje original-----
De: Confocal Microscopy List [mailto:[hidden email]] En nombre de Sylvie Le Guyader
Enviado el: miércoles, 09 de septiembre de 2015 12:30
Para: [hidden email]
Asunto: Re: membrane for tissue culture

Hi Geraint

The problem with the transwells you suggest is that on an upright microscope, especially with a short WD objective, the objective will bump into the wall of the well before you reach the focus plane. Or did I misunderstand? We use a Zeiss 20x water dipping lens so it is quite wide.

Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader, PhD
Live Cell Imaging Unit Manager
Karolinska Institutet- Bionut Dpt
Hälsovägen 7,
Novum, G lift, floor 6
14157 Huddinge
Sweden
mobile: [phone]
office: [phone]
LCI website


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of WILDE Geraint
Sent: den 9 september 2015 12:22
To: [hidden email]
Subject: Re: membrane for tissue culture

Sorry, I should probably have added that the transwell I referred to, if you pick the appropriate size (e.g. for a 6-well plate) would allow access for a shorter working distance objective on an upright microscope.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Dar Do
Sent: 09 September 2015 11:12
To: [hidden email]
Subject: Re: [CONFOCALMICROSCOPY] membrane for tissue culture

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Sylvie,

I'm interested to have the same performances for the cell cultures I use.
Right now we are using Multiwell with Polycarbonate Membrane Transwell® Inserts with a long range objective, but we would like to work with a short range objective also. So we are thinking about to use Snapwell™ Inserts ( http://www.sigmaaldrich.com/catalog/product/sigma/cls3407?lang=fr&region=FR),
that give you the possibility to detach the membrane and place it on a glass slide. My doubt are about contamination and sollicitation of the cells, so we never try this solution so far, but I think we will in the near future.

Best regards,

Dario

DARIO DONNARUMMA

POSTDOC
Laboratoire Charles Coulomb (L2C)

Tél : [phone] <+33+(0)+4+67+14+38+60> | Fax : [phone]
46 37

[hidden email] <[hidden email]> <http://www.umontpellier.fr/> <http://www.umontpellier.fr/>

UNIVERSITÉ DE MONTPELLIER

CC 0026 - Place Eugène Bataillon
34095 Montpellier Cedex 5

WWW.UMONTPELLIER.FR <http://www.umontpellier.fr/>

2015-09-09 10:05 GMT+02:00 WILDE Geraint <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Sylvie,
>
> I my past research days I grew organotypic hippocampal cultures which
> required the precise conditions you list below, and also used them so
> that I could image the cultures (LWD objectives at the time on an
> inverted scope).  I used these.
> http://www.merckmillipore.com/GB/en/product/Millicell-Cell-Culture-Ins
> erts,MM_NF-C10504?bd=1 It seems they are still manufactured even after
> all these years.
>
> Best
>
> Geraint
>
> Product Manager for Microscopy Systems
>
> Andor Technology
> Part of the Oxford Instruments Group
>
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]]
> On Behalf Of Sylvie Le Guyader
> Sent: 09 September 2015 08:09
> To: [hidden email]
> Subject: [CONFOCALMICROSCOPY] membrane for tissue culture
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all
>
>
>
> One of our users needs to culture a piece of tissue on some sort of
> membrane. I am wondering if anyone knows where we could find such a
> membrane.
>
>
>
> The requirements are:
>
> -          It should be compatible with tissue culture (not toxic)
>
> -          It should float (so that the top of the tissue is in the air
> and the bottom touches the medium)
>
> -          It should be somehow rigid (so that it doesn't curl up)
>
> -          It should not have any high edges like transwell inserts
> (because we need to image it on an upright system with a short working
> distance objective)
>
>
>
> Would anyone know of something we can use?
>
>
>
> thanks
>
> Med vänlig hälsning / Best regards
>
> Sylvie
>
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Unit Manager
> Karolinska Institutet- Bionut Dpt
> Hälsovägen 7,
> Novum, G lift, floor 6
> 14157 Huddinge
> Sweden
> mobile: [phone]
> office: [phone]
> LCI website<http://ki.se/en/bionut/welcome-to-the-lci-unit>
>
>
> This message has been scanned for malware by SurfControl plc.
> www.surfcontrol.com
>

[George McNamara]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

Read Wakabayashi et al 2007 and use their method of growing cells on the
underside of the transwell, and imaging that:

http://www.ncbi.nlm.nih.gov/pubmed/17308935

Histochem Cell Biol. 2007 May;127(5):463-72. Epub 2007 Feb 17.

