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Fig. 3. PureBlu™ Nuclear Fluorescent Dye Hoechst 33342. Paraffin section of human colon adenocarcinoma (heat induced antigen retrieval with citrate buffer
(pH 6), blocked with 10% FCS) stained with Mouse Anti-Human Cytokeratin 18 Antibody (MCA1864H, green). Nuclei were counterstained with PureBlu Dye Hoechst 33342 (1351304, blue).
Fig. 4. PureBlu Dye DAPI. Paraffin section of human pancreas stained with Guinea Pig Anti-Pig Insulin Antibody (5330-0104G, red), Mouse Anti-Human Chromogranin A Antibody (MCA4773, green) and PureBlu Dye DAPI (1351303, blue).
Counterstains provide contrast as well as knowledge about localization within the tissue sample (for instance by visualizing nuclei or filamentous actin).
What to consider when selecting counterstains?
Table 4: Commonly used chromogenic counterstains.
Proteins containing cationic groups (Kim et al. 2016)
Hematoxylin (4 types - Harris’s, Mayer’s, Carazzi’s, and Gill’s)
Adapted from Kim et al. 2016 and Paul no date. Fast red/Kernechtrot has been included in the table according to IHCWorld (no date c).
Table 5: Commonly used fluorescent counterstains.