What causes autofluorescence in flow cytometry?
Cellular autofluorescence can be due to the presence of collagen and elastin, cyclic ring compounds such as NADPH and riboflavin, aromatic amino acids and cellular organelles such as mitochondria and lysosomes.
What are common sources of autofluorescence in cell media?
For starters, cell preparations themselves can be a source of endogenous autofluorescence. Some common sources are NADH, flavins, lipofuscins, collagen and elastin, as well as chlorophyll and lignin in plant samples.
How do you detect autofluorescence?
Four essential elements of fluorescence detection systems can be identified from the preceding discussion: 1) an excitation light source (Figure 5), 2) a fluorophore, 3) wavelength filters to isolate emission photons from excitation photons (Figure 5), 4) a detector that registers emission photons and produces a …
Do bacteria autofluorescence?
Among the eight bacterial strains tested, it was found that bacterial autofluorescence can vary from 80 to 1400 FITC equivalents per cell, depending on the bacterial species, and a relatively large cell-to-cell variation in autofluorescence intensity was observed.
What are the characteristics of fluorescence?
Fluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation. It is a form of luminescence. In most cases, the emitted light has a longer wavelength, and therefore a lower photon energy, than the absorbed radiation.
Why does the level of autofluorescence vary in cells?
The level in cells can vary due to variances in the levels of these cellular compounds and organelles that give rise to the fluorescence. In general, larger cells and more granular cells have increased autofluorescence due to an increase in the number of fluorescent compounds.
What is autofluorescence in AMD?
Autofluorescence is defined as the fluorescence emission observed when certain cell molecules are excited by UV or visible radiation of suitable wavelength. The FAF in AMD (FAM) Study 41 proposed a classification for FAF patterns in AMD:
What is autofluorescence in UV light?
Most plant and animal tissues show some autofluorescence when excited with ultraviolet light (e.g. light of wavelength around 365 nm). Sometimes this autofluorescence is a nuisance as it may conceal or be confused with a specific fluorescence.
What is autofluorescence in lipofuscin?
Autofluorescence is an important diagnostic characteristic of lipofuscin. Stübel (1911) was the first to observe that atrophic brown hearts fluoresced under ultraviolet light. Bommer (1929) associated fluorescence with the microscopic lipofuscin granules.