Postinfection Eye Tissue Fixation

Whole eyes were harvested at 0, 4, 8, or 12 h postinfection and incubated in an 85/15 paraformaldehyde/alcohol fixative for 24 h at room temperature. Whole eyes were then exchanged into 70% ethanol for 24 h and then embedded in paraffin. Sections were deparaffinized and stained in Harris hematoxylin solution for 8 min, counterstained in eosin-phloxine B solution for 30 sec, dehydrated through two changes of 95% alcohol and cleared in two changes of xylene. Images are representative of 4 eyes per group at each time point.

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Novosad B.D., Astley R.A, & Callegan M.C. (2011). Role of Toll-Like Receptor (TLR) 2 in Experimental Bacillus cereus Endophthalmitis. PLoS ONE, 6(12), e28619.

Publication 2011

Dehydrated alcohol Embedded paraffin Eosin Eyes Fixative Paraformaldehyde Phloxine Phloxine b Xylene

Corresponding Organization : Dean McGee Eye Institute

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Protocol cited in 6 other protocols

Variable analysis

independent variables

Time postinfection (0, 4, 8, or 12 h)

dependent variables

Histological changes in the whole eye

control variables

Fixation of whole eyes in 85/15 paraformaldehyde/alcohol for 24 h at room temperature
Exchange of whole eyes into 70% ethanol for 24 h
Embedding of whole eyes in paraffin
Deparaffinization and staining of sections with Harris hematoxylin solution for 8 min
Counterstaining of sections with eosin-phloxine B solution for 30 sec
Dehydration of sections through two changes of 95% alcohol
Clearing of sections in two changes of xylene

controls

Positive control: Not specified
Negative control: Not specified

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