Autofluorescence
Encyclopedia
Autofluorescence is the natural emission of light by biological entities such as mitochondria and lysosome
Lysosome
thumb|350px|Schematic of typical animal cell, showing subcellular components. [[Organelle]]s: [[nucleoli]] [[cell nucleus|nucleus]] [[ribosomes]] [[vesicle |vesicle]] rough [[endoplasmic reticulum]]...

s, and is used to distinguish the light originating from artificially added fluorescent markers (fluorophore
Fluorophore
A fluorophore, in analogy to a chromophore, is a component of a molecule which causes a molecule to be fluorescent. It is a functional group in a molecule which will absorb energy of a specific wavelength and re-emit energy at a different wavelength...

s). The most commonly observed autofluorescencing molecules are NADPH and flavin
Flavin
Flavin is the common name for a group of organic compounds based on pteridine, formed by the tricyclic heteronuclear organic ring isoalloxazine. The biochemical source is the vitamin riboflavin...

s; the extracellular matrix
Extracellular matrix
In biology, the extracellular matrix is the extracellular part of animal tissue that usually provides structural support to the animal cells in addition to performing various other important functions. The extracellular matrix is the defining feature of connective tissue in animals.Extracellular...

 can also contribute to autofluorescence due to the intrinsic properties of collagen
Collagen
Collagen is a group of naturally occurring proteins found in animals, especially in the flesh and connective tissues of mammals. It is the main component of connective tissue, and is the most abundant protein in mammals, making up about 25% to 35% of the whole-body protein content...

 and elastin
Elastin
Elastin is a protein in connective tissue that is elastic and allows many tissues in the body to resume their shape after stretching or contracting. Elastin helps skin to return to its original position when it is poked or pinched. Elastin is also an important load-bearing tissue in the bodies of...

. Generally, proteins containing an increased amount of the amino acid
Amino acid
Amino acids are molecules containing an amine group, a carboxylic acid group and a side-chain that varies between different amino acids. The key elements of an amino acid are carbon, hydrogen, oxygen, and nitrogen...

s tryptophan
Tryptophan
Tryptophan is one of the 20 standard amino acids, as well as an essential amino acid in the human diet. It is encoded in the standard genetic code as the codon UGG...

, tyrosine
Tyrosine
Tyrosine or 4-hydroxyphenylalanine, is one of the 22 amino acids that are used by cells to synthesize proteins. Its codons are UAC and UAU. It is a non-essential amino acid with a polar side group...

 and phenylalanine
Phenylalanine
Phenylalanine is an α-amino acid with the formula C6H5CH2CHCOOH. This essential amino acid is classified as nonpolar because of the hydrophobic nature of the benzyl side chain. L-Phenylalanine is an electrically neutral amino acid, one of the twenty common amino acids used to biochemically form...

 show some degree of autofluorescence.

Microscopy

Autofluorescence can be problematic in fluorescence microscopy. Light-emitting stains (such as fluorescently labelled antibodies
Antibody
An antibody, also known as an immunoglobulin, is a large Y-shaped protein used by the immune system to identify and neutralize foreign objects such as bacteria and viruses. The antibody recognizes a unique part of the foreign target, termed an antigen...

) are applied to samples
Sample (material)
In general, a sample is a limited quantity of something which is intended to be similar to and represent a larger amount of that thing. The things could be countable objects such as individual items available as units for sale, or a material not countable as individual items. Samples of countable...

 to enable visualisation of specific structures. Autofluorescence interferes with detection of specific fluorescent signals, especially when the signals of interest are very dim — it causes structures other than those of interest to become visible. In some microscopes (mainly confocal microscopes), it is possible to make use of different lifetime of the excited state
Excited state
Excitation is an elevation in energy level above an arbitrary baseline energy state. In physics there is a specific technical definition for energy level which is often associated with an atom being excited to an excited state....

s of the added fluorescent markers and the endogenous molecules to exclude most of the autofluorescence.

In a few cases, autofluorescence may actually illuminate the structures of interest, or serve as a useful diagnostic
Diagnosis
Diagnosis is the identification of the nature and cause of anything. Diagnosis is used in many different disciplines with variations in the use of logics, analytics, and experience to determine the cause and effect relationships...

 indicator. For example, cellular autofluorescence can be used as an indicator of cytotoxicity
Cytotoxicity
Cytotoxicity is the quality of being toxic to cells. Examples of toxic agents are a chemical substance, an immune cell or some types of venom .-Cell physiology:...

 without the need to add fluorescent markers. The autofluorescence of human skin
Skin
-Dermis:The dermis is the layer of skin beneath the epidermis that consists of connective tissue and cushions the body from stress and strain. The dermis is tightly connected to the epidermis by a basement membrane. It also harbors many Mechanoreceptors that provide the sense of touch and heat...

 can be used to measure the level of advanced glycation end-products (AGEs), which are present in higher quantities during several human disease
Disease
A disease is an abnormal condition affecting the body of an organism. It is often construed to be a medical condition associated with specific symptoms and signs. It may be caused by external factors, such as infectious disease, or it may be caused by internal dysfunctions, such as autoimmune...

s.

