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. 2009 Dec 1;34(23):3668-70.
doi: 10.1364/OL.34.003668.

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

YongKeun Park  1 , Toyohiko Yamauchi, Wonshik Choi, Ramachandra Dasari, Michael S Feld
Affiliations

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

YongKeun Park et al. Opt Lett. .

Abstract

We report a practical method for label-free quantification of specific molecules using spectroscopic imaging of sample-induced phase shifts. Diffraction phase microscopy equipped with various wavelengths of light source is used to record wavelength-dependent phase images. We first perform dispersion measurements on pure solutions of single molecular species present in the cells, such as albumin and hemoglobin (Hb). With this prior calibration of molecular specific dispersion, we demonstrate the extraction of Hb concentration from individual human red blood cells. The end point of this study is noninvasive monitoring of physiological states of intact living cells.

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Figures

Fig. 1
Fig. 1
(Color online) Experimental setup.
Fig. 2
Fig. 2
(Color online) (a–c) Interferograms of PDMS channel filled with Hb solution. (d–f) Corresponding quantitative phase maps (g) RIs of PDMS, BSA, and water. RIs of water are adopted from Ref. [12]. Inset, the RI difference between BSA solution and water. (h) Measured dispersion of Hb solutions (symbols) and comparison with Ref. [14] (lines). Error bars indicate standard deviations.
Fig. 3
Fig. 3
(Color online) (a–c) Quantitative phase maps of a RBC at three different wavelengths. (d) The retrieved Hb concentration. (e) Histogram of Hb concentrations and (f) cell volumes (N=25).
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