Fluorescence spectroscopyFluorescence spectroscopy (also known as fluorimetry or spectrofluorometry) is a type of electromagnetic spectroscopy that analyzes fluorescence from a sample. It involves using a beam of light, usually ultraviolet light, that excites the electrons in molecules of certain compounds and causes them to emit light; typically, but not necessarily, visible light. A complementary technique is absorption spectroscopy. In the special case of single molecule fluorescence spectroscopy, intensity fluctuations from the emitted light are measured from either single fluorophores, or pairs of fluorophores.
FluorescenceFluorescence 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. A perceptible example of fluorescence occurs when the absorbed radiation is in the ultraviolet region of the electromagnetic spectrum (invisible to the human eye), while the emitted light is in the visible region; this gives the fluorescent substance a distinct color that can only be seen when the substance has been exposed to UV light.
LaserA laser is a device that emits light through a process of optical amplification based on the stimulated emission of electromagnetic radiation. The word laser is an anacronym that originated as an acronym for light amplification by stimulated emission of radiation. The first laser was built in 1960 by Theodore Maiman at Hughes Research Laboratories, based on theoretical work by Charles H. Townes and Arthur Leonard Schawlow. A laser differs from other sources of light in that it emits light that is coherent.
Laser pointerA laser pointer or laser pen is a small handheld device with a power source (usually a battery) and a laser diode emitting a very narrow coherent low-powered laser beam of visible light, intended to be used to highlight something of interest by illuminating it with a small bright spot of colored light. The small width of the beam and low power of typical laser pointers make the beam itself invisible in a clean atmosphere, only showing a point of light when striking an opaque surface.
Emission spectrumThe emission spectrum of a chemical element or chemical compound is the spectrum of frequencies of electromagnetic radiation emitted due to an electron making a transition from a high energy state to a lower energy state. The photon energy of the emitted photon is equal to the energy difference between the two states. There are many possible electron transitions for each atom, and each transition has a specific energy difference. This collection of different transitions, leading to different radiated wavelengths, make up an emission spectrum.
Absorption spectroscopyAbsorption spectroscopy refers to spectroscopic techniques that measure the absorption of electromagnetic radiation, as a function of frequency or wavelength, due to its interaction with a sample. The sample absorbs energy, i.e., photons, from the radiating field. The intensity of the absorption varies as a function of frequency, and this variation is the absorption spectrum. Absorption spectroscopy is performed across the electromagnetic spectrum.
FluorometerA fluorometer, fluorimeter or fluormeter is a device used to measure parameters of visible spectrum fluorescence: its intensity and wavelength distribution of emission spectrum after excitation by a certain spectrum of light. These parameters are used to identify the presence and the amount of specific molecules in a medium. Modern fluorometers are capable of detecting fluorescent molecule concentrations as low as 1 part per trillion. Fluorescence analysis can be orders of magnitude more sensitive than other techniques.
Laser diodeA laser diode (LD, also injection laser diode or ILD, or diode laser) is a semiconductor device similar to a light-emitting diode in which a diode pumped directly with electrical current can create lasing conditions at the diode's junction. Driven by voltage, the doped p–n-transition allows for recombination of an electron with a hole. Due to the drop of the electron from a higher energy level to a lower one, radiation, in the form of an emitted photon is generated. This is spontaneous emission.
Laser pumpingLaser pumping is the act of energy transfer from an external source into the gain medium of a laser. The energy is absorbed in the medium, producing excited states in its atoms. When the number of particles in one excited state exceeds the number of particles in the ground state or a less-excited state, population inversion is achieved. In this condition, the mechanism of stimulated emission can take place and the medium can act as a laser or an optical amplifier. The pump power must be higher than the lasing threshold of the laser.
Laser medicineLaser medicine consists in the use of lasers in medical diagnosis, treatments, or therapies, such as laser photodynamic therapy, photorejuvenation, and laser surgery. The word laser stands for "light amplification by stimulated emission of radiation". The laser was invented in 1960 by Theodore Maiman, and its potential uses in medicine were subsequently explored. Lasers benefit from three interesting characteristics: directivity (multiple directional functions), impulse (possibility of operating in very short pulses) and monochromaticity.
GreenGreen is the color between cyan and yellow on the visible spectrum. It is evoked by light which has a dominant wavelength of roughly 495570 nm. In subtractive color systems, used in painting and color printing, it is created by a combination of yellow and cyan; in the RGB color model, used on television and computer screens, it is one of the additive primary colors, along with red and blue, which are mixed in different combinations to create all other colors.
X-ray fluorescenceX-ray fluorescence (XRF) is the emission of characteristic "secondary" (or fluorescent) X-rays from a material that has been excited by being bombarded with high-energy X-rays or gamma rays. The phenomenon is widely used for elemental analysis and chemical analysis, particularly in the investigation of metals, glass, ceramics and building materials, and for research in geochemistry, forensic science, archaeology and art objects such as paintings.
RedRed is the color at the long wavelength end of the visible spectrum of light, next to orange and opposite violet. It has a dominant wavelength of approximately 625–740 nanometres. It is a primary color in the RGB color model and a secondary color (made from magenta and yellow) in the CMYK color model, and is the complementary color of cyan. Reds range from the brilliant yellow-tinged scarlet and vermillion to bluish-red crimson, and vary in shade from the pale red pink to the dark red burgundy.
Laser safetyLaser radiation safety is the safe design, use and implementation of lasers to minimize the risk of laser accidents, especially those involving eye injuries. Since even relatively small amounts of laser light can lead to permanent eye injuries, the sale and usage of lasers is typically subject to government regulations. Moderate and high-power lasers are potentially hazardous because they can burn the retina, or even the skin.
Laser surgeryLaser surgery is a type of surgery that uses a laser (in contrast to using a scalpel) to cut tissue. Examples include the use of a laser scalpel in otherwise conventional surgery, and soft-tissue laser surgery, in which the laser beam vaporizes soft tissue with high water content. Laser surgery is commonly used on the eye. Techniques used include LASIK, which is used to correct near and far-sightedness in vision, and photorefractive keratectomy, a procedure which permanently reshapes the cornea using an excimer laser to remove a small amount of the human tissue.
Electromagnetic absorption by waterThe absorption of electromagnetic radiation by water depends on the state of the water. The absorption in the gas phase occurs in three regions of the spectrum. Rotational transitions are responsible for absorption in the microwave and far-infrared, vibrational transitions in the mid-infrared and near-infrared. Vibrational bands have rotational fine structure. Electronic transitions occur in the vacuum ultraviolet regions. Liquid water has no rotational spectrum but does absorb in the microwave region.
Electromagnetic spectrumThe electromagnetic spectrum is the range of frequencies (the spectrum) of electromagnetic radiation and their respective wavelengths and photon energies. The electromagnetic spectrum covers electromagnetic waves with frequencies ranging from below one hertz to above 1025 hertz, corresponding to wavelengths from thousands of kilometers down to a fraction of the size of an atomic nucleus.
Fluorescence microscopeA fluorescence microscope is an optical microscope that uses fluorescence instead of, or in addition to, scattering, reflection, and attenuation or absorption, to study the properties of organic or inorganic substances. "Fluorescence microscope" refers to any microscope that uses fluorescence to generate an image, whether it is a simple set up like an epifluorescence microscope or a more complicated design such as a confocal microscope, which uses optical sectioning to get better resolution of the fluorescence image.
Absorption (electromagnetic radiation)In physics, absorption of electromagnetic radiation is how matter (typically electrons bound in atoms) takes up a photon's energy — and so transforms electromagnetic energy into internal energy of the absorber (for example, thermal energy). A notable effect is attenuation, or the gradual reduction of the intensity of light waves as they propagate through a medium. Although the absorption of waves does not usually depend on their intensity (linear absorption), in certain conditions (optics) the medium's transparency changes by a factor that varies as a function of wave intensity, and saturable absorption (or nonlinear absorption) occurs.
Quenching (fluorescence)In chemistry, quenching refers to any process which decreases the fluorescent intensity of a given substance. A variety of processes can result in quenching, such as excited state reactions, energy transfer, complex-formation and collisions. As a consequence, quenching is often heavily dependent on pressure and temperature. Molecular oxygen, iodine ions and acrylamide are common chemical quenchers. The chloride ion is a well known quencher for quinine fluorescence.
