Multispectral quantitative phase imaging of human red blood cells using inexpensive narrowband multicolor LEDs.

We report multispectral phase-shifting interference microscopy for quantitative phase imaging of human red blood cells (RBCs). A wide range of wavelengths are covered by means of using multiple color light emitting diodes (LEDs) with narrow spectral bandwidth ranging from violet to deep red color. The multicolor LED light source was designed and operated sequentially, which works as a multispectral scanning light source. Corresponding to each color LED source, five phase-shifted interferograms were recorded sequentially for the measurement of phase maps, as well as the refractive index of RBCs within the entire visible region. The proposed technique provides information about the effect of wavelengths on the morphology and refractive index of human RBCs. The system does not require expensive multiple color filters or any wavelength scanning mechanism along with broadband light source.

[1]  E. Cuche,et al.  Measurement of the integral refractive index and dynamic cell morphometry of living cells with digital holographic microscopy. , 2005, Optics express.

[2]  Shovan K. Majumder,et al.  Simultaneous determination of size and refractive index of red blood cells by light scattering measurements , 2006 .

[3]  V. Tuchin,et al.  The refractive index of human hemoglobin in the visible range , 2011, Physics in medicine and biology.

[4]  Yongjin Sung,et al.  Quantitative dispersion microscopy , 2010, Biomedical optics express.

[5]  YongKeun Park,et al.  Spectroscopic phase microscopy for quantifying hemoglobin concentrations in intact red blood cells , 2009, BiOS.

[6]  Gabriel Popescu,et al.  Hilbert phase microscopy for investigating fast dynamics in transparent systems. , 2005, Optics letters.

[7]  G. Barbastathis,et al.  Transport-of-intensity approach to differential interference contrast (TI-DIC) microscopy for quantitative phase imaging. , 2010, Optics letters.

[8]  Dalip Singh Mehta,et al.  Quantitative phase imaging of human red blood cells using phase-shifting white light interference microscopy with colour fringe analysis , 2012 .

[9]  Mitsuo Takeda,et al.  Phase-crossing algorithm for white-light fringes analysis , 2006 .

[10]  Gabriel Popescu,et al.  Quantitative phase imaging of live cells using fast Fourier phase microscopy. , 2007, Applied optics.

[11]  Gabriel Popescu,et al.  Fourier phase microscopy for investigation of biological structures and dynamics. , 2004, Optics letters.

[12]  M. Daimon,et al.  Measurement of the refractive index of distilled water from the near-infrared region to the ultraviolet region. , 2007, Applied optics.

[13]  G. Pedrini,et al.  Wave-front reconstruction from a sequence of interferograms recorded at different planes. , 2005, Optics letters.

[14]  Maitreyee Roy,et al.  White-light interference microscopy: a way to obtain high lateral resolution over an extended range of heights. , 2006, Optics express.

[15]  James C. Wyant,et al.  White light interferometry , 2002, SPIE Defense + Commercial Sensing.

[16]  K. Nugent,et al.  Quantitative optical phase microscopy. , 1998, Optics letters.

[17]  YongKeun Park,et al.  Real-time quantitative phase imaging with a spatial phase-shifting algorithm. , 2011, Optics letters.

[18]  A. Asundi,et al.  Noninterferometric single-shot quantitative phase microscopy. , 2013, Optics letters.

[19]  R. Dasari,et al.  Diffraction phase microscopy for quantifying cell structure and dynamics. , 2006, Optics letters.

[20]  Gabriel Popescu,et al.  Imaging red blood cell dynamics by quantitative phase microscopy. , 2008, Blood cells, molecules & diseases.

[21]  Yizheng Zhu,et al.  Quantitative phase spectroscopy , 2012, Biomedical optics express.

[22]  S. Jacques,et al.  Measurement of single cell refractive index, dry mass, volume, and density using a transillumination microscope. , 2012, Physical review letters.

[23]  Jaeduck Jang,et al.  Dynamic spectroscopic phase microscopy for quantifying hemoglobin concentration and dynamic membrane fluctuation in red blood cells. , 2012, Optics express.