Single-shot slightly-off-axis interferometry based Hilbert phase microscopy of red blood cells

A slightly-off-axis interferometry based Hilbert phase microscopy (HPM) method is developed to quantitatively obtain the phase distribution. Owing to its single-shot nature and details detection ability, HPM can be used to investigate rapid phenomena that take place in transparent structures such as biological cells. Moreover, the slightly-off-axis interferometry owns higher effective bandwidth and more sensitivity than traditional off-axis interferometry. The proposed method takes advantages of the above techniques to obtain the phase image of the red blood cells and compared with the traditional off-axis interferometry and phase retrieval algorithm based on the FFT. The experimental results show that the proposed method owns fine spatial details and real-time imaging ability. We are sure that the proposed method provides a breakthrough for real-time observing and quantitative analyzing of cells in vivo.

[1]  J. Davis,et al.  The Red Blood Cell , 1961, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[2]  E. Cuche,et al.  Spatial filtering for zero-order and twin-image elimination in digital off-axis holography. , 2000, Applied optics.

[3]  Soo-Chang Pei,et al.  The generalized radial Hilbert transform and its applications to 2D edge detection (any direction or specified directions) , 2003, 2003 IEEE International Conference on Acoustics, Speech, and Signal Processing, 2003. Proceedings. (ICASSP '03)..

[4]  Natan T Shaked,et al.  Dual-interference-channel quantitative-phase microscopy of live cell dynamics. , 2009, Optics letters.

[5]  Jong Chul Ye,et al.  Self-reference extended depth-of-field quantitative phase microscopy , 2010, BiOS.

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

[7]  G. N. Vishnyakov,et al.  Interferometric-computed microtomography of 3D phase objects , 1997, Photonics West - Biomedical Optics.

[8]  C. Fang-Yen,et al.  Optical diffraction tomography for high resolution live cell imaging. , 2009, Optics express.

[9]  C. Fang-Yen,et al.  Tomographic phase microscopy , 2008, Nature Methods.

[10]  E. Cuche,et al.  Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy. , 2005, Optics letters.

[11]  Gabriel Popescu,et al.  Tissue refractometry using Hilbert phase microscopy. , 2007, Optics letters.

[12]  E. Leith,et al.  Reconstructed Wavefronts and Communication Theory , 1962 .

[13]  Ichirou Yamaguchi,et al.  Phase-shifting digital holography , 1997 .

[14]  Keith Dillon,et al.  Depth sectioning of attenuation. , 2010, Journal of the Optical Society of America. A, Optics, image science, and vision.

[15]  Gerald A. Navratil,et al.  Nonstationary signal analysis of magnetic islands in plasmas , 1999 .

[16]  Natan T Shaked,et al.  Two-step-only phase-shifting interferometry with optimized detector bandwidth for microscopy of live cells. , 2009, Optics express.

[17]  Zahid Yaqoob,et al.  Improved phase sensitivity in spectral domain phase microscopy using line-field illumination and self phase-referencing. , 2009, Optics express.

[18]  I. Yamaguchi,et al.  Phase-shifting digital holography. , 1997, Optics letters.

[19]  C. Uzoigwe,et al.  The human erythrocyte has developed the biconcave disc shape to optimise the flow properties of the blood in the large vessels. , 2006, Medical hypotheses.

[20]  Victoria J Allan,et al.  Light Microscopy Techniques for Live Cell Imaging , 2003, Science.

[21]  L L Wheeless,et al.  Classification of red blood cells as normal, sickle, or other abnormal, using a single image analysis feature. , 1994, Cytometry.

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

[23]  H Ohzu,et al.  Hybrid holographic microscopy free of conjugate and zero-order images. , 1999, Applied optics.

[24]  Michael S Feld,et al.  Imaging voltage-dependent cell motions with heterodyne Mach-Zehnder phase microscopy. , 2007, Optics letters.

[25]  S. Peter Klinken Red Blood Cells , 2002 .

[26]  Zhuo Wang,et al.  Diffraction Phase Cytometry: blood on a CD-ROM. , 2009, Optics express.