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    CLEARING AND STAINING

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    miko
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    Topic CLEARING AND STAINING

    Post by miko on Thu May 13 2010, 08:03

    I will start to post some extremely interesting pictures of some piranhas that have gone through the process of clearing and staining. This process helps ichthyologists study the bone, cartilage and nerve structures of fish.

    Here is an example on a Pygocentrus cariba:



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    CombiChrist
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    Topic Re: CLEARING AND STAINING

    Post by CombiChrist on Thu May 13 2010, 14:21

    Holy shit, wonderful pics Ivan cheers

    What is the "clearing and staining proces", do you do it yourself ?
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    Topic Re: CLEARING AND STAINING

    Post by ANDONI on Sat May 22 2010, 07:12

    CombiChrist wrote:Holy shit, wonderful pics Ivan cheers

    What is the "clearing and staining process", do you do it yourself ?

    Yes I would also like to know what this process is also.






    _______________________________________________________________________________
    Serrasalmus Marginatus X 2
    Serrasalmus Compressus
    Serrasalmus Manueli
    Pygocentrus Nattereri x 5
    Serrasalnus Rhombeus ?



    PIRANHAFEAR ON YOUTUBE

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    Topic Re: CLEARING AND STAINING

    Post by CombiChrist on Sat May 22 2010, 13:27

    Sure, I would love to try it myself. So far I didn't really succeed in the boiling to get myself a preserved bone structure, so maybe this method might be a better one Smile
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    Topic diaphanisation or clearing and staining Fish

    Post by miko on Sun May 23 2010, 00:50

    I have a video explaining the whole process but I cant upload it. My internet got to slow this month in Venezuela! Ill upload it asap.

    There are many sites that explain the whole process. It’s actually quite easy and fast if you have all the chemicals.

    Try googling: diaphanisation or clearing and staining Fish.

    Ivan
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    Topic Re: CLEARING AND STAINING

    Post by CombiChrist on Sun May 23 2010, 02:08

    Thanks for the advise and I hope you'll be able to upload soon Smile
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    Topic Re: CLEARING AND STAINING

    Post by ANDONI on Mon May 24 2010, 08:13

    That would be great if we can upload soon






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    Topic Re: CLEARING AND STAINING

    Post by ANDONI on Sat Jun 12 2010, 17:58

    Can you please explain the CLEARING AND STAINING process. The video would also help out, thanks Ivan













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    Topic Re: CLEARING AND STAINING

    Post by CombiChrist on Sat Jun 12 2010, 18:42

    No video, but the procedure :


    Procedure:

    1. Fix fresh material in 10% formalin
    2-3 days
    or longer. Specimens already in formalin or alcohol,
    ignore
    step 1 and go to step 2.

    2. Wash in several changes of distilled
    water
    for 1-2 days.

    3. Place directly into a mixture of 10
    mg Alcian
    Blue 8GN (supplied by Matheson, Coleman and Bell), 80
    ml 95%
    ethyl alcohol and 20 ml glacial acetic acid 10-12
    hours. (This
    solution must be fresh)


    4. Transfer to two changes of 95% ethyl
    alcohol,
    1-2 hours per change.

    5. Transfer through an alcohol series of
    75%,
    40%, 15% ethyl alcohol 2-3 hours in each, or until
    specimen
    sinks.

    6. Transfer to distilled water 2-3
    hours, or until
    specimen sinks

    7. Place in an enzyme solution of 30 ml
    saturated
    aqueous sodium borate, 70 ml distilled water, and 1 g
    trypsin
    enzyme. Change solution every 2-3 days, or sooner if
    solution
    turns a bluish color. Continue until bones and
    cartilage are
    clearly visible, and the flesh retains no blue color.
    NOTE:
    the skin of some gobies remains blue at this stage.

    8. If bone is present: Transfer
    to 0.5%
    aqueous KOH solution, to which enough Alizarin Red S
    stain has
    been added to turn solution a deep purple-red. Leave
    24 hours
    or until bones are distinctly red. If bone is not
    present
    go to step 9


    9. Transfer through a 0.5% KOH -
    glycerin series
    of 3/4 KOH - 1/4 glycerin, 1/2 KOH - 1/2 glycerin, 1/4
    KOH -
    3/4 glycerin, and finally to pure glycerin.

    10. Store specimen in pure glycerin to
    which a
    few crystals of thymol have been added to inhibit
    molds and
    bacteria.

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    Topic Re: CLEARING AND STAINING

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      Current date/time is Tue Jul 25 2017, 21:22