Imaging flow cytometry (IFC)

Imaging flow cytometry (IFC), sometimes called multispectral imaging flow cytometry (MIFC), is a combination of two techniques, flow cytometry and imaging microscopy.

Like a regular flow cytometer, IFC obtains forward- and side-scatter information and records several fluorescence signals simultaneously from micrometer-sized particulate objects in a sample stream. In IFC, however, these data are recorded by using a high-resolution microscope set-up, allowing the subsequent visualization of brightfield, darkfield and fluorescence patterns on individual or overlaid images. This technique is typically used for the analysis of whole cell populations but can also be employed to analyze protein particles or other particulate matter present in biopharmaceutical drug products.

As a rather novel technique in the field of particle characterization, IFC is mainly used as a research tool supporting more established techniques, such as flow imaging microscopy, during particle identification and trouble shooting. A clear benefit over other orthogonal techniques is the potential to connect particle morphology and fluorescence information on a particle-by-particle basis.

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D.J. Houde, A.S. Berkowitz, eds., Biophysical Characterization of Proteins in Developing Biopharmaceuticals, 1st ed., Newnes, 2014

S. Zölls, R. Tantipolphan, M. Wiggenhorn, G. Winter, W. Jiskoot, W. Friess, A. Hawe, Particles in therapeutic protein formulations, Part 1: overview of analytical methods., J. Pharm. Sci. 101 [2012] 914–35. doi:10.1002/jps.23001.