To gain insight into how nucleosomes are arranged in vivo, we combined quantitative super-resolution nanoscopy with computer simulations to visualize and count nucleosomes along the chromatin fiber in single nuclei.
After the final tumor imaging, the tissues were removed and the animals were re-imaged to visualize and count cancer cell dissemination and metastases.
After 72 hours, the overlay was removed and each well was stained with Coomassie Blue to visualize and count the plaques (see above for staining).
The techniques have been described before [8], including most reagents and procedures used to stain endothelial, cells, astrocytes and microglia, as well as the methods applied in order to visualize and count endothelial tip-cell filopodia.
Because colchicine has been reported to improve the ability to visualize and count GRP-positive perikarya, which are obscured by a dense fiber plexus [9], we injected colchicine i.c.v. in rats when distributions of VIP, GRP and little SAAS would be compared directly (not Fig. 1; Fig. 2 and Table 1, only).
At the end, lungs were isolated and placed in Bouin's solution to visualize and count metastases (Leica MacroFluo Z6, Leica Microsystems, Heerbrugg, Switzerland).
Ten discontinuous sections were made for each lobe and more than 10 random fields were marked for each lobe; metastatic foci were visualized and counted using a fluorescence microscope.
Membranes were then mounted on a slide, and the nuclei of cells were stained with DAPI and visualized and counted using an inverted fluorescence microscope (Leica DMI4000B, Leica, Germany) [22].
Iba1- and Olig1-positive cells were visualized and counted as above.
Metal particles on membranes and positive immunoreactive profiles were visualized and counted manually.
