Protocol for imaging C. elegans on inverted spinning disk confocal microscope

[Jay Vyas]

Hi-

We have a collaborator who is interested in confocal microscopy of C.
elegans. We have not done work with this organism and are looking for the
best way to immobilize the worm and prepare the sample for imaging on our
spinning disk confocal microscope. We have a typical inverted microscope set-
up (Nikon body).
The collaborator prefers to keep the worm(s) alive, so we need a method to
immobilize them (anesthesia?).
Do folks use agar when mounting samples? Which slides are typically used for
this purpose? If there is a reference, I would love to get the citation.

Thanks,
Jay Vyas
Massachusetts General Hospital

[Guy Cox-2]

I've never worked with C elegans but menthol is an effective anaesthetic
for many invertebrates.  Just put a lump in the medium and remove it
when they stop moving.  Rinse and dry the menthol for next time.

                                         Guy

Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
     http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Australian Centre for Microscopy & Microanalysis,
Madsen Building F09, University of Sydney, NSW 2006

Phone [phone]     Fax [phone]
             Mobile [phone]
______________________________________________
      http://www.guycox.net
 

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Jay Vyas
Sent: Thursday, 26 August 2010 2:03 AM
To: [hidden email]
Subject: Protocol for imaging C. elegans on inverted spinning disk
confocal microscope

Hi-

We have a collaborator who is interested in confocal microscopy of C.
elegans. We have not done work with this organism and are looking for
the
best way to immobilize the worm and prepare the sample for imaging on
our
spinning disk confocal microscope. We have a typical inverted microscope
set-
up (Nikon body).
The collaborator prefers to keep the worm(s) alive, so we need a method
to
immobilize them (anesthesia?).
Do folks use agar when mounting samples? Which slides are typically used
for
this purpose? If there is a reference, I would love to get the citation.

Thanks,
Jay Vyas
Massachusetts General Hospital

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01:31:00

[Julio Vazquez]

Wolke, Jezuit and Priess, 

Development 134, 2227-2236 (June 15, 2007). 

Brenner, S. (1974). The genetics of Caenorhabditis elegans. Genetics 77, 71-94.

C. elegans: Methods and Applications (Methods in Molecular Biology; Humana Press 2006) 

for example....

 

--

Julio Vazquez

Fred Hutchinson Cancer Research Center

http://www.fhcrc.org/

On Aug 25, 2010, at 9:02 AM, Jay Vyas wrote:

Hi-

We have a collaborator who is interested in confocal microscopy of C. 

elegans. We have not done work with this organism and are looking for the 

best way to immobilize the worm and prepare the sample for imaging on our 

spinning disk confocal microscope. We have a typical inverted microscope set-

up (Nikon body). 

The collaborator prefers to keep the worm(s) alive, so we need a method to 

immobilize them (anesthesia?).

Do folks use agar when mounting samples? Which slides are typically used for 

this purpose? If there is a reference, I would love to get the citation.

Thanks,

Jay Vyas

Massachusetts General Hospital

[Julio Vazquez]

See also:

Podbilewicz and Gruenbaum: Live Imaging of Caenorhabditis elegans; Chapter 20 of Live Cell Imaging, A Laboratory manual. CSHL Press. (Goldman and Spector). They describe several methods for mounting live worms.

--

Julio Vazquez, 

Fred Hutchinson Cancer Research Center

Seattle, WA 98109-1024

http://www.fhcrc.org/

On Aug 25, 2010, at 9:02 AM, Jay Vyas wrote:

Hi-

We have a collaborator who is interested in confocal microscopy of C. 

elegans. We have not done work with this organism and are looking for the 

best way to immobilize the worm and prepare the sample for imaging on our 

spinning disk confocal microscope. We have a typical inverted microscope set-

up (Nikon body). 

The collaborator prefers to keep the worm(s) alive, so we need a method to 

immobilize them (anesthesia?).

Do folks use agar when mounting samples? Which slides are typically used for 

this purpose? If there is a reference, I would love to get the citation.

Thanks,

Jay Vyas

Massachusetts General Hospital