🚀 go-pugleaf

Hi Gerhard,
I remember a protocoll using Nano Orange from Molecular probes published
by H.P. Grossard and F. Azam. Never tried it myself. Hope it helps

Wolfgang

gerhard schrieb:
> 
> Hi everyone
> 
> I would like to detect the presence of flagella on bacteria,
> preferentially by fluorescence.
> 
> Does anyone know of a flagellum specific fluorescent stain, perhaps an
> antibody that recognises flagella of a variety of species?
> 
> Thanks in advance
> 
> Gerhard Nebe-von-Caron

I Know that I have seen a kit for flagellar staining, but not sure where to
get it. It was not fluoresence based, but worked fairly well under a normal
scope.

Richard Oeffner
Cal State San Marcos

Wolfgang Wenzel wrote:

> Hi Gerhard,
> I remember a protocoll using Nano Orange from Molecular probes published
> by H.P. Grossard and F. Azam. Never tried it myself. Hope it helps
>
> Wolfgang
>
> gerhard schrieb:
> >
> > Hi everyone
> >
> > I would like to detect the presence of flagella on bacteria,
> > preferentially by fluorescence.
> >
> > Does anyone know of a flagellum specific fluorescent stain, perhaps an
> > antibody that recognises flagella of a variety of species?
> >
> > Thanks in advance
> >
> > Gerhard Nebe-von-Caron

I don't know of any fluorescent staining methods, but I do remember that:
1) in many cases flagella can be seen under polarized light - if the bugs
are too motile, you can try making a slide with 0.7% agar
2) careful heat fixation  (or fixation in Os vapors) and staining with
protein-binding dyes like crystal violet allows to see flagella, too.

Otherwise check Methods of Mol. Bacteriology, Ed. Gerhardt (don't remember
year, might be wrong with the title, too, but you can find it in
Amazon.com).

Emir


"gerhard" <gerne1@hotmail.com> wrote in message
news:4005dc5d.0109071943.2d7d4c24@posting.google.com...
> Hi everyone
>
> I would like to detect the presence of flagella on bacteria,
> preferentially by fluorescence.
>
> Does anyone know of a flagellum specific fluorescent stain, perhaps an
> antibody that recognises flagella of a variety of species?
>
> Thanks in advance
>
> Gerhard Nebe-von-Caron

Isn't acid fast staining ok for flagella
KC

"Emir Khatipov" <khatipovNO@NOuchicago.edu> wrote in message
news:VCoB7.130$N4.4377@news.uchicago.edu...
> I don't know of any fluorescent staining methods, but I do remember that:
> 1) in many cases flagella can be seen under polarized light - if the bugs
> are too motile, you can try making a slide with 0.7% agar
> 2) careful heat fixation  (or fixation in Os vapors) and staining with
> protein-binding dyes like crystal violet allows to see flagella, too.
>
> Otherwise check Methods of Mol. Bacteriology, Ed. Gerhardt (don't remember
> year, might be wrong with the title, too, but you can find it in
> Amazon.com).
>
> Emir
>
>
> "gerhard" <gerne1@hotmail.com> wrote in message
> news:4005dc5d.0109071943.2d7d4c24@posting.google.com...
> > Hi everyone
> >
> > I would like to detect the presence of flagella on bacteria,
> > preferentially by fluorescence.
> >
> > Does anyone know of a flagellum specific fluorescent stain, perhaps an
> > antibody that recognises flagella of a variety of species?
> >
> > Thanks in advance
> >
> > Gerhard Nebe-von-Caron
>
>

Hello, I missed the original post, but check out:
Real-Time Imaging of Fluorescent Flagellar Filaments Turner et al. J.
Bacteriol. 182:2793 (2000)
keep your wallet handy: the dyes they used aren't cheap.
cheers,
David

"Emir Khatipov" <khatipovNO@NOuchicago.edu> wrote in message
news:VCoB7.130$N4.4377@news.uchicago.edu...
> I don't know of any fluorescent staining methods, but I do remember that:
> 1) in many cases flagella can be seen under polarized light - if the bugs
> are too motile, you can try making a slide with 0.7% agar
> 2) careful heat fixation  (or fixation in Os vapors) and staining with
> protein-binding dyes like crystal violet allows to see flagella, too.
>
> Otherwise check Methods of Mol. Bacteriology, Ed. Gerhardt (don't remember
> year, might be wrong with the title, too, but you can find it in
> Amazon.com).
>
> Emir
>
>
> "gerhard" <gerne1@hotmail.com> wrote in message
> news:4005dc5d.0109071943.2d7d4c24@posting.google.com...
> > Hi everyone
> >
> > I would like to detect the presence of flagella on bacteria,
> > preferentially by fluorescence.
> >
> > Does anyone know of a flagellum specific fluorescent stain, perhaps an
> > antibody that recognises flagella of a variety of species?
> >
> > Thanks in advance
> >
> > Gerhard Nebe-von-Caron
>
>

Liefson's (?spelling) flagella stain is a fuschin based stain and is VERY
difficult to get a good stain. The glass slides must be super clean, the
organisms must be a young culture and diluted to the right concentration.
Any agitation in the dilution process or even in the smearing can dislodge
the flagella.


