Spectroscopic phase microscopy for quantifying hemoglobin concentrations in intact red blood cells

Park, YongKeun; Yamauchi, Toyohiko; Choi, Wonshik; Dasari, Ramachandra R.; Feld, Michael S.

doi:10.1364/ol.34.003668

Cited by 184 publications

(131 citation statements)

References 19 publications

Supporting

Mentioning

122

Contrasting

Unclassified

Order By: Relevance

“…7,17,18 On the other hand, the refractive index and its spatial distribution is related to the density of cell organelles and the cytoplasm 19 and dynamic changes of intracellular solute concentrations due to osmotic stimulation 20 or growth processes, 21 and thus represents also a versatile parameter that can be utilized for label-free cell tomography. [22][23][24] Early experiments to determine the average refractive index of multiple cells were reported in 1957 by Barer et al 25 Later, confocal microscopy 26 as well as microfluidic chip based devices 27 were utilized to record data about the cellular refractive index.…”

Section: Introductionmentioning

confidence: 99%

“…In Ref. 19, the hemoglobin concentration of red blood cells was analyzed by a spectroscopic approach in which data from separate quantitative phase imaging-based refractive index measurements on known hemoglobin solutions were considered. In Ref.…”

Section: Introductionmentioning

confidence: 99%

“…18, the principle in Ref. 19 was modified for single shot measurements by using a color sensitive charge coupled device sensor. Robles et al 33 analyzed dispersion effects in red blood cells by nonlinear phase spectroscopy.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Sensing dynamic cytoplasm refractive index changes of adherent cells with quantitative phase microscopy using incorporated microspheres as optical probes

Przibilla¹,

Dartmann²,

Vollmer³

et al. 2012

J. Biomed. Opt

View full text Add to dashboard Cite

Abstract. The intracellular refractive index is an important parameter that describes the optical density of the cytoplasm and the concentration of the intracellular solutes. The refractive index of adherently grown cells is difficult to access. We present a method in which silica microspheres in living cells are used to determine the cytoplasm refractive index with quantitative phase microscopy. The reliability of our approach for refractive index retrieval is shown by data from a comparative study on osmotically stimulated adherent and suspended human pancreatic tumor cells. Results from adherent human fibro sarcoma cells demonstrate the capability of the method for sensing of dynamic refractive index changes and its usage with microfluidics.

show abstract

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Sensing dynamic cytoplasm refractive index changes of adherent cells with quantitative phase microscopy using incorporated microspheres as optical probes

Przibilla¹,

Dartmann²,

Vollmer³

et al. 2012

J. Biomed. Opt

View full text Add to dashboard Cite

show abstract

“…Each material has its own distinct RI value, and QPI can therefore provide physiological information [1] such as the structure and dynamics of cells [2,3], the quantification of specific molecules [4,5], and the dry mass [6,7]. Optical diffraction tomography (ODT) is a QPI method that enables us to visualize 3D RI distributions from multiple 2D scattered fields acquired at various illumination angles [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Learning Tomography Assessed Using Mie Theory

et al. 2018

View full text Add to dashboard Cite

In optical diffraction tomography, the multiply scattered field is a nonlinear function of the refractive index (RI) of the object. The Rytov method relies on a single-scattering propagation model and is commonly used to reconstruct images. Recently, a reconstruction model was introduced based on the beam propagation method that takes multiple scattering into account. We refer to this method as learning tomography (LT). We carry out simulations and experiments in order to assess the performance of LT over the iterative single-scattering propagation method. Each algorithm is rigorously assessed for spherical and cylinderical objects, with synthetic data generated using Mie theory. By varying the RI contrast and the size of the objects, we show that the LT reconstruction is more accurate and robust than the reconstruction based on the single-scattering propagation model. In addition, we show that LT is able to correct distortions that are evident in the Rytov-approximation-based reconstructions due to limitations in phase unwrapping. More importantly, the ability of LT to handle multiple scattering is demonstrated by simulations of multiple cylinders using Mie theory and is confirmed by experiment.

show abstract

“…For example, by introducing a DMD to the illumination part, an existing bright-field microscope can be converted into a 3-D quantitative phase microscope utilizing the quantitative phase imaging unit [34,35]. Furthermore, this method can also be combined with other QPI modalities, such as spectroscopic [36][37][38][39] and polarization-dependent [40,41] phase imaging with various incident angles because the mirror array on the DMD does not suffer from dispersion or birefringence, while liquid-crystal-based SLMs do.…”

mentioning

confidence: 99%

Active illumination using a digital micromirror device for quantitative phase imaging

et al. 2015

Self Cite

View full text Add to dashboard Cite

We present a powerful and cost-effective method for active illumination using a digital micromirror device (DMD) for quantitative phase imaging techniques. Displaying binary illumination patterns on a DMD with appropriate spatial filtering, plane waves with various illumination angles are generated and impinged onto a sample. Complex optical fields of the sample obtained with various incident angles are then measured via Mach-Zehnder interferometry, from which a high-resolution two-dimensional synthetic aperture phase image and a three-dimensional refractive index tomogram of the sample are reconstructed. We demonstrate the fast and stable illumination control capability of the proposed method by imaging colloidal spheres and biological cells, including a human red blood cell and a HeLa cell.Quantitative phase imaging (QPI) has emerged as an invaluable tool for imaging small transparent objects, such as biological cells and tissues [1,2]. QPI employs various interferometric microscopy techniques, including quantitative phase microscopy [2] and digital holographic microscopy [3], to quantitatively measure the optical phase delay of samples. In particular, the measured optical phase delay provides information about the morphological and biochemical properties of biological samples at the single-cell level. Recently, QPI techniques have been widely applied to study the pathophysiology of various biological cells and tissues, including red blood cells (RBCs) [4][5][6][7], white blood cells [8], bacteria [9][10][11], neurons [12][13][14], and cancer cells [15,16].Controlling the illumination beam is crucial in QPI. Especially for measuring 3-D refractive index (RI) tomograms [17] or 2-D highresolution synthetic aperture images [18], angles of plane wave illumination impinging onto a sample should be systematically controlled, and the corresponding light field images of the sample should be measured. Traditionally, galvanometer-based rotating mirrors have been used to control the angle of the illumination beam. A galvanometer-based rotating mirror located at the plane that is conjugate to a sample can control the angle of the incident beam by tilting the mirror with a certain electric voltage.The use of galvanometers, however, has several disadvantages. Inherently, there exists mechanical instability due to position jittering induced by electric noise and positioning error at high voltages. When a two-axis galvanometer is used, the rotational surfaces of each axis cannot be simultaneously conjugate to a sample due to its geometry, and this may induce unwanted additional quadratic phase distribution on the illumination beam that can limit the accurate measurements of 3-D RI tomograms. To solve this optical misalignment, two one-axis galvanometers can be placed at separate conjugate planes, but it requires a bulky optical setup with a long optical path, which can deteriorate phase noise. More importantly, galvanometers cannot generate acomplex wavefront; only tilting of a plane wave is permitted. Recently, a spatial lig...

show abstract

Spectroscopic phase microscopy for quantifying hemoglobin concentrations in intact red blood cells

Cited by 184 publications

References 19 publications

Sensing dynamic cytoplasm refractive index changes of adherent cells with quantitative phase microscopy using incorporated microspheres as optical probes

Sensing dynamic cytoplasm refractive index changes of adherent cells with quantitative phase microscopy using incorporated microspheres as optical probes

Learning Tomography Assessed Using Mie Theory

Active illumination using a digital micromirror device for quantitative phase imaging

Contact Info

Product

Resources

About