Staining properties of platinum blue
Staining properties of osmium tetroxide
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Staining: Immunostaining + DAB color development only
Microscope: Light-microscopeSample: Paraffin section
Image descriptionLight-microscope image of breast cancer tissue. DAB color development only. DAB-positive areas (→: infiltrating ductal epithelial cells) appear brown.
Staining: Immunostaining + DAB color development + osmium tetroxide (OsO4) staining
Image descriptionLight-microscope image of the specimen shown in Fig. 000005a following intensification with OsO4. The image is much clearer due to the enhanced contrast between the DAB-positive and -negative areas.
Following immunostaining and DAB color development, paraffin sections of pathologic tissue samples were subjected to OsO4 intensification. As a result of this treatment, the brown color of DAB-positive areas became much darker, making the areas clearly distinct from the neighboring tissue under a light microscope. As suggested by this result, OsO4 staining also highly enhances the contrast between DAB-positive and -negative areas in LVSEM images. Thus, when OsO4-intensified tissue sections are observed at higher magnifications, the morphology of subcellular compartments, such as the nucleus, can be studied in detail. 11)
Fig. 000005a shows a light-microscope image of a specimen that was immunostained for SOD1 and developed with DAB. It is a paraffin section of pathologic tissue obtained from a patient with breast cancer (invasive ductal carcinoma). The same specimen was then treated with 2% OsO4 for 1 hour to intensify DAB staining. The result is shown in Image Fig. 000005b. The comparison of these two light-microscope images clearly indicates that OsO4 treatment enhances the contrast of DAB-positive, brown ductal epithelial cells.
11) Inaga S, et al. The 119th Annual Meeting of the Japanese Association of Anatomists (March 27 – 29, 2014: Jichi Medical University)
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