Improved phase sensitivity in spectral domain phase microscopy using line-field illumination and self phase-referencing

G. R. Harrison Spectroscopy Laboratory, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Optics Express (Impact Factor: 3.49). 06/2009; 17(13):10681-7. DOI: 10.1364/OE.17.010681
Source: PubMed


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.

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    • "Low-coherence interferometry is necessary to exclusively sample the reflection signal from the depth of interest. In the past, both spectral domain as well as time domain optical coherence tomography (OCT) based implementations of reflection phase microscopy have been reported [15–19]. Joo et al. and Choma et al. have independently developed similar setups using self phase-referenced spectral domain phase microscopy setups using point illumination [15, 16]. "
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    Optics Express 04/2011; 19(8):7587-95. DOI:10.1364/OE.19.007587 · 3.49 Impact Factor
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    • "To quantitatively obtain the phase image of the biological cells, the interferometric phase microscopy has been proposed and used in medicine and biology to observe the inner structure of the biological cells [3–7]. The interferometric phase microscopy could provide the quantitative phase images associated with the optical path delays. "
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    ABSTRACT: A slightly-off-axis interferometry based Hilbert phase microscopy (HPM) method is developed to quantitatively obtain the phase distribution. Owing to its single-shot nature and details detection ability, HPM can be used to investigate rapid phenomena that take place in transparent structures such as biological cells. Moreover, the slightly-off-axis interferometry owns higher effective bandwidth and more sensitivity than traditional off-axis interferometry. The proposed method takes advantages of the above techniques to obtain the phase image of the red blood cells and compared with the traditional off-axis interferometry and phase retrieval algorithm based on the FFT. The experimental results show that the proposed method owns fine spatial details and real-time imaging ability. We are sure that the proposed method provides a breakthrough for real-time observing and quantitative analyzing of cells in vivo.
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    ABSTRACT: Broadband interferometry is an attractive technique for the detection of cellular motions because it provides depth-resolved phase information via coherence gating. We present a phase-sensitive technique called spectral-domain phase microscopy (SDPM). SDPM is a functional extension of spectral-domain optical coherence tomography that allows for the detection of nanometer-scale motions in living cells. The sensitivity of the technique is demonstrated, and its calibration is verified. A shot-noise limit to the displacement sensitivity of this technique is derived. Measurement of cellular dynamics was performed on spontaneously beating cardiomyocytes isolated from chick embryos.
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