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Light sheet fluorescence microscopy (LSFM) is a fluorescence microscopy technique with an intermediate-to-high [1] optical resolution, but good optical sectioning capabilities and high speed. In contrast to epifluorescence microscopy only a thin slice (usually a few hundred nanometers to a few micrometers) of the sample is illuminated ...
[1] [2] A fluorescence microscope is 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. [3]
Multicolor fluorescence image of living HeLa cells. Fluorescence imaging is a type of non-invasive imaging technique that can help visualize biological processes taking place in a living organism. Images can be produced from a variety of methods including: microscopy, imaging probes, and spectroscopy.
Typical light sheet microscopy setup. Light sheet microscopy (LSM) was developed to allow for fine optical sectioning of thick biological samples without the need for physical sectioning or clearing, which are both time consuming and detrimental to in-vivo imaging. [10] While most fluorescent imaging techniques use aligned illumination and ...
Fluorescence-lifetime imaging microscopy or FLIM is an imaging technique based on the differences in the exponential decay rate of the photon emission of a fluorophore from a sample. It can be used as an imaging technique in confocal microscopy , two-photon excitation microscopy , and multiphoton tomography.
Green = smooth muscle actin (SMA) with Alexa 488 fluorophore. Blue = DAPI counterstain. Red = auto-fluorescence. Immunofluorescence (IF) is a light microscopy-based technique that allows detection and localization of a wide variety of target biomolecules within a cell or tissue at a quantitative level.
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 light path of a bright-field microscope is extremely simple; no additional components are required beyond the normal light-microscope setup. The light path begins at the illuminator or the light source on the base of the microscope. Often a halogen lamp is used. The light travels through the objective lens into the ocular lens, through ...