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Awesome Deanne!

I will look into this now. It will work nicely with my TEM and SEM analysis.

Thanks,

Shane

On 23 Jan 2013, at 2:11, Deanne Veronica Catmull <[log in to unmask]> wrote:

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> Have you thought about using BacLight live/dead stain (Molecular Probes/Invitrogen)? You will be able to determine the percentage of cells with ruptured membranes using this stain along with a good analysis package on your computer. As long as you set up all the appropriate controls you can get a good idea of the extent your antibiotic is affecting the cells. To look at specific cell structures though, I would follow the advice of others and utilize higher resolution techniques.
> 
> Kind regards,
> Deanne.
> 
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of nicoletta fucà
> Sent: Wednesday, 23 January 2013 2:49 AM
> To: [log in to unmask]
> Subject: Re: Imaging bacteria help
> 
> ________________________________
> Da: Rob
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> I'm interested as well.
> 
> 
> ________________________________
> Da: Rob Palmer <[log in to unmask]>
> A: [log in to unmask]
> Inviato: Martedì 22 Gennaio 2013 14:46
> Oggetto: Re: Imaging bacteria help
> 
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> Shane - what exactly are you trying to do?  It is unlikely that you will be able to differentiate between different mycolic acid structures using anything other than very well characterized antibodies together with immunoTEM.  If you are trying to distinguish cells that have mycolic acids from those that do not, there are probably easier ways unrelated to mycolic acids.  What do you want to do with your bacteria from broth?  Simply drying washed cells onto a slide coated with polylysine ought to do the trick.  I think you understand that these cells are very small compared to most eukaryotic cells and, depending on what exactly you want to see, EM or image-processing of confocal/digital-decon images may be the way to go.  You can contact me off-list if you'd like to discuss in detail.
> 
> On Jan 21, 2013, at 11:27 AM, Shane van Breda wrote:
> 
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>> Hi everyone,
>> 
>> I have two questions about imaging bacteria (specifically Tuberculosis 
>> or any Mycobacterium).
>> 
>> 1.) I am interested in the mycolic acids of the bacteria. Does anyone 
>> know of a specific fluorescent dye (with specific wavelengths) or any 
>> anti - mycolic acid antibodies for immunolabelling?
>> 
>> 2.) What would be the best way to fix the bacteria to slides from 
>> broth? They need to fixed and dead since they are pathogenic. But I 
>> would like to view whole cells and not sections from resin.
>> 
>> Thanks very much for your input.
>> 
>> Shane
> 
> Robert J. Palmer Jr., Ph.D.
> Microbial Receptors Unit
> Laboratory of Cell and Developmental Biology Natl Inst Dental Craniofacial Res - Natl Insts Health Bldg 30, Room 310
> 30 Convent Drive
> Bethesda MD 20892
> ph 301-594-0025
> fax 301-402-0396