Kinetics and specificity of HEK293T extracellular vesicle uptake using imaging flow cytometry. Multispectral imaging flow cytometry reveals distinct frequencies of γ-H2AX foci induction in DNA double strand break repair defective human cell lines. Statistical analysis of nanoparticle dosing in a dynamic cellular system. An imaging flow cytometric method for measuring cell division history and molecular symmetry during mitosis. Cell cycle analysis using imaging flow cytometry using image properties.įilby, A. Optimization of yeast cell cycle analysis and morphological characterization by multispectral imaging flow cytometry. Noisy cell-size-correlated expression of cyclin B drives probabilistic cell-size homeostasis in fission yeast. CDK control pathways integrate cell size and ploidy information to control cell division. An imaging flow cytometry-based approach to analyse the fission yeast cell cycle in fixed cells. Imaging Flow Cytometry: Methods and Protocols (Humana Press, 2015). Discussion of masks in imaging flow cytometry.īarteneva, N. A user’s guide to multicolor flow cytometry panels for comprehensive immune profiling. Holmberg-Thyden, S., Grønbæk, K., Gang, A. Flow cytometric detection of human regulatory T cells. Compensation of imaging flow cytometry data.įazekas de St Groth, B., Zhu, E., Asad, S. Sensitivity measurement and compensation in spectral imaging. Fluorescence intensity calibration for immunophenotyping by flow cytometry. Demonstration of high gain mode in combination with imaging flow cytometry for improved EV analysis. Extended depth of field imaging for high speed cell analysis. Distinguishing modes of cell death using the ImageStream multispectral imaging flow cytometer. Digitally synthesized beat frequency multiplexing for sub-millisecond fluorescence microscopy. Virtual-freezing fluorescence imaging flow cytometry. High-throughput multiparametric imaging flow cytometry: toward diffraction-limited sub-cellular detection and monitoring of sub-cellular processes. High-throughput microfluidic imaging flow cytometry. Two-step protocol for preparing adherent cells for high-throughput flow cytometry. Quantifying nuclear p65 as a parameter for NF-κB activation: correlation between ImageStream cytometry, microscopy, and Western blot. Maguire, O., Collins, C., O’Loughlin, K., Miecznikowski, J. Cellular image analysis and imaging by flow cytometry. Beyond six colors: a new era in flow cytometry.

Flow cytometry: principles and clinical applications in hematology. Finally, we discuss the current limitations of imaging flow cytometry and the innovations and new instruments that are addressing these challenges.īrown, M. For each of these methods, we outline the processes involved in analysing typical data sets and provide details of example applications. These methods start from the use of simple single-image features and multiple channel gating strategies, followed by the design and use of custom features for phenotype classification, through to powerful machine and deep-learning methods. Focusing on the first commercially available imaging flow cytometer, the ImageStream (Luminex), we use examples from the literature to discuss the progression of the analysis methods used in imaging flow cytometry.

In this Primer, we discuss the typical imaging flow instrumentation, the form of data acquired and the typical analysis tools that can be applied to these data. The measurement of large numbers of features from the resulting images provides rich data sets that have resulted in a wide range of novel biomedical applications. Imaging flow cytometry combines the high-event-rate nature of flow cytometry with the advantages of single-cell image acquisition associated with microscopy.

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