Chitin staining

[Marco Büntzow]

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Hi everybody,

I'm quite new to confocal business, so I will have a lot of questions in
the near future, but here is the first:
What stainings do I need to visualize chitin in very small arthropods
like copepods? I want to get pictures of external morphology and my
animals are smaller than 500µm. Autofluorescence is not sufficient for
these small structures. At least, I didn't get good results yet.
So I really need your help. Thank you all in advance.

Marco

--
****************************************************************
Dipl. Biol. Marco Büntzow
Forschungsinstitut und Naturmuseum Senckenberg
Abt. Deutsches Zentrum für Marine Biodiversitätsforschung (DZMB)
- German Centre for Marine Biodiversity Research -
Südstrand 44
D-26382 Wilhelmshaven
Germany
Tel.: [phone]
Fax:  [phone]
e-mail: [hidden email]
****************************************************************

[Patrick Van Oostveldt]

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Dear Marco,

You can use confocal reflection microscopy to visualize external morphology.

Bye

Patrick


Quoting Marco Büntzow <[hidden email]>:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi everybody,
>
> I'm quite new to confocal business, so I will have a lot of questions
> in the near future, but here is the first:
> What stainings do I need to visualize chitin in very small  
> arthropods like copepods? I want to get pictures of external  
> morphology and my animals are smaller than 500µm. Autofluorescence  
> is not sufficient for these small structures. At least, I didn't get  
>  good results yet.
> So I really need your help. Thank you all in advance.
>
> Marco
>
> --
> ****************************************************************
> Dipl. Biol. Marco Büntzow
> Forschungsinstitut und Naturmuseum Senckenberg
> Abt. Deutsches Zentrum für Marine Biodiversitätsforschung (DZMB)
> - German Centre for Marine Biodiversity Research -
> Südstrand 44
> D-26382 Wilhelmshaven
> Germany
> Tel.: [phone]
> Fax:  [phone]
> e-mail: [hidden email]
> ****************************************************************

--
Dep. Moleculaire Biotechnologie
Coupure links 653
B 9000 GENT

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[Ole Sten Møller]

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Hello Marco,

I would guess that you could get nice results from autofluoroscence; maybe if you took some hints from this paper?

J. Michels "Confocal laser scanning microscopy: using cuticular autofluorescence for high resolution morphological imaging"
Journal of Microscopy, vol 227, pt 1 2007, pp.1-7

Just a thought, and I look forward to hearing about the results.

all the best,
Ole

---

Dr. Ole S. Möller

Wissenschaftlicher Mitarbeiter
Allgemeine und Spezielle Zoologie
Institut für Biowissenschaften, Universität Rostock
Universitätsplatz 2
D-18055 Rostock

Tel: [phone]
Funk: ([phone]

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[hidden email]
[hidden email]

http://www.biologie.uni-rostock.de/zoologie/moeller.htm




-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Marco Büntzow
Sent: 19. oktober 2007 15:09
To: [hidden email]
Subject: Chitin staining

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi everybody,

I'm quite new to confocal business, so I will have a lot of questions in the near future, but here is the first:
What stainings do I need to visualize chitin in very small arthropods like copepods? I want to get pictures of external morphology and my animals are smaller than 500µm. Autofluorescence is not sufficient for these small structures. At least, I didn't get good results yet.
So I really need your help. Thank you all in advance.

Marco

--
****************************************************************
Dipl. Biol. Marco Büntzow
Forschungsinstitut und Naturmuseum Senckenberg Abt. Deutsches Zentrum für Marine Biodiversitätsforschung (DZMB)
- German Centre for Marine Biodiversity Research - Südstrand 44
D-26382 Wilhelmshaven
Germany
Tel.: [phone]
Fax:  [phone]
e-mail: [hidden email]
****************************************************************

[Rosemary.White]

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http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi Marco,

We use wheat germ agglutinin to stain fungal chitin, maybe you could use it
for insect chitin as well?  The Alexa-tagged WGAs are brightest, though most
expensive, available from Molecular Probes.  You just dilute in water, and
add to your specimen for 10 min to a couple of hours, rinse, observe.
Surface waxes might block access to the chitin, I guess.  Might be worth a
try.

There are probably other non-specific stains, maybe propidium iodide or
Schiff's reagent might work on insects as well?

cheers,
Rosemary

Dr Rosemary White               [hidden email]
CSIRO Plant Industry            ph.     61 (0)2-6246 5475
GPO Box 1600                       fax.     [phone]
Canberra, ACT 2601
Australia

On 19/10/07 11:09 PM, "Marco Büntzow" <[hidden email]> wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi everybody,
>
> I'm quite new to confocal business, so I will have a lot of questions in
> the near future, but here is the first:
> What stainings do I need to visualize chitin in very small arthropods
> like copepods? I want to get pictures of external morphology and my
> animals are smaller than 500µm. Autofluorescence is not sufficient for
> these small structures. At least, I didn't get good results yet.
> So I really need your help. Thank you all in advance.
>
> Marco