How it Works – Multispectral Imaging

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The multispectral imaging system acquires up to 12 images per cell in three different imaging modes: brightfield, darkfield, and fluorescence. Light is collected from the cells flowing in the cuvette with custom objective lenses and relayed to the spectral decomposition element, which consists of a custom set of longpass filters in an angular array. The filters direct different spectral bands to laterally distinct channels on the CCD detector. The cell images are collected using patented time-delay integration (TDI) technology which yields fluorescence sensitivity comparable to the best flow cytometers. With this technique, an image is optically decomposed into a set of sub-images, each corresponding to a different color component and spatially isolated from the remaining sub-images.

Merck:/Freestyle/BI-Bioscience/Cell-Analysis/amnis/Amins2-images/multispectral-imaging2.jpg

Hydrodynamically focused cells are trans-illuminated by a brightfield light source and orthogonally by laser(s). A high numerical aperture (NA) objective lens collects fluorescence emissions, scattered and transmitted light from the cells. The collected light in optical space intersects with the spectral decomposition element. Light of different spectral bands leaves the decomposition element at different angles such that each band is focused onto different physical locations of the CCD camera(s). As a result, each cell image is decomposed into separate sub-images on each CCD chip based on a range of spectral wavelengths. The FlowSight collects up to 12 images on one CCD and the ImageStream®X collects up to 12 higher resolution images, six per CCD. The images are in spatial registry to enable the detailed analysis of localization and co-localization of probes on, in or between cells.

Benefits

  • A unique spectral decomposition element enables brightfield, side scatter & up to 10 fluorescent images per cell
  • Precise spatial registry produces detailed localization of signal from fluorescent probes