Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics

Shaked, Natan T.; Zhu, Yizheng; Badie, Nima; Bursac, Nenad; Wax, Adam

doi:10.1117/1.3420179

Cited by 57 publications

(30 citation statements)

References 8 publications

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“…We have previously shown that WFDI is applicable for studying cancer cells, chondrocytes, neurons, and cardiomyocytes. [8][9][10][11][12][13] Mature (enucleated) red blood cells are an attractive goal for WFDI since they can be assumed to have homogeneous refractive index structure and thus RBC thickness profiles can be directly obtained from their quantitative phase profile. WFDI has been used to measure various morphological parameters of RBCs, [14][15][16][17][18] including during malaria.…”

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Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry

et al. 2011

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We have applied wide-field digital interferometry (WFDI) to examine the morphology and dynamics of live red blood cells (RBCs) from individuals who suffer from sickle cell anemia (SCA), a genetic disorder that affects the structure and mechanical properties of RBCs. WFDI is a noncontact, label-free optical microscopy approach that can yield quantitative thickness profiles of RBCs and measurements of their membrane fluctuations at the nanometer scale reflecting their stiffness. We find that RBCs from individuals with SCA are significantly stiffer than those from a healthy control. Moreover, we show that the technique is sensitive enough to distinguish classes of RBCs in SCA, including sickle RBCs with apparently normal morphology, compared to the stiffer crescent-shaped sickle RBCs. We expect that this approach will be useful for diagnosis of SCA and for determining efficacy of therapeutic agents.

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Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry

et al. 2011

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“…WFDI microscopy (also called digital holographic microscopy) has been applied to various types of biological cell systems and has recorded a diverse range of cellular phenomena [2][3][4][5][6][7][8][9][10].…”

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confidence: 99%

Quantitative analysis of three-dimensional biological cells using interferometric microscopy

Shaked

Wax

2011

SPIE Proceedings

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Live biological cells are three-dimensional microscopic objects that constantly adjust their sizes, shapes and other biophysical features. Wide-field digital interferometry (WFDI) is a holographic technique that is able to record the complex wavefront of the light which has interacted with in-vitro cells in a single camera exposure, where no exogenous contrast agents are required. However, simple quasi-three-dimensional holographic visualization of the cell phase profiles need not be the end of the process. Quantitative analysis should permit extraction of numerical parameters which are useful for cytology or medical diagnosis. Using a transmission-mode setup, the phase profile represents the multiplication between the integral refractive index and the thickness of the sample. These coupled variables may not be distinct when acquiring the phase profiles of dynamic cells. Many morphological parameters which are useful for cell biologists are based on the cell thickness profile rather than on its phase profile. We first overview methods to decouple the cell thickness and its refractive index using the WFDI-based phase profile. Then, we present a whole-cell-imaging approach which is able to extract useful numerical parameters on the cells even in cases where decoupling of cell thickness and refractive index is not possible or desired.

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“…This optical information yields biological information, including cell mass [2], [3], [4], [5], membrane fluctuations [6], [7], cell tomography [8], [9], [10], [11], intracellular transport [12], [13], tissue scattering [14], [15], [16], blood testing [17], [18], [19], cancer diagnosis [20]. Recently, a number of QPI methods have been developed for such biomedical applications [21], [22], [23], [24], [25], [26], [27], [28], [29], [30]. However, the contrast in QPI images has always been degraded by speckles resulting from using highly coherent light sources such as lasers.…”

Section: Introductionmentioning

confidence: 99%

Cardiomyocyte Imaging Using Real-Time Spatial Light Interference Microscopy (SLIM)

et al. 2013

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Spatial light interference microscopy (SLIM) is a highly sensitive quantitative phase imaging method, which is capable of unprecedented structure studies in biology and beyond. In addition to the π/2 shift introduced in phase contrast between the scattered and unscattered light from the sample, 4 phase shifts are generated in SLIM, by increments of π/2 using a reflective liquid crystal phase modulator (LCPM). As 4 phase shifted images are required to produce a quantitative phase image, the switching speed of the LCPM and the acquisition rate of the camera limit the acquisition rate and, thus, SLIM's applicability to highly dynamic samples. In this paper we present a fast SLIM setup which can image at a maximum rate of 50 frames per second and provide in real-time quantitative phase images at 50/4 = 12.5 frames per second. We use a fast LCPM for phase shifting and a fast scientific-grade complementary metal oxide semiconductor (sCMOS) camera (Andor) for imaging. We present the dispersion relation, i.e. decay rate vs. spatial mode, associated with dynamic beating cardiomyocyte cells from the quantitative phase images obtained with the real-time SLIM system.

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Reflective interferometric chamber for quantitative phase imaging of biological sample dynamics

Cited by 57 publications

References 8 publications

Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry

Quantitative microscopy and nanoscopy of sickle red blood cells performed by wide field digital interferometry

Quantitative analysis of three-dimensional biological cells using interferometric microscopy

Cardiomyocyte Imaging Using Real-Time Spatial Light Interference Microscopy (SLIM)

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