The other samples fixed in 4% paraformaldehyde
were processed using routine histological procedures, and embedded in paraffin.
We fixed whole embryos or PND1 mice for 24 hr at 4[degrees]C in 4% paraformaldehyde
, pH 7.4, followed by 20% sucrose for 24 hr at 4[degrees]C.
Bis(3-phenyl-3,4-dihydro-2H-1,3-benzoxazinyl)isopropane (BA-a) or Bisphenol-a as a standard benzoxazine monomer, paraformaldehyde
(99%), Tetrahydrofuran (THF), aniline (99%) and 3aminopropyltrimethoxysilane (98%) were supplied from Sigma Aldrich Co.
In this stage, 4-t-butyl-2-(N-piperdinomethyl)cyclohexanone was produced from 4- t-butyl-cyclohexanone, piperidine and paraformaldehyde
according to reference one.
Vaginas were removed and fixed in paraformaldehyde
and processed for subsequent histological examination.
The cells were cultured on coverslips for 24 hours and at the end of the drug treatment fixed in 4% paraformaldehyde
in phosphate buffer (pH 7.4) at room temperature for 15 min and then incubated with a permeabilizing solution (0.1% Triton X-100) for 10 min at 4[degrees]C.
The thyroid tissue was fixed with 4% paraformaldehyde
and embedded in paraffin.
Teratocytes were immediately fixed in 3% paraformaldehyde
in HEPES wash buffer for 10 min in a gentle vacuum and 2hrs at room temperature for fluorescent studies.
Surgical specimens were immersed in 4% paraformaldehyde
, decalcified with EDTA, and embedded in paraffin.
For this purpose, hydroxyl-containing benzoxazine monomer was first prepared by conventional ring-forming reaction of phenol with paraformaldehyde
in the presence of 6-amino-l-hexanol.
On PD 15, we intracardially perfused the rats with phosphate-buffered saline (PBS) containing 4 percent paraformaldehyde
. We removed the brains and postfixed them in 4 percent paraformaldehyde
for 24 h, followed by incubation in cryoprotectant.
Subsequently, 4% paraformaldehyde
(4[degrees]C; Sigma, USA) was added to each well (300 [micro]L/well) for 15 minutes, and glass coverslips were rinsed three times with PBS solution (1% triton X-100, pH 7.4) for 5 minutes each.