Optical imaging
Optical imaging
Optical imaging is an imaging technique.Optics usually describes the behavior of visible, ultraviolet, and infrared light used in imaging.Because light is an electromagnetic wave, similar phenomena occur in X-rays, microwaves, radio waves. Chemical imaging or molecular imaging involves inference...

 systems that utilize multispectral imaging can reduce signal degradation caused by autofluorescence while adding enhanced multiplexing
Multiplexing
The multiplexed signal is transmitted over a communication channel, which may be a physical transmission medium. The multiplexing divides the capacity of the low-level communication channel into several higher-level logical channels, one for each message signal or data stream to be transferred...

 capabilities.

Autofluorescent molecules

|style="vertical-align: bottom; width: 85px;"| Excitation
Excitation
-Science:* The excited state of an atom* The excitatory postsynaptic potential* The excitation provided with an electrical generator or alternator-Agitation:*Excitement...

 (nm)

|style="vertical-align: bottom; width: 110px;"|Fluorescence
Fluorescence
Fluorescence is the emission of light by a substance that has absorbed light or other electromagnetic radiation of a different wavelength. It is a form of luminescence. In most cases, emitted light has a longer wavelength, and therefore lower energy, than the absorbed radiation...

 (nm)

|style="vertical-align: bottom;"|Organisms

|style="vertical-align: bottom;"|Reference

|- valign="top"
| NAD(P)H
|260
|450
|All
|
|- valign="top"
|Chlorophyll
Chlorophyll
Chlorophyll is a green pigment found in almost all plants, algae, and cyanobacteria. Its name is derived from the Greek words χλωρος, chloros and φύλλον, phyllon . Chlorophyll is an extremely important biomolecule, critical in photosynthesis, which allows plants to obtain energy from light...

 
|465, 665
|673, 726
|Plants
|
|- valign="top"
|Collagen
Collagen
Collagen is a group of naturally occurring proteins found in animals, especially in the flesh and connective tissues of mammals. It is the main component of connective tissue, and is the most abundant protein in mammals, making up about 25% to 35% of the whole-body protein content...

 
|270-370
|305-450
|All
|
|- valign="top"
|Retinol
Retinol
Retinol is one of the animal forms of vitamin A. It is a diterpenoid and an alcohol. It is convertible to other forms of vitamin A, and the retinyl ester derivative of the alcohol serves as the storage form of the vitamin in animals....

 
|
|500
|Animals & bacteria
|
|- valign="top"
|Riboflavin
Riboflavin
Riboflavin, also known as vitamin B2 or additive E101, is an easily absorbed micronutrient with a key role in maintaining health in humans and animals. It is the central component of the cofactors FAD and FMN, and is therefore required by all flavoproteins. As such, vitamin B2 is required for a...

 
|
|550
|All
|
|- valign="top"
|Cholecalciferol
Cholecalciferol
Cholecalciferol is a form of vitamin D, also called vitamin D3 or calciol.It is structurally similar to steroids such as testosterone, cholesterol, and cortisol .-Forms:Vitamin D3 has several forms:...


|
|380-460
|Animals
|
|- valign="top"
|Folic acid
Folic acid
Folic acid and folate , as well as pteroyl-L-glutamic acid, pteroyl-L-glutamate, and pteroylmonoglutamic acid are forms of the water-soluble vitamin B9...


|
|450
|All
|
|- valign="top"
|Pyridoxine
Pyridoxine
Pyridoxine is one of the compounds that can be called vitamin B6, along with pyridoxal and pyridoxamine. It differs from pyridoxamine by the substituent at the '4' position. It is often used as 'pyridoxine hydrochloride'.-Chemistry:...


|
|400
|All
|
|- valign="top"
|Tyrosine
Tyrosine
Tyrosine or 4-hydroxyphenylalanine, is one of the 22 amino acids that are used by cells to synthesize proteins. Its codons are UAC and UAU. It is a non-essential amino acid with a polar side group...

 
|270
|305
|All
|
|- valign="top"
|Dityrosine
|325
|400
|Animals
|
|- valign="top"
|Excimer-like aggregate
|270
|360
|Animals
|collagen
|- valign="top"
|Glycation adduct
|370
|450
|Animals
|
|- valign="top"
|Indolamine
|
|
|Animals
|
|- valign="top"
|Lipofuscin
Lipofuscin
Lipofuscin is the name given to finely granular yellow-brown pigment granules composed of lipid-containing residues of lysosomal digestion. It is considered one of the aging or "wear-and-tear" pigments, found in the liver, kidney, heart muscle, adrenals, nerve cells, and ganglion cells...


|410-470
|500-695
|Eukaryotes
|
|- valign="top"
|Polyphenol
Polyphenol
Polyphenols are a structural class of natural, synthetic, and semisynthetic organic chemicals characterized by the presence of large multiples of phenol structural units...


|
|
|Plants
|
|- valign="top"
|Tryptophan
|280
|300-350
|All
|
|- valign="top"
|Melanin
Melanin
Melanin is a pigment that is ubiquitous in nature, being found in most organisms . In animals melanin pigments are derivatives of the amino acid tyrosine. The most common form of biological melanin is eumelanin, a brown-black polymer of dihydroxyindole carboxylic acids, and their reduced forms...


|340–400
|360–560
|Animals
|
|}
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