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Abstract
We report a quantitative phase microscope based on spectral domain optical coherence tomography and line-field illumination. The line illumination allows self phase-referencing method to reject common-mode phase noise. The quantitative phase microscope also features a separate reference arm, permitting the use of high numerical aperture (NA > 1) microscope objectives for high resolution phase measurement at multiple points along the line of illumination. We demonstrate that the path-length sensitivity of the instrument can be as good as 41 pm/square root of Hz, which makes it suitable for nanometer scale study of cell motility. We present the detection of natural motions of cell surface and two-dimensional surface profiling of a HeLa cell.
Highlights
Fast, accurate, and low noise quantitative phase microscopy is vital for the most stringent applications such as nano-scale cell membrane dynamics [1]
The reflection mode phase-sensitive methods rely on low coherence interferometry and yield phase measurement proportional to the index of refraction, n, of the sample rather than the relative index, Δn
Reflection-based phase measurement techniques promise an advantage in measurement sensitivity by a factor, 2n/Δn, over the transmission-based methods provided that the intensity of illumination source is sufficient enough to compensate for the weak signal in reflection mode
Summary
Accurate, and low noise quantitative phase microscopy is vital for the most stringent applications such as nano-scale cell membrane dynamics [1]. In the past several years, different modalities have been introduced for quantitative phase measurements [2,3,4,5,6,7,8,9,10,11] These methods can be classified into two main categories: namely, transmission and reflection mode techniques. Classical spectral-domain phase microscope (SDPM) implementations employ common path configuration in which the cell substrate, typically the glass coverslip surface farther from the biological sample, serves as a reference reflector. The common-path spectral domain phase-OCT systems, compromise the spatial resolution by using relatively low NA microscope objectives to simultaneously focus the sample and the reference reflectors. In contrast to classical SDPM, the proposed technique allows simultaneous depth-resolved phase measurement of multiple lateral points, enabling the study of spatial and temporal coherence of cell membrane motions [19] in reflection mode
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