mouse kidney fixation
Posted 23 December 2009 - 10:21 AM
I am using perfusion-fixation with 4% PFA, with good blanching of the kidneys, cutting the kidneys into 3mm sections and incubating in 30% sucrose overnight. The specimens are then frozen in OCT in liquid nitrogen the next day, then cryosectioned at -22C at 2um sections the day after.
The tissue morphology overall looks well, with back-to-back tubules, patent vessels and open glomeruli. However, the tubular lumens are mostly closed/collapsed, which is not what I am seeing in other publications. This is a problem, because I am trying do IF to stain for a brush border protein, and other papers have shown widely patent tubular lumens and good staining.
Any thoughts would be appreciated!
Posted 26 December 2009 - 06:44 AM
Posted 30 December 2009 - 02:12 PM
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Posted 03 January 2010 - 12:57 AM
-yes, I tried using a brand new blade as well.