In your own words explain the basic principles of fluorescen

In your own words, explain the basic principles of fluorescence. Explain the differences between direct and indirect immunofluorescence and describe the benefits of each strategy. Also, describe one common problem that occurs while viewing a fluorescent sample.

Solution

When a substance absorbs light its energy level increases, but when it comes back to its original energy level, the excess energy is emitted in the form of light which is termed as fluorescence. On the basis of fluorophore conjugating to the primary or the secondary antibody, differentiation is made between direct and indirect immunofluorescence. One of the major problem using fluourescence is photobleaching, which causes reduction in time-span of light exposure.