The Imagestream Platform

The ImageStream platform (Amnis Corp.) is a novel flow-cytometric technology that is able to capture high-resolution images of cells in flow at rates of up to 1000 cells per second. By combining the high-throughput power of flow cytometry with its imaging capabilities, ImageStream cytometry allows for acquisition of statistically robust data on the detailed structure of cells and the location of intracellular components, effectively combining the strengths of microscopy and flow cytometry to achieve large scale detailed cell analysis.

Instruments

ImageStream-100: 405 nm, 488nm, 561 nm Excitation Lasers; 6 channel detectors; 40x objective; Extended depth of field (EDF) option ImageStream-X: 405 nm, 488 nm, 561 nm, 658 nm Excitation Lasers; 12 channel detectors; 20x, 40x, 60x objectives; Extended depth of field (EDF); autosampler (96 well plate format)

Applications

Current applications at RPCI include cell signaling studies (incl. NFκB, NFAT, STATs, beta-catenin), cell-cell interaction studies (e.g. antigen presenting cells with effector cells), intracellular trafficking of fluorescent probes or (co-)localization with cell organelles, fluorescent in situ hybridization.

Example: quantitative analysis of nuclear translocation of NFκB
The IDEAS® image analysis software applies features (algorithms) and masking
operations (region-finders), to perform image-based analysis. To assess events of
translocation, a similarity score algorithm is used. This is calculated by log
transforming the Pearsonʼs correlation coefficient for pixel values of the nuclear and
transcription factor images (eg DRAQ5 vs FITC-NFkB-p65). Changes in the
similarity score distributions are reported as an Rd metric, which is a measure for
the shift of two distributions (Fisherʼs Discriminant Ratio).
Similarity Score Rd Value

 

Imagestream

image cytometry image cytometry