Abstract
We present a full-field reflection phase microscope that combines low-coherence interferometry and off-axis digital holographic microscopy (DHM). The reflection-based DHM provides highly sensitive and a single-shot imaging of cellular dynamics while the use of low coherence source provides a depth-selective measurement. The setup uniquely uses a diffraction grating in the reference arm to generate an interference image of uniform contrast over the entire field-of-view albeit low-coherence light source. We have measured the path-length sensitivity of our instrument to be approximately that makes it suitable for nanometer-scale full-field measurement of membrane dynamics in live cells.
Highlights
Bio-microrheology is the quantitative study of mechanical properties of live cells [1]
We present a full-field reflection phase microscope that combines low-coherence interferometry and off-axis digital holographic microscopy (DHM)
Another future direction includes full-field and multi-cell imaging of cellular electromotility, including cell membrane motion driven by the action potential in single mammalian cells [26]
Summary
Bio-microrheology is the quantitative study of mechanical properties of live cells [1]. A number of different techniques exist to assess membrane rheological properties of live cells These include atomic force microscopy (AFM) [5], optical and magnetic tweezers [6, 7], pipette aspiration [8,9,10], electric field deformation [11], and full-field transmission phase microscopy [12]. For most type of cells, which have complicated 3-D internal cellular structures, transmission-type optical techniques will not be suitable as they will probe a combination of membrane as well as bulk properties of cells that are difficult to decouple In this context, properly designed reflection-based phase microscopy with depth-sectioning capability can play vital role to exclusively access the membrane dynamics of nucleated cells. Reflection-based optical methods promise a 2n/Δn advantage in measurement sensitivity over the transmission-based optical techniques
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