Ten microliters of the 150 μL fractions was used for conventional SDS-PAGE analysis and silver stain visualization.
Type II collagen was immunolocalized by the immunoperoxidase ABC technique (Vector, Burlingame, CA, USA), applying monoclonal primary antibodies ms. anti collagen II, clone II-4C11 (Calbiochem-Merck, Schwalbach, Germany), biotin conjugated polyclonal secondary antibodies (goat anti-mouse IgG (Jackson, West Grove, PA, USA)), and the nickel and cobalt enhanced DAB stain visualization.
In preliminary studies, individual sections of a limited sample of patient PCa tumor tissues were stained for CD147 and by IHC to optimize staining, visualization, and quantification of CD147 and E-cadherin.
Tissue sections were stained with Masson's trichrome staining for visualization of collagen, CD31 (PECAM) staining for visualization of tumor vasculature, or Ki-67 staining for mitotic index.
Staining of the UAMS-1 biofilm with the LIVE/DEAD BacLight viability stain and visualization by CLSM revealed that the tower structures arose from and were surrounded by a flat mat or "lawn" of densely packed cells that was approximately 10 µm thick.
The PCR samples were run on 2% pre-cast agarose E-gels (Life Technologies) containing a fluorescent stain for visualization.
For staining and visualization of whole skeletons, fetuses or mice were dissected and stained with alizarin red S or alcian blue 8GX (Sigma), or both, as described previously [32].
On day 14, bone wafers were TRAP-stained for OC quantification, followed by toluidine blue staining for visualization of erosion pits on the bone surface.
In each type of colitis, NBD-PZ revealed the destruction of the crypts as the disappearance of the ring-shaped fluorescent stains and the infiltration of inflammatory cells as the aggregation of punctate fluorescent stains through visualization of lysosomes.
Double-labeling detections were achieved on deparaffinized and hydrated sections by performing DAB-stained DING protein immunochemistry (brown staining) as described above, followed by GFAP or NeuN immunolabeling using SG vector dye (blue staining) for visualization [29].
The general steps for each of these markers involves pre-analytic (fixation, embedding, processing and sectioning), analytic (permeation, staining and visualization) and post-analytic steps (interpretation and diagnosis).
