12/8/2023 0 Comments Spectral flow cytometry![]() Where spectral flow differs most significantly from conventional flow cytometry is in the optical configuration and software analysis. ![]() Spectral detection uses gratings or prisms to disperse light across a detector array. Conventional detection uses dichroic mirrors and bandpass filters to select colors of light for detection on PMTs. Schematic comparison of conventional and spectral detection in flow cytometry. Emitted photons are then detected and analyzed by a combination of optics and software to identify unique spectral emissions from fluorophore labeled biomolecules. This allows uniform and efficient excitation by a set of monochromatic lasers. For both spectral and conventional flow cytometry methods, the process begins by delivering a sample stream through a flow chamber where cells move in a single file at a constant velocity, a process known as hydrodynamic focusing. Both systems employ standard fluidic and laser technologies that enable cell-by-cell spectral analysis. Spectral flow cytometry shares much of the same hardware associated with conventional flow cytometry. Where conventional flow cytometer instruments can detect panels with more than a dozen fluorophores, these new spectral flow cytometers are capable of accommodating multiparametric panels with upwards of 30 different fluorophores. Several companies have capitalized on spectral flow with the release of the Sony ID7000™ and the well-known Cytek® Aurora. Recent advancements in this field have led to the development of spectral flow cytometry, a rapidly growing technology with significantly enhanced multiplexing capabilities over conventional flow cytometry. At its core, flow cytometry enables researchers to obtain highly specific information on individual cells within a sample. Flow cytometry is a prolific and powerful analytical tool used for biological research and the clinic.
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