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Mice were deeply anesthetized and perfused transcardially with saline followed by 4% paraformaldehyde
Bis(3-phenyl-3,4-dihydro-2H-1,3-benzoxazinyl)isopropane or Bisphenol-A as a standard benzoxazine monomer, paraformaldehyde
(99%), Tetrahydrofuran (THF), aniline (99%) and 3-aminopropyltrimethoxysilane (98%) were obtained from Sigma Aldrich Co.
SOE treated cells were fixed with 4% paraformaldehyde
solution for 15 min at 4[degrees]C.
We washed the surfaces, immediately fixed the remaining cells with 4% paraformaldehyde
, and stained the cell membranes with octadecyl rhodamine B (Invitrogen) or CellMask Deep Red (Invitrogen) according to manufacturer's instructions and the cell nuclei with DAPI (Vectashield Mounting Media, Vector Laboratories).
For DAPI staining, the scaffolds were fixed with 4% Paraformaldehyde
in PBS for 20 mins.
Tissue preparation: Bone samples were immersed in PLP fixative (2% paraformaldehyde
For immunofluorescence studies, the OC exptants were fixed in 4% paraformaldehyde
in phosphate-buffered saline (PBS) and permeabilized with 5% Triton X-100 in PBS with 10% fetal calf serum.
res] by Western blot or fixed in buffered 4% paraformaldehyde
for histopathologic studies.
The hardener was a 1:1 mixture of paraformaldehyde
and wood powder used in 20 to 100 parts by mass, based on liquid resin.
Cells were then fixed by 3% paraformaldehyde
, permeabilized and probed with antibody against [alpha.
The nerve segments were fixed in buffered 2 - 4% paraformaldehyde
at 4[degrees] C overnight.