Invited Article: Digital refocusing in quantitative phase imaging for flowing red blood cells.

Quantitative phase imaging (QPI) offers high optical path length sensitivity, probing nanoscale features of live cells, but it is typically limited to imaging just few static cells at a time. To enable utility as a biomedical diagnostic modality, higher throughput is needed. To meet this need, methods for imaging cells in flow using QPI are in development. An important need for this application is to enable accurate quantitative analysis. However, this can be complicated when cells shift focal planes during flow. QPI permits digital refocusing since the complex optical field is measured. Here we analyze QPI images of moving red blood cells with an emphasis on choosing a quantitative criterion for digitally refocusing cell images. Of particular interest is the influence of optical absorption which can skew refocusing algorithms. Examples of refocusing of holographic images of flowing red blood cells using different approaches are presented and analyzed.