Four-dimensional imaging of filter-grown polarized epithelial cells.

Wakabayashi Y(1), Chua J, Larkin JM, Lippincott-Schwartz J, Arias IM.

Author information:
(1)Cell Biology and Metabolism Branch, National Institute of Child Health and
Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
[hidden email]

Understanding how epithelial cells generate and maintain polarity and function
requires live cell imaging. In order for cells to become fully polarized, it is
necessary to grow them on a permeable membrane filter; however, the translucent
filter obstructs the microscope light path required for quantitative live cell
imaging. Alternatively, the membrane filter may be excised but this eliminates
selective access to apical and basolateral surfaces. Conversely, epithelial cells
cultured directly on glass exhibit different phenotypes and functions from filter
grown cells. Here, we describe a new method for culturing polarized epithelial
cells on a Transwell filter insert that allows superior live cell imaging with
spatial and temporal image resolution previously unachievable using conventional
methods. Cells were cultured on the underside of a filter support. Epithelial
cells grown in this inverted configuration exhibit a fully polarized
architecture, including the presence of functional tight junctions. This new
culturing system permits four-dimensional (three spatial dimension over time)
imaging of endosome and Golgi apparatus dynamics, and permits selective
manipulation of the apical and basolateral surfaces. This new technique has wide
applicability for visualization and manipulation of polarized epithelial cells.

PMID: 17308935  [PubMed - indexed for MEDLINE]



By now they may have published more such papers, and other groups may
have published and cited their work.

Enjoy,

George
p.s. you do not have to stand on your heads while imaging.



On 9/9/2015 2:08 AM, Sylvie Le Guyader wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all
>
>
>
> One of our users needs to culture a piece of tissue on some sort of membrane. I am wondering if anyone knows where we could find such a membrane.
>
>
>
> The requirements are:
>
> -          It should be compatible with tissue culture (not toxic)
>
> -          It should float (so that the top of the tissue is in the air and the bottom touches the medium)
>
> -          It should be somehow rigid (so that it doesn't curl up)
>
> -          It should not have any high edges like transwell inserts (because we need to image it on an upright system with a short working distance objective)
>
>
>
> Would anyone know of something we can use?
>
>
>
> thanks
>
> Med vänlig hälsning / Best regards
>
> Sylvie
>
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Unit Manager
> Karolinska Institutet- Bionut Dpt
> Hälsovägen 7,
> Novum, G lift, floor 6
> 14157 Huddinge
> Sweden
> mobile: [phone]
> office: [phone]
> LCI website<http://ki.se/en/bionut/welcome-to-the-lci-unit>
>
>    

--



George McNamara, Ph.D.
Single Cells Analyst
L.J.N. Cooper Lab
University of Texas M.D. Anderson Cancer Center
Houston, TX 77054
Tattletales http://works.bepress.com/gmcnamara/42

[Glen MacDonald-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Sylvie,
If you use the flexible membrane inserts, they can be removed from the polystyrene wall with a scalpel or small scissors.  Then place in a petri dish for imaging with an upright system, or in a large coverslip culture chamber to image on an inverted system.  The membrane can be returned to culture by placing onto another transwell insert.  Contamination is a concern, but we had reasonable success with brain slices.  Another option is to culture the inserts upside down.  

Regards,
Glen MacDonald
        Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
        Cellular Morphology Core
Center on Human Development and Disability
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
[phone]
[hidden email]







On Sep 9, 2015, at 3:39 AM, Sylvie Le Guyader <[hidden email]> wrote:

