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.
