Citation: | Tian Peng, Yan Wei, Li Fanxing, et al. Biology microscopy using well-distributed sphere digital in-line holography[J]. Opto-Electronic Engineering, 2019, 46(1): 180110. doi: 10.12086/oee.2019.180110 |
[1] | 江旻珊, 张楠楠, 张学典, 等.混合搜索法在显微镜自动对焦中的应用[J].光电工程, 2017, 44(7): 685–694. doi: 10.3969/j.issn.1003-501X.2017.07.004 Jiang M S, Zhang N N, Zhang X D, et al. Applications of hybrid search strategy in microscope autofocus[J]. Opto-Electronic Engineering, 2017, 44(7): 685–694. doi: 10.3969/j.issn.1003-501X.2017.07.004 |
[2] | 芦碧波, 刘利群, 郑艳梅, 等.一种线束端子显微图像全自动分割方法[J].光电工程, 2016, 43(10): 49–55. doi: 10.3969/j.issn.1003-501X.2016.10.009 Lu B B, Liu L Q, Zheng Y M, et al. A method for segmenting the microscopic cable harness image automatically[J]. Opto-Electronic Engineering, 2016, 43(10): 49–55. doi: 10.3969/j.issn.1003-501X.2016.10.009 |
[3] | Garcia-Sucerquia J, Xu W B, Jericho S K, et al. Digital in-line holographic microscopy[J]. Applied Optics, 2006, 45(5): 836–850. doi: 10.1364/AO.45.000836 |
[4] | Xu W, Jericho M H, Meinertzhagen I A, et al. Digital in-line holography of microspheres[J]. Applied Optics, 2002, 41(25): 5367–5375. doi: 10.1364/AO.41.005367 |
[5] | Malek M, Allano D, Co tmellec S, et al. Digital in-line holography for three-dimensional-two-components particle tracking velocimetry[J]. Measurement Science and Technology, 2004, 15(4): 699–705. |
[6] | Das B, Yelleswarapu C S. Dual plane in-line digital holographic microscopy[J]. Optics Letters, 2010, 35(20): 3426–3428. doi: 10.1364/OL.35.003426 |
[7] | Zhang Y C, Xie C Q. Differential-interference