Fluorescence microscopy

Fluorescence microscopy

In this microscopy, microscopy is stained with fluorescent dye. Fluorescent dye is such kind of substance which absorbs light with particular wavelength and energy then emits light of different wavelength and energy i.e. lower or higher. After staining with dye then substance illuminate with blue light. The blue light is absorbed and green light emits by the dye. Exciter filter used in microscope block blue light and allow green light to pass through and reach the eye.

The fluorescent antibody technique- immunofluorescennce

The bacterial cell is incubated with specific antibody that is conjugated with a fluorescent dye; the dye antibody will cover the surface of the cell. This is a slide technique performed on slide; excess fluorescent dye antibody conjugated is washed off. This preparation is examined by ultraviolet light microscopy. Any bacterial cell which is not covered by the dye does not fluorescence and hence is not visible by this technique.

Specimen seen: Bright and colored; color of the fluorescent dye.

Commonly used for: detecting infectious agents in tissue, detecting immunological reactions.

Advantages

·         Enhances; subcellular anatomy; allows observation of motility.

·         It is more accurate and faster to examine.

Disadvantages

·         Inability to evaluate staining.

·         It is not flexible in all area, expensive and technical complexity.