RE: [Histonet] Zebrafish histology

From:"Keller, Charles"

microCT-based virtual histology works pretty well for this purpose, too.  Resolution is limited to 1 um.  See this image at as well as others at .  My disclaimer is that I co-founded the company that does this imaging.  I'd be happy to do it in my academic lab, though, at no cost.  


Charles Keller, MD
Assistant Professor, Department of Cellular & Structural Biology
Adjunct Assistant Professor, Department of Pediatrics
Director, Small Animal Imaging Facility
Greehey Children's Cancer Research Institute
The University of Texas Health Science Center
8403 Floyd Curl Drive, Mail Code 7784
San Antonio, TX 78229-3900
210-562-9062 [office]
210-562-9014 [fax] or
Postdoctoral Opportunities and 
Undergraduate & Summer Internships:

-----Original Message-----
From: [] On Behalf Of
Sent: Thursday, February 28, 2008 11:01 AM
To: Dickey, Coral
Subject: Re: [Histonet] Zebrafish histology

Another technique that we've used to clear fish (in our case medaka) is
to clear into immersion oil but instead of oil with a refractice index
of 1.515 (typical immersion oil) we found much better results using oil
with a refractive index of 1.56.  You can get the oil from Cargille.  To
clear, pass the fish from fixation (aqueous) to methanol then to
propylene oxide then to immersion oil.  I've never tried it with the
skeletal stains.  I can give you a more detailed protocol if interested.

Kevin Flynn
Mid-Continent Ecological Division
6201 Congdon Blvd
Duluth MN 55804

             Rene J Buesa                                               
             Sent by:                 "Dickey, Coral"                   
             histonet-bounces         ,      
             @lists.utsouthwe         "histonet@lists.utsouthwestern.ed 
             02/28/2008 10:20                                        cc 
                                      Re: [Histonet] Zebrafish          

If you want to able to see the internal organs and the skeleton, why
don't completely make the fish transparent?
  To do that dehydrate in ethanol of increasing strengths, slowly and
after the fish are dehydrated, clear them with cedar wood oil. It will
take some time but you will end with fish completely cleared.
  René J.

"Dickey, Coral"  wrote:
  Hello out there,

I am doing double skeletal staining on zebrafish and we run into a
problem with visualization of th skeleton. The scales take up the
alizarin red stain and they are incredibly persistent and don't come off
the fish very well even after weeks in KOH/Glycerin solution. They make
it difficult to see the inside of the fish. The rest of the fish is
quite delicate after this length of time however, posing a challenge to
remove the scales without breaking the fish to pieces!

The first time I performed this stain, I used a plastic pipette to
debride the scales. This was incredibly tedious and time consuming. Does
anyone have experience with this or even have a good idea?

Any suggestions appreciated!
Thanks in advance,

Coral Dickey
Histology Specialist I

Stowers Institute for Medical Research
1000 E. 50th Street
Kansas City, Missouri 64110
Phone: 816-926-4305

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