![]() ![]() Apoptotic cells were identified by morphological changes of apoptosis such as nuclear fragmentation and condensation. ![]() After conventional hematoxylin-eosin (H&E) staining, the overall histological structure of the tumors and apoptosis were morphologically evaluated. Tumor xenografts were fixed in 10% neutral-buffered formalin, dehydrated and embedded in paraffin for slides preparation. Morphological detection of apoptosis in tumors FaDu xenografts of untreated controls, MSC (0.2mg/d × 7), irinotecan (100mg/kg/d × 7 or 14) and combination were collected 24h after last drug treatments. The VMI index was determined by image analysis using the double stained (CD31/α-SMA) frozen tissue sections (ten randomly selected microscopic fields x400 of three tumors from each group) using Analyze calculating the total number of CD31 +/α-SMA + areas and areas positive for CD31 alone as published earlier in more details. ![]() Using two chromogens (fast red and DAB), endothelial cells were immunestained red and pericytes were stained brown. Double immunohistochemical staining for endothelial cells and pericytes in tumors We have developed this method in order to determine the vascular maturation index (VMI), which is a percentage of endothelial cells (detected with MAb CD31) associated with pericytes (detected with MAb for α-smooth muscle actin, α-SMA). ![]()
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