Digital Holographic Microscopy: A Quantitative Label-Free Microscopy Technique for Phenotypic Screening
Digital Holographic Microscopy (DHM) is a label-free imaging technique allowing visualization of transparent
cells with classical imaging cell culture plates. The quantitative DHM phase contrast image provided is related both to the
intracellular refractive index and to cell thickness.
DHM is able to distinguish cellular morphological changes on two representative cell lines (HeLa and H9c2) when treated
with doxorubicin and chloroquine, two cytotoxic compounds yielding distinct phenotypes. We analyzed parameters linked
to cell morphology and to the intracellular content in endpoint measurements and further investigated them with timelapse
recording. The results obtained by DHM were compared with other optical label-free microscopy techniques,
namely Phase Contrast, Differential Interference Contrast and Transport of Intensity Equation (reconstructed from three
bright-field images). For comparative purposes, images were acquired in a common 96-well plate format on the different
In contrast to the other microscopies assayed, images generated with DHM can be easily quantified using a simple
automatized on-the-fly analysis method for discriminating the different phenotypes generated in each cell line. The DHM
technology is suitable for the development of robust and unbiased image-based assays.
Keywords: Cell-based assay, differential interference contrast, digital holography, high-content screening, image-based screen,
phase contrast, phenotypic drug discovery.
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