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FCCS measures the coincident green and red intensity fluctuations of distinct molecules that correlate if green and red labeled particles move together through a predefined confocal volume. To perform fluorescence cross-correlation spectroscopy (FCCS), samples of interest are first labeled with fluorescent probes of different colours.
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 ...
DNA Sequencing: Sanger Sequencing is a common form of nucleic acid detection that may use fluorescently labeled ddNTPs to image fluorescence peaks; Fluorescence image guided surgery: is a medical imaging approach that fluorescently labels a mass to aid in navigation. For example, indocyanine green can be used to detect lymph nodes in cancer ...
These data are formatted as FCS files, which are compatible with traditional flow cytometry software. Due to the high-dimensional nature of mass cytometry data, novel data analysis tools have been developed as well. [14] Imaging Mass Cytometry data analysis has its specificity due to different nature of data obtained. In terms of data analysis ...