Materials and Methods:
Tissue microarray (TMA) consisting of three to six representative 0.6-mm cores from formalin-fixed, paraffin-embedded tissue blocks was generated in the Department of Pathology and Laboratory Medicine at OSU. Glioma tissue samples were collected from patients who underwent surgery at OSU Medical Center. Deidentified clinical data is attached to each sample (e.g. age, sex, location, prior treatment, patient survival, and so on). After immunohistochemistry, tissues too small and/or crushed were eliminated, and 91 samples were processed to the quantitative analysis using Immunoratio software (http://imtmicroscope.uta.fi/immunoratio/).
Example 1: Gu C et al., Stem Cells 2013
Upper panels: graph indicating the proportion of MELK (.) cells, determined by immunohistochemistry, in primary (newly diagnosed and recurrent GBM tissues in unmatched (left) and matched (right) cases. For unmatched cases p < .0086 and for matched cases p . .038. Lower panels: representativeimages for immunohistochemistry with two matched patient samples with primary and recurrent GBM tumors. The arrows indicate vascular sclerosis by radiation-induced damage in recurrent tumors. Bars . 50 lm. Abbreviations: GBM, glioblastoma multiforme; GFP, green fluorescentprotein; MELK, maternal embryonic leucine-zipper kinase.
Example 2: Guvenc et al., Clinical Cancer Research 2012
Recurrent GBM tissues exhibit higher Survivin expression than nontreated tumors and Survivin expression is predictive of postsurgical patient survival. A, representative pictures of Survivin immunohistochemistry analyses of newly diagnosed (nontreated) and recurrent GBM tissues. Original magnification of the top is_20 and the bottom is_40. Scale bars in top and bottom panels are 50 and 20 mm, respectively. Quantified immunostaining results with newly diagnosed and recurrent GBM tissues in unmatched cases (n . 75), and primary and recurrent tumors in 10 matched GBM cases. The average of the proportion of Survivin (.) cells in each sample set is represented by the red horizontal bar (P < 0.05; right). B, a Kaplan–Meier curve depicting the survival of GBM patients with high and low levels of Survivin expression. Exact P value is indicated in the figure.