> Thanks to everyone for the very informative answers. These are really nice suggestions.
> Jean-Marie, do you have any experience with the SimPore membranes? Are they compatible with cell culture?
> And do you know if it is possible to remove the reservoirs from the Ibidi slides to allow more space for the objective?
>
> Dario: interesting product! Thanks. There is no info on the Sigma website about the high of the walls around the membrane after it has been popped out of the insert. I said in my answer to Geraint that we use the 20x dipping lens but that was a mistake. We use a 20x dry lens NA 0.6 WD 1.7 mm. Quite a lot of WD but considering that the insert is 12mm in diameter, the walls still look too high to me. do you have any idea how high they are?
>
> Thanks everyone! :)
>
> Med vänlig hälsning / Best regards
>  
> Sylvie
>  
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader, PhD
> Live Cell Imaging Unit Manager
> Karolinska Institutet- Bionut Dpt
> Hälsovägen 7,
> Novum, G lift, floor 6
> 14157 Huddinge
> Sweden
> mobile: [phone]
> office: [phone]
> LCI website
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Dar Do
> Sent: den 9 september 2015 12:12
> To: [hidden email]
> Subject: Re: membrane for tissue culture
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear Sylvie,
>
> I'm interested to have the same performances for the cell cultures I use.
> Right now we are using Multiwell with Polycarbonate Membrane Transwell® Inserts with a long range objective, but we would like to work with a short range objective also. So we are thinking about to use Snapwell™ Inserts ( http://www.sigmaaldrich.com/catalog/product/sigma/cls3407?lang=fr&region=FR),
> that give you the possibility to detach the membrane and place it on a glass slide. My doubt are about contamination and sollicitation of the cells, so we never try this solution so far, but I think we will in the near future.
>
> Best regards,
>
> Dario
>
> DARIO DONNARUMMA
>
> POSTDOC
> Laboratoire Charles Coulomb (L2C)
>
> Tél : [phone] <+33+(0)+4+67+14+38+60> | Fax : [phone]
> 46 37
>
> [hidden email] <[hidden email]> <http://www.umontpellier.fr/> <http://www.umontpellier.fr/>
>
> UNIVERSITÉ DE MONTPELLIER
>
> CC 0026 - Place Eugène Bataillon
> 34095 Montpellier Cedex 5
>
> WWW.UMONTPELLIER.FR <http://www.umontpellier.fr/>
>
> 2015-09-09 10:05 GMT+02:00 WILDE Geraint <[hidden email]>:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Dear Sylvie,
>>
>> I my past research days I grew organotypic hippocampal cultures which
>> required the precise conditions you list below, and also used them so
>> that I could image the cultures (LWD objectives at the time on an
>> inverted scope).  I used these.
>> http://www.merckmillipore.com/GB/en/product/Millicell-Cell-Culture-Ins
>> erts,MM_NF-C10504?bd=1 It seems they are still manufactured even after
>> all these years.
>>
>> Best
>>
>> Geraint
>>
>> Product Manager for Microscopy Systems
>>
>> Andor Technology
>> Part of the Oxford Instruments Group
>>
>> -----Original Message-----
>> From: Confocal Microscopy List
>> [mailto:[hidden email]]
>> On Behalf Of Sylvie Le Guyader
>> Sent: 09 September 2015 08:09
>> To: [hidden email]
>> Subject: [CONFOCALMICROSCOPY] membrane for tissue culture
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> Post images on http://www.imgur.com and include the link in your posting.
>> *****
>>
>> Dear all
>>
>>
>>
>> One of our users needs to culture a piece of tissue on some sort of
>> membrane. I am wondering if anyone knows where we could find such a
>> membrane.
>>
>>
>>
>> The requirements are:
>>
>> -          It should be compatible with tissue culture (not toxic)
>>
>> -          It should float (so that the top of the tissue is in the air
>> and the bottom touches the medium)
>>
>> -          It should be somehow rigid (so that it doesn't curl up)
>>
>> -          It should not have any high edges like transwell inserts
>> (because we need to image it on an upright system with a short working
>> distance objective)
>>
>>
>>
>> Would anyone know of something we can use?
>>
>>
>>
>> thanks
>>
>> Med vänlig hälsning / Best regards
>>
>> Sylvie
>>
>> @@@@@@@@@@@@@@@@@@@@@@@@
>> Sylvie Le Guyader, PhD
>> Live Cell Imaging Unit Manager
>> Karolinska Institutet- Bionut Dpt
>> Hälsovägen 7,
>> Novum, G lift, floor 6
>> 14157 Huddinge
>> Sweden
>> mobile: [phone]
>> office: [phone]
>> LCI website<http://ki.se/en/bionut/welcome-to-the-lci-unit>
>>
>>
>> This message has been scanned for malware by SurfControl plc.
>> www.surfcontrol.com
>>