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FCCS is an extension of the fluorescence correlation spectroscopy (FCS) method that uses two fluorescent molecules instead of one that emits different colours. The technique measures coincident green and red intensity fluctuations of distinct molecules that correlate if green and red labelled particles move together through a predefined confocal volume. [2]
A basic diagram of a fluorescence correlation spectroscopy instrument. The typical FCS setup consists of a laser line (wavelengths ranging typically from 405–633 nm (), and from 690–1100 nm (pulsed)), which is reflected into a microscope objective by a dichroic mirror.
In molecular spectroscopy, a Jablonski diagram is a diagram that illustrates the electronic states and often the vibrational levels of a molecule, and also the transitions between them. The states are arranged vertically by energy and grouped horizontally by spin multiplicity . [ 1 ]
A simplified Jablonski diagram illustrating the change of energy levels.. The principle behind fluorescence is that the fluorescent moiety contains electrons which can absorb a photon and briefly enter an excited state before either dispersing the energy non-radiatively or emitting it as a photon, but with a lower energy, i.e., at a longer wavelength (wavelength and energy are inversely ...
The DNA model shown (far right) is a space-filling, or CPK, model of the DNA double helix. Animated molecular models, such as the wire, or skeletal, type shown at the top of this article, allow one to visually explore the three-dimensional (3D) structure of DNA. Another type of DNA model is the space-filling, or CPK, model.
It was based on the same principle as the spectroscope, but it had a camera in place of the viewing tube. In recent years, the electronic circuits built around the photomultiplier tube have replaced the camera, allowing real-time spectrographic analysis with far greater accuracy. Arrays of photosensors are also used in place of film in ...
Selective plane illumination microscopy can also be combined with techniques such as fluorescence correlation spectroscopy, to allow spatially resolved mobility measurements of fluorescing particles (e.g. fluorescent beads, quantum dots or fluorescently labeled proteins) inside living biological samples. [20] [21]
Absorption spectrum of an aqueous solution of potassium permanganate.The spectrum consists of a series of overlapping lines belonging to a vibronic progression. Spectral line shape or spectral line profile describes the form of an electromagnetic spectrum in the vicinity of a spectral line – a region of stronger or weaker intensity in the spectrum.