http://confocal-microscopy-list.275.s1.nabble.com/Imaging-bacteria-help-tp7579528p7579554.html
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> I use to work with BacLight live/dead stain for assessing bacteria
> viability but still, if bacteria flow onto the slide or coverslip
> (for inverted microscope), taking good images to use for
> quantitative analysisis is hard. As Shane said, it is necessary to
> fix bacteria onto the slides using a non destructive method. Any
> suggestions?
>
>
> ________________________________
> Da: Deanne Veronica Catmull <
[hidden email]>
> A: nicoletta fucà <
[hidden email]>;
> "
[hidden email]" <
[hidden email]>
> Inviato: Mercoledì 23 Gennaio 2013 1:11
> Oggetto: RE: Imaging bacteria help
>
> 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:
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> Sent: Wednesday, 23 January 2013 2:49 AM
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> Subject: Re: Imaging bacteria help
>
> ________________________________
> Da: Rob
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> I'm interested as well.
>
>
> ________________________________
> Da: Rob Palmer <
[hidden email]>
> A:
[hidden email]
> 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
>