Label-free cytotoxicity screening assay by digital holographic microscopy.
We introduce a label-free technology based on digital holographic microscopy (DHM) with applicability for screening by imaging, and we demonstrate its capability for cytotoxicity assessment using mammalian living cells. For this first high content screening compatible application, we automatized a digital holographic microscope for image acquisition of cells using commercially available 96-well plates. Data generated through both label-free DHM imaging and fluorescence-based methods were in good agreement for cell viability identification and a Z'-factor close to 0.9 was determined, validating the robustness of DHM assay for phenotypic screening. Further, an excellent correlation was obtained between experimental cytotoxicity dose-response curves and known IC50 values for different toxic compounds. For comparable results, DHM has the major advantages of being label free and close to an order of magnitude faster than automated standard fluorescence microscopy.
Biomolecular Screening Facility, Swiss Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.
Jonas
J
Kühn
Etienne
E
Shaffer
Julien
J
Mena
Billy
B
Breton
Jérôme
J
Parent
Benjamin
B
Rappaz
Marc
M
Chambon
Yves
Y
Emery
Pierre
P
Magistretti
Christian
C
Depeursinge
Pierre
P
Marquet
Gerardo
G
Turcatti
D001681:Biological Assay / Q000295:instrumentation*; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006696:Holography / Q000295:instrumentation*; D007090:Image Interpretation, Computer-Assisted / Q000295:instrumentation; D021621:Imaging, Three-Dimensional / Q000295:instrumentation*; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012815:Signal Processing, Computer-Assisted / Q000295:instrumentation*; D013194:Staining and Labeling; D018675:Toxicity Tests / Q000295:instrumentation*