Counterstaining toluidine blue

For appropriate quantitation by microscope analysis, cells have to be counterstained (0.5% thionine acetate or 0.5% toluidine blue... 

Figure 4.3 Tissue sections, (a) A 12-week-old mouse Von Kossa-stained for mineral and counterstained with toluidine blue for tissue morphology (b) IR images of cortical bone at 16 weeks the top panel shows the integrated area under the 900-1200cm phosphate envelope. The second and third panels illustrate the mineral to matrix ratios and crystallinity, respectively. Reprinted with permission from Ref [37].

Sections developed with permanent labeling methods (e.g., with DAB) can also be counterstained using, for instance, cresyl violet or aqueous toluidine blue to stain Nissl substance. For permanent stains, the usual mounting medium is one of low viscosity so it gives a thinner adhesive layer of appropriate optical properties (e.g., Permount ). For fluorescence, a corresponding typical fluorescence-free mounting medium is preferred (e.g., VECTASHIELD ). 

Fig. 4 BrdU (a) versus pH]dT (b) in the somatosensory cortex of the macaque monkey. Each animal received a single injection of either marker at E55. In (a), BrdU is visualized with DAB (browil), and the tissue is counter-stained with thionine. In (b), pHJdT is visualized with silver, and the tissue is counterstained with aqueous toluidine blue. Both photos were taken at about the same rostrocaudal and lamina level, (c) Illustrates the distribution of labeled neurons across layers from pia top) to white matter (WM, bottom), fbe number of labeled neurons was greater in pH]dT- than in BrdU-injected animals. In addition, BrdU-labeled neurons were more widely distributed. This was consistent in different cortical areas. For further information, see [1]...

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