Main organs including heart, liver, spleen, lung, kidney, testis, and bladder were fixed in 10% formalin for 48 h and made into paraffin section.
After 15 min bladders were fixed with glutaraldehyde, filled with distilled water, and suspended in a hormone-free solution of 232 mM mannitol and 1 mM CaCl2.
Excised bladders were fixed in 4% (w/v) paraformaldehyde dissolved in 100 mM sodium cacodylate (pH 7.4) buffer for 2 h at room temperature.
C57BL/6 mouse bladders were fixed in 4% paraformaldehyde for 10 minutes and cryoprotected in 10% sucrose for 1 hour and 30% sucrose overnight.
Remaining bladder pieces were fixed in formalin and frozen sections stained with hematoxylin and eosin (H&E) to document that only epithelium was removed.
Sections of control and DMSO pretreated intact bladder dome were fixed (4% neutral buffered formalin), processed, and embedded in paraffin.
Bladder and kidney tissues were fixed in a paraformaldehyde-lysine-periodate [ 14] solution for 24 hours, then washed 3 times in sterile saline and transferred to 70% ethanol prior to processing.
The bladder was immersion fixed in 4% buffered formaldehyde and processed for paraffin sectioning.
The TRAMP mice in control or G G4-treated G G4-treatedkilled, and the entire groupsurinary tract containing seminal vesicles, prostate (including dorsal, lateral, ventral and anterior lobes) and urethra (thus excluding the urinary bladder), wereisolated, fixed and stained with hematoxylin/eosin.
Lungs, liver, kidney, urinary bladder, and all organs with suspected macroscopical lesions were fixed in 10% formalin, cut in standardized sections and stained with hematoxylin and eosin.
