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.
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.