1. Detection of proteins on slide preparations Histochemistry

2. Methods of detection - binding the target protein Immunological; primary antibodies secondary antibodies Immunological; primary antibodies secondary antibodies Small molecules; molecules that bind our target Small molecules; molecules that bind our target Proteins; proteins with interactions with target protein Proteins; proteins with interactions with target protein

3. Methods of visualization - marks on the target binding molecule Fluorescence; small excitable fluorescent molecules Fluorescence; small excitable fluorescent molecules Enzyme; enzymes with color reaction after adding substrate Enzyme; enzymes with color reaction after adding substrate

4. Methods of visualization - marks on the target binding molecule Fluorescence; small excitable fluorescent molecules Fluorescence; small excitable fluorescent molecules Enzyme; enzymes with color reaction after adding substrate Enzyme; enzymes with color reaction after adding substrate

5. Methods of visualization - marks on the target binding molecule Fluorescence; small excitable fluorescent molecules Fluorescence; small excitable fluorescent molecules Enzyme; enzymes with color reaction after adding substrate Enzyme; enzymes with color reaction after adding substrate eg. Alcaline phosphatase (AP) eg. Alcaline phosphatase (AP)

6. Actin - Major cytoskeletal protein responsible for movement Two forms: G-actin-one molecule F-actin-polymerized Two forms: G-actin-one molecule F-actin-polymerized Cooperation with myosin Cooperation with myosin Network through cytoplasm Network through cytoplasm Actin dynamics is necessary for life Actin dynamics is necessary for life

7. Phalloidin Poison from Amanita phalloides Poison from Amanita phalloides Specifically binds F-actin Specifically binds F-actin Can be linked with small excitable molecule without loosing ability to bind Can be linked with small excitable molecule without loosing ability to bind

8. DAPI - Fluorescent dsDNA probe Binds dsDNA in the minor groove by intercalation Binds dsDNA in the minor groove by intercalation That enhances the ability to emit light That enhances the ability to emit light Excitation UV Excitation UV Emission in blue Emission in blue

9. Double stain, actin cytoskeleton and DNA Merge from phalloidin-TRITC (red) and DAPI (blue) Merge from phalloidin-TRITC (red) and DAPI (blue)

10. Double stain, actin cytoskeleton and DNA Phalloidin-TRITC (red) and DAPI (blue)