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P.M.M. performed immunohistochemical optimization and staining of markers.
After antibody optimization and staining, cyclin D1 expression could be evaluated in 527of 557 tumors (94.6%) represented in the TMA.
Following antibody optimization and staining, it was possible to evaluate the expression of HMG-CoAR protein in 422 (84.4%) of the 500 tumors represented on the TMA.
Following antibody optimization and staining, PODXL expression could be evaluated in 260/270 (96.3%) tumours in cohort 1 and 316/320 (98.8%) tumours in cohort 2. In cohort 1, 137 (52.7%) tumours were negative for PODXL (score 0), 98 (37.7%) tumours displayed weak-moderate staining (score 1 2) and 25 (9.6%) tumours displayed high PODXL expression (score 3 4).
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Because the use of freshly fixed paraffin tumor has technical limitations, we took steps to minimize the impact of these concerns, including optimization of reagents and staining conditions, and use of freshly cut sections for the respective tissue microarrays.
Experimental challenges involved in this technology, such as antibody selection, sample preparation and optimization of staining conditions, have been addressed successfully in the past (Hennessy et al., 2010; Mannsperger et al., 2010b; Mircean et al., 2005; Spurrier et al., 2008).
Careful optimization of staining conditions is, of course, mandatory, as already reported (117, 118).
However, there are no published studies about optimization of staining procedures for a specific plant family or taxonomic group.
Brown seeds were cut and stained by prussian blue staining.
All tissues were embedded in paraffin, sectioned, and stained using standard H&E staining.
This PC is made of ye old wood, metals, and stained glass.
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