Optical detection and measurement of living cell morphometric features with single-shot quantitative phase microscopy.
We present a quadriwave lateral shearing interferometer used as a wavefront sensor and mounted on a commercial non-modified transmission white-light microscope as a quantitative phase imaging technique. The setup is designed to simultaneously make measurements with both quantitative transmission phase and fluorescence modes: phase enables enhanced contrasted visualization of the cell structure including intracellular organelles, while fluorescence allows a complete and precise identification of each component. After the characterization of the phase measurement reliability and sensitivity on calibrated samples, we use these two imaging modes to measure the characteristic optical path difference between subcellular elements (mitochondria, actin fibers, and vesicles) and cell medium, and demonstrate that phase-only information should be sufficient to identify some organelles without any labeling, like lysosomes. Proof of principle results show that the technique could be used either as a qualitative tool for the control of cells before an experiment, or for quantitative studies on morphology, behavior, and dynamics of cells or cellular components.
Aix-Marseille Université, Ecole Centrale Marseille, CNRS UMR 7249, Institut Fresnel, Marseille, France.
Pierre
P
Bon
Julien
J
Savatier
Marine
M
Merlin
Benoıt
B
Wattellier
Serge
S
Monneret
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D017076:Computer-Aided Design; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000379:methods*; D007368:Interferometry / Q000295:instrumentation*; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D008858:Microscopy, Phase-Contrast / Q000295:instrumentation*; D010865:Pilot Projects; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013347:Subcellular Fractions / Q000648:ultrastructure*