Profiling individual human red blood cells using common-path diffraction optical tomography

Abstract

Due to its strong correlation with the pathophysiology of many diseases, information about human red blood cells (RBCs) has a crucial function in hematology. Therefore, measuring and understanding the morphological, chemical, and mechanical properties of individual RBCs is a key to understanding the pathophysiology of a number of diseases in hematology, as well as to opening up new possibilities for diagnosing diseases in their early stages. In this study, we present the simultaneous and quantitative measurement of the morphological, chemical, and mechanical parameters of individual RBCs employing optical holographic microtomography. In addition, it is demonstrated that the correlation analyses of these RBC parameters provide unique information for distinguishing and understanding diseases.

Mature red blood cells: from optical model to inverse light-scattering problem

“…The extended Fung model of fifth order provides the best fits over a wide range of (V,SI,c 0 ), even for highly non-spherical particles (V = 90 fl, SI = 0.55, c 0 = 0 μm 1 ) or (V = 90 fl, SI = 0.7, c 0 = 1.2 μm 1 ), while other parametric equations do not fit well the reference profiles (data not shown). Moreover, this model accuracy is within the level of the thermal fluctuations of the RBC shape, which has been measured by common-path optical tomography to be about 40 nm [8]. To further employ the fifth-order extended Fung model we obtain coefficients C 0 -C 4 as functions of h 1 /d and h 2 /d; specifically, we set C 0 = h 1 /d (by its definition), and evaluate C 1 -C 4 by polynomial approximations: …”

“…Both approaches do not verify spherization efficiency that results in uncertainty of determined volume. A number of interference methods do allow measuring volume, shape, and hemoglobin content (refractive index) of individual RBCs [6][7][8]. However, they are based on simplifying assumptions in the light-scattering theory (thus, have hard-to-control accuracy) and are not yet ready for highthroughput applications, such as routine clinical blood analysis.…”

“…There exist a number of simple empiric parametric shape equations, which has been used in optical studies of RBCs [6,15,16]. While most of them has been to some extent verified by fitting to the experimental data, they cannot be used to reliably describe the whole range of mature RBC shapes, since the underlying data is limited to a few hundred cells [6,8]. Therefore, a physically-based model would be preferable.…”

Quantitative Phase Imaging for Label-Free Analysis of Cancer Cells—Focus on Digital Holographic Microscopy

“…Important to note is that the different setups come with slightly different benefits and limitations, making them inherently well suited to a variety of purposes: High speed acquisition for high temporal resolution of flow imaging [70], in situ imaging of growing adherent cells [21,37,56], live cell tomography [71,72], or incubator monitoring [30,73].…”

High-Resolution 3-D Refractive Index Tomography and 2-D Synthetic Aperture Imaging of Live Phytoplankton

“…(2) Even though the particular focus of this study is on static 3-D images of phytoplankton, dynamics of 3-D RI tomograms of individual phytoplankton can allow to measure intracellular dynamics of subcellular organelles [39][40][41] or dynamics fluctuation in cell membrane [42][43][44][45][46][47] which can provide abundant information about pathophysiology of phytoplankton. (3) In addition, recent advances in QPI techniques can also be further employed to investigate phytoplankton research including super-resolution imaging [48], Fourier transform light scattering technique [49][50][51][52][53], real-time visualization of 3-D RI maps [54], multi-spectral QPI [55][56][57][58][59], and polarization-sensitive QPI [60,61].…”