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.

[1]  Jérôme Primot,et al.  Achromatic three-wave (or more) lateral shearing interferometer , 1995 .

[2]  R. Barer.  Interference Microscopy and Mass Determination , 1952, Nature.

[3]  E. Cuche,et al.  Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms. , 1999, Applied optics.

[4]  Guillermo J. Tearney,et al.  Wavefront image sensor chip , 2010, Optics express.

[5]  A. Sergé,et al.  Dynamic multiple-target tracing to probe spatiotemporal cartography of cell membranes , 2008, Nature Methods.

[6]  Zhuo Wang,et al.  Dispersion-relation phase spectroscopy of intracellular transport , 2011, Optics express.

[7]  Daniel Carl,et al.  Parameter-optimized digital holographic microscope for high-resolution living-cell analysis. , 2004, Applied optics.

[8]  E. Cuche,et al.  Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy. , 2005, Optics express.

[9]  Zhuo Wang,et al.  Optical measurement of cycle-dependent cell growth , 2011, Proceedings of the National Academy of Sciences.

[10]  B. Wattellier,et al.  Quadriwave lateral shearing interferometry for quantitative phase microscopy of living cells. , 2009, Optics express.

[11]  Gabriel Popescu,et al.  Optical imaging of cell mass and growth dynamics. , 2008, American journal of physiology. Cell physiology.

[12]  Gabriel Popescu,et al.  Diffraction phase and fluorescence microscopy. , 2006, Optics express.

[13]  Patrik Langehanenberg,et al.  Integral refractive index determination of living suspension cells by multifocus digital holographic phase contrast microscopy. , 2007, Journal of biomedical optics.

[14]  E. Cuche,et al.  Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy. , 2005, Optics letters.

[15]  Zahid Yaqoob,et al.  Quantitative DIC microscopy using an off-axis self-interference approach. , 2010, Optics letters.

[16]  Laura Waller,et al.  Phase from chromatic aberrations. , 2010, Optics express.

[17]  Christian Depeursinge,et al.  Cell morphology and intracellular ionic homeostasis explored with a multimodal approach combining epifluorescence and digital holographic microscopy , 2010, Journal of biophotonics.

[18]  Gabriel Popescu,et al.  Fourier phase microscopy for investigation of biological structures and dynamics. , 2004, Optics letters.

[19]  Mathieu Cohen,et al.  Wave-front reconstruction from multidirectional phase derivatives generated by multilateral shearing interferometers. , 2005, Optics letters.

[20]  K. Nugent,et al.  Quantitative optical phase microscopy. , 1998, Optics letters.

[21]  Christian Depeursinge,et al.  Noninvasive characterization of the fission yeast cell cycle by monitoring dry mass with digital holographic microscopy. , 2009, Journal of biomedical optics.

[22]  J Primot,et al.  Extended hartmann test based on the pseudoguiding property of a hartmann mask completed by a phase chessboard. , 2000, Applied optics.