Chondrocytes

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Raul
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Joined: Tue May 12, 2015 4:55 pm
Location: Bradford, England

Chondrocytes

#1 Post by Raul » Wed Apr 13, 2016 1:54 pm

Cartilage showing chondrocytes. TBO stained. Mounted in Nail Polish.
Hope you enjoy.
800x magnifiction
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800x and 4x camera zoom

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billbillt
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Re: Chondrocytes

#2 Post by billbillt » Wed Apr 13, 2016 2:52 pm

these turned out very well..

BillT

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Raul
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Re: Chondrocytes

#3 Post by Raul » Wed Apr 13, 2016 3:11 pm

Thank you BillT. I'm glad you appreciate them.

billbillt
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Re: Chondrocytes

#4 Post by billbillt » Wed Apr 13, 2016 3:59 pm

Hi Raul,
I also am a fan of using nail polish as a mountant... So far, I have had none of the trouble posted by others... I am always looking for ways to use what I have around the house instead of buying something expensive to do the same job...

The Best,
BillT

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billben74
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Re: Chondrocytes

#5 Post by billben74 » Wed Apr 13, 2016 8:27 pm

Great images of Chondrocytes.
Any details of your processesing?
I'm interested in learning / trying such things.

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Raul
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Re: Chondrocytes

#6 Post by Raul » Wed Apr 13, 2016 10:17 pm

BillT I always try to use things that are handy, that's what an amateur is like, reinventing a purpose for some items, but it has some limitations and could lead to bad results where they could have been good.
Thank you billben74, well the whole procedure is the following:
1. Collected a chicken bone (from a chicken drumstick) and remove with a knife the epiphysis
2. Decalcify the sample in 3M citric acid (you can get this as limescale remover, get the crystals), i'm using citric acid because the citrate ion acts as a chelating agent and does not harm the tissue as much as other acids like HCl, this was done in 3 weeks period
3. Fixed the sample in 70%ethanol and 10% acetic acid for 2 days (depending on sample size, the bigger the more time is required
4.IPA 70% 6hours
5.IPA 90% 6 hours
6.IPA 100% 6 hours
8. then embed in wax 2 hours
Now for the sectioning I made myself a microtome as seen in the figures, and by turning the nut I can cut some really good sections, but it took a lot of practice and samples (but I like to think that now I have some skill at it) the blade I used is a hobby knife.
9.after sectioning I dewaxed the sample in white spirit for 6 hours (I wanted to be sure that is dewaxed and I also had the time to spare)
10. Rehydrated the sample in regressive alcohol baths.
11. Stained the tissue with 0.5% aqueous TBO for 2 min
12. Washed
13. 1% HCL, 70% ethanol wash for 20 sec
14. Washed
15. Dehydrated in progressive alcohol baths
16. Transferred to Nail Polish

That's my protocol that I used, please do tell if you need any more details.
And thank you very much for enjoying it.
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Raul
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Re: Chondrocytes

#7 Post by Raul » Thu Apr 14, 2016 11:12 pm

Yes it is a long process Lymia, but it seems to do the job :) .

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billben74
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Re: Chondrocytes

#8 Post by billben74 » Sun Apr 17, 2016 4:28 pm

Thanks very much for your protocol.
Thats perfectly clear to me.
I've only done plants for far, so its really interesting to hear about animal tissue. :)

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Raul
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Re: Chondrocytes

#9 Post by Raul » Mon Apr 18, 2016 5:15 pm

billben74 wrote:Thanks very much for your protocol.
Thats perfectly clear to me.
I've only done plants for far, so its really interesting to hear about animal tissue. :)
You're welcome :).

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