Label-free imaging of intracellular motility by low-coherent quantitative phase microscope in reflection geometry

Toyohiko Yamauchi; Hidenao Iwai; Yutaka Yamashita

doi:10.1117/12.903668

28 November 2011 Label-free imaging of intracellular motility by low-coherent quantitative phase microscope in reflection geometry

Toyohiko Yamauchi, Hidenao Iwai, Yutaka Yamashita

Author Affiliations +

Proceedings Volume 8200, 2011 International Conference on Optical Instruments and Technology: Optoelectronic Imaging and Processing Technology; 820003 (2011) https://doi.org/10.1117/12.903668
Event: International Conference on Optical Instruments and Technology (OIT2011), 2011, Beijing, Beijing, China

Abstract

We demonstrate tomographic imaging of intracellular activity of living cells by a low-coherent quantitative phase microscope. The intracellular organelles, such as the nucleus, nucleolus, and mitochondria, are moving around inside living cells, driven by the cellular physiological activity. In order to visualize the intracellular motility in a label-free manner we have developed a reflection-type quantitative phase microscope which employs the phase shifting interferometric technique with a low-coherent light source. The phase shifting interferometry enables us to quantitatively measure the intensity and phase of the optical field, and the low-coherence interferometry makes it possible to selectively probe a specific sectioning plane in the cell volume. The results quantitatively revealed the depth-resolved fluctuations of intracellular surfaces so that the plasma membrane and the membranes of intracellular organelles were independently measured. The transversal and the vertical spatial resolutions were 0.56 μm and 0.93 μm, respectively, and the mechanical sensitivity of the phase measurement was 1.2 nanometers. The mean-squared displacement was applied as a statistical tool to analyze the temporal fluctuation of the intracellular organelles. To the best of our knowledge, our system visualized depth-resolved intracellular organelles motion for the first time in sub-micrometer resolution without contrast agents.

Citation Download Citation

Toyohiko Yamauchi, Hidenao Iwai, and Yutaka Yamashita "Label-free imaging of intracellular motility by low-coherent quantitative phase microscope in reflection geometry", Proc. SPIE 8200, 2011 International Conference on Optical Instruments and Technology: Optoelectronic Imaging and Processing Technology, 820003 (28 November 2011); https://doi.org/10.1117/12.903668

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