fixation of tissues using 4% paraformaldehyde

5 posts / 0 new
Last post
kachang
kachang's picture
fixation of tissues using 4% paraformaldehyde

I am fixing rat ovary using 4% paraformaldehye for IHC.tissues are not intact but nick and crack appears around the tissues  on sectioning.what exact steps and timings should i follow.pls help

Arvind Singh Pundir
Arvind Singh Pundir's picture
 Hi,

 Hi,
the fixation time depends on thickness of your tissue, for rat ovary overnight would be enough also check the pH of PFA , clarify how you are processing your tissue after fixation means are you going for cryosectioning or paraffin sectioning if cryo then are you going for cryoprotection in 30 % sucrose befoere sectioning.............elaborate your querry for further help

kachang
kachang's picture
thank u pundir.

thank u pundir.
the ovary size is about 3cm x 3cm, PFA of pH7.4, and tissue processed in paraffin.tissue fixed for 72 hours as tissue doesnt get fixed in 24 or 48 hrs.fixed tissue are processed as follows:
PBS 1x wash - 3x -30 min each
dehydration in alcohol series of
30%,50% -30 min each,
70% -overnight,
90%,100% -30 min each,
100%:xylene-20min
xylene I & II -20 min each
Xylene : wax - 45 min
wax I & II - 45 min each
wax III - 1 hr
embedding

drgaju.15
drgaju.15's picture
hai

hai
can any body tell me 4% paraformaldehyde and 10% formalin is same or different

thanks

Matizales
Matizales's picture
Hello,

Hello,

I hope you solve your doubts.
Anyway, 4% formalin and 10% formalin is the same, as it comes from 35-37'5% and you could consider it as 4% (37%) or 10% if you add 9 times solvent.
Paraformaldehyde is the polymer and you can consider it in the same way. You will use PFA when the fixation concentration is critical.

Good luck,

Pedro