Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear Stephen and Shalin,
I have used BioFolie in combination with a special Zeiss 1.2 NA water
lens w/o coverglass correction and with various dipping lenses. The main
advantage here was to gain the 170micron working distance (minus
30micron thickness of the transparent film) which was otherwise taken by
the coverglass.
A colleague at Toulouse Univ uses BioFolie to investigate the
cytosceleton dynamics using GFP labeled probes during Arabidopsis root
growth. It seems like the roots love to grow in between an agar pad and
BioFolie and can nicely be imaged this way.
Because there is little choice of high NA lenses without coverglass
correction, I would tend towards plastic materials with refractive
indices closer to glass like the earlier mentioned products by ibidi. On
top of good optical quality and permeabiliy for gases, those also
feature different surface treatments to improve cell adhesion
(ibi-treat, polylysine etc).
No commercial interests.
regards, jens
---
Dr. Jens Rietdorf
Head Microscopy
Novartis Research Foundation
Friedrich-Miescher-Institute, wro1066.2.32
Maulbeerstr.66, CH-4058 Basel, Switzerland
phone +41(61)69-75172 mobil +41 798284737
Email:[log in to unmask]
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Stephen Cody
Sent: Dienstag, 17. Juli 2007 01:19
To: [log in to unmask]
Subject: Re: Culture vessels for live cell confocal microscopy
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
G'day Sahlin,
Along similar lines to the Opticell that you have already mentioned you
should also consider trying "BioFolie25".
www.ivss.de/en/Products/Culture_Systems/bioFOLIE_25___petriPERM/body_bio
folie_25___petriperm.html
This will also allow CO2 and O2 to perfuse through the film. Importantly
the Refractive Index of this film is close to water at:
1.35 . So the film should be suitable for use with dipping lenses. It
can be heat welded into bags which should suit your requirements of both
upright and inverted microscopy.
I've never tried this product, so I would be interested in peoples
comments (if any).
In answer to your question: The RI of the product you mention Opticell
is 1.6 and so should not be too bad when used with either an oil lens
(if you want to image very close to the membrane) or you may be able to
use a water lens with correction collar, the slightly higher RI than
glass will to some extent offset the thin membrane. I would recommend
you use a trial and error approach, trying various collar settings and
refocusing the microscope, while examining the sharpness of the image by
eye using widefield fluorescence microscopy. With any luck the spherical
aberration correction you require may be within the range of your
collar. If not, you may find that the image quality, if not ideal, is
adequate for your research goals
No commercial affiliation with makers of BioFolie.
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: 61 3 9341 3155 Fax: 61 3 9341 3104
email: [log in to unmask]
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 Shalin Mehta
Sent: Mon 16/07/2007 4:02 PM
To: [log in to unmask]
Subject: Culture vessels for live cell confocal microscopy
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Dear List,
After a month or so, I will need to use upright and inverted confocals
with different capabilities located 1 hour bus journey apart.
I have been searching for culture vessel that allows live cell
microscopy with following features (decreasing order of preference).
1. Can be used with both inverted and upright microscopes for live cell
imaging 2. multiple chambers for multiplexing measurements 3. ease of
transportation.
I tried samples from this:
1. Nunc -- Lab-tek chambered slides - For fixed samples, it works nicely
but for live cells the thick bottom of slide won't work with objectives
on inverted microscope. It cannot be turned upside down to view adherent
cells under upright microscope. Not easy to transport.
2. ibidi -- u-slide 6 - Works with inverted scope because bottom is
coverslip-like. Can be turned upside down (for using with upright) when
wells of all channels are sealed. But then condenser cannot come close
enough because slide is resting on wells and hence cannot get good
illumination required for taking reference images. Relatively easy to
transport live cells.
3. Nunc -- Opticell - (Yet to try) appears to work with both upright and
inverted scope. Comes with special transportation envelope. Its
memberanes are 75 micron thick (less than half the coverslip thickness),
so I am concerned about errors due to wrong coverslip thickness. Lacks
multiple chambers and requires large volume of reagents.
Please share your suggestions on modifying one of this or using another
product?
Regards
Thanks
Shalin
--
My co-ordinates:
Shalin Mehta, Graduate student
Graduate Programme in Bioengineering, NUS, Singapore
Email: shalin {dot} mehta {at} gmail {dot} com
Blog: electricsbm.blogspot.com <http://electricsbm.blogspot.com/>
Mobile: +65 90694182
|
