TS Procure 812 - primary fixation Karnovsky's - transwells (TM - 013)

SAN:

SPEC No:

OPERATOR & STEPS:

OPERATOR & STEPS:

All solutions are exchanged above and below the membrane, up to step 10.

Karnovsky's fixative: 2.5% volume and 4% volume in 0.1Molarity (M) Sorenson's Phosphate buffer (pH 7.2, 300mosmol/kg) for at least 0h 40m 0s or overnight

0.1Molarity (M) buffer 0h 15m 0s

1% volume in buffer 1h 0m 0s

70% volume for at least 0h 20m 0s or overnight

95% volume 0h 20m 0s

100% volume 0h 20m 0s

100% volume dried with molecular sieve 0h 20m 0s

Remove membrane from transwell scaffold using a 45o scalpel and transfer to a glass vial with 100% volume

dried with molecular sieve.

50% volume 50% volume 0h 20m 0s

100% volume 0h 20m 0s

50% volume 50% volume Procure 812

100% volume Procure 8120h 20m 0s

100% volume Procure 812 1h 0m 0s

Embed & polymerise overnight in oven at 60°C , section with diamond knife as usual.