-- 
John Gentile MS M(ASCP)
Laboratory Information, QA Manager
VA Medical Center
Providence, RI
yjgent@home.com



> From: "Kevin Carter" <K.Carter@bham.ac.uk>
> Organization: The University of Birmingham news server
> Newsgroups: 
> 
sci.bio.microbiology,bionet.microbiology,sci.bio.immunocytochem,sci.bio.ecol
og> y
> Date: Wed, 24 Oct 2001 13:52:17 +0100
> Subject: Re: Staining of flagella
> 
> Isn't acid fast staining ok for flagella
> KC
> 
> "Emir Khatipov" <khatipovNO@NOuchicago.edu> wrote in message
> news:VCoB7.130$N4.4377@news.uchicago.edu...
>> I don't know of any fluorescent staining methods, but I do remember that:
>> 1) in many cases flagella can be seen under polarized light - if the bugs
>> are too motile, you can try making a slide with 0.7% agar
>> 2) careful heat fixation  (or fixation in Os vapors) and staining with
>> protein-binding dyes like crystal violet allows to see flagella, too.
>> 
>> Otherwise check Methods of Mol. Bacteriology, Ed. Gerhardt (don't remember
>> year, might be wrong with the title, too, but you can find it in
>> Amazon.com).
>> 
>> Emir
>> 
>> 
>> "gerhard" <gerne1@hotmail.com> wrote in message
>> news:4005dc5d.0109071943.2d7d4c24@posting.google.com...
>>> Hi everyone
>>> 
>>> I would like to detect the presence of flagella on bacteria,
>>> preferentially by fluorescence.
>>> 
>>> Does anyone know of a flagellum specific fluorescent stain, perhaps an
>>> antibody that recognises flagella of a variety of species?
>>> 
>>> Thanks in advance
>>> 
>>> Gerhard Nebe-von-Caron
>> 
>> 
> 
> 

Thanks for all your input. I had already come across the staining
methods described. However, what I was after was a stain that
specifically renders the flagellum fluorescent but not the rest of the
cell. The use of Alexa comes closest to that as it basically labels
the entire cell surface. However, I would like the cell body to remain
dark in order to investigate the cells using flow cytometry. Perhaps
the ideal would be an anti-flagellum antibody that reacts to native
flagellum - not denatured.

Thanks again for all your help


Gerhard Nebe-von-Caron

John Gentile <yjgent@home.com> wrote in message news:<B7FF81AC.231%yjgent@home.com>...
> Liefson's (?spelling) flagella stain is a fuschin based stain and is VERY
> difficult to get a good stain. The glass slides must be super clean, the
> organisms must be a young culture and diluted to the right concentration.
> Any agitation in the dilution process or even in the smearing can dislodge
> the flagella.
> 
> 
> -- 
> John Gentile MS M(ASCP)
> Laboratory Information, QA Manager
> VA Medical Center
> Providence, RI
> yjgent@home.com
> 
> 
> 
> > From: "Kevin Carter" <K.Carter@bham.ac.uk>
> > Organization: The University of Birmingham news server
> > Newsgroups: 
> > 
> sci.bio.microbiology,bionet.microbiology,sci.bio.immunocytochem,sci.bio.ecol
> og> y
> > Date: Wed, 24 Oct 2001 13:52:17 +0100
> > Subject: Re: Staining of flagella
> > 
> > Isn't acid fast staining ok for flagella
> > KC
> > 
> > "Emir Khatipov" <khatipovNO@NOuchicago.edu> wrote in message
> > news:VCoB7.130$N4.4377@news.uchicago.edu...
> >> I don't know of any fluorescent staining methods, but I do remember that:
> >> 1) in many cases flagella can be seen under polarized light - if the bugs
> >> are too motile, you can try making a slide with 0.7% agar
> >> 2) careful heat fixation  (or fixation in Os vapors) and staining with
> >> protein-binding dyes like crystal violet allows to see flagella, too.
> >> 
> >> Otherwise check Methods of Mol. Bacteriology, Ed. Gerhardt (don't remember
> >> year, might be wrong with the title, too, but you can find it in
> >> Amazon.com).
> >> 
> >> Emir
> >> 
> >> 
> >> "gerhard" <gerne1@hotmail.com> wrote in message
> >> news:4005dc5d.0109071943.2d7d4c24@posting.google.com...
> >>> Hi everyone
> >>> 
> >>> I would like to detect the presence of flagella on bacteria,
> >>> preferentially by fluorescence.
> >>> 
> >>> Does anyone know of a flagellum specific fluorescent stain, perhaps an
> >>> antibody that recognises flagella of a variety of species?
> >>> 
> >>> Thanks in advance
> >>> 
> >>> Gerhard Nebe-von-Caron
> >> 
> >> 
> > 
> >