Hi Mike

I can vouch for the agarose method and have taken good confocal images this way
of live immobilised bacterial cells.  Ensure you prepare the agarose slides on
a level bench just before use as specified by Heather below

Kind regards
Jen

Quoting Heather A Owen <[log in to unmask]>:

> Hi Mike,
> 
> A trick taught to me my a microbiologist (Mark McBride) is to make a 0.7%
> agarose solution in water (microwave to dissolve), and while it is still hot
> use a Pasteur pipet to flow a layer onto a microscope slide to cover it.  I
> usully suspend the slides on wooden applicator sticks in case the agarose
> runs off, otherwise the solution wicks under the slide.  Once the agarose
> gels, apply a drop of culture on top, then coverslip it.  The pad of agarose
> needs to be larger than the coverslip you want to use.  The cells stay
> immobile, and actually stay alive for a long time.
> 
> Sincerely yours,
> Heather Owen
> 
> 
> Dr. Heather A. Owen
> Director, Electron Microscope Laboratory
> Department of Biological Sciences
> University of Wisconsin - Milwaukee
> Lapham Hall
> 3209 N. Maryland Avenue
> Milwaukee, WI   53211
> (414)229-6816
> 
> ----- Original Message -----
> From: "Mike Tighe" <[log in to unmask]>
> To: [log in to unmask]
> Sent: Thursday, October 22, 2009 1:11:00 PM GMT -06:00 US/Canada Central
> Subject: Immobilize bacteria
> 
> Does anybody have a good way to Immobilize bacteria without killing. We would
> like to do a Live/dead stain and our bugs like to float away!
> 
> Thanks!!
> Mike
> 


-- 
Jennifer Clarke BSc (Hons) PhD
Research Associate, Anatomy and Histology
Centre for Neuroscience, School of Medicine
&
Facility Manager, Optical Microscopy Suite, Flinders Microscopy 
(available for training and assistance on Mondays only)

Flinders University
GPO Box 2100, Adelaide 5001
Phone: 61 8 8204 6454 / 61 8 8204 4205
Email: [log in to unmask]