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Label-Free, Ultrahigh-Speed, Direct Imaging and Tracking of Bionanoparticles in Live Cells by Using Coherent Brightfield Microscopy

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Label-Free Super-Resolution Microscopy

Part of the book series: Biological and Medical Physics, Biomedical Engineering ((BIOMEDICAL))

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Abstract

Many important biological phenomena, ranging from cell signaling to viral infection, are accomplished by transportation of biological substances encapsulated in native nano-sized particles. Thermal fluctuation drives nanoparticles through cellular environments; this movement is facilitated by their small size. To understand how a specific cell function can be achieved through random collisions, it is useful to know the interactions between single particles and the local environment, as determined by measuring cell dynamics at high spatial and temporal resolutions . In this chapter, a simple yet powerful wide-field optical technique, coherent brightfield (COBRI) microscopy , is presented. COBRI microscopy detects linearly scattered light from a nanoparticle through imaging-based interferometry, which enables direct observation of small biological nanoparticles in live cells without labels. Proper image post-processing further improves the detection sensitivity of small particles by removing the scattering background of cell structures. COBRI microscopy can easily operate at a high speed due to its wide-field nature and stable, indefinite scattering signal. Using COBRI, the dynamics of single virus particles and cell vesicles in live cells can be successfully captured at a microsecond temporal resolution and nanometer spatial precision in three dimensions. The ultrahigh spatiotemporal resolution and shot-noise-limited sensitivity of COBRI microscopy provide an opportunity to study the biophysics and biochemistry of live cells at the nanoscale.

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Authors and Affiliations

  1. Institute of Atomic and Molecular Sciences (IAMS), Academia Sinica, No. 1, Roosevelt Road, Section 4, Taipei, 10617, Taiwan

    Chia-Lung Hsieh

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  1. Chia-Lung Hsieh
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Correspondence to Chia-Lung Hsieh .

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  1. University of North Carolina at Charlotte, Charlotte, NC, USA

    Vasily Astratov

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Hsieh, CL. (2019). Label-Free, Ultrahigh-Speed, Direct Imaging and Tracking of Bionanoparticles in Live Cells by Using Coherent Brightfield Microscopy. In: Astratov, V. (eds) Label-Free Super-Resolution Microscopy. Biological and Medical Physics, Biomedical Engineering. Springer, Cham. https://doi.org/10.1007/978-3-030-21722-8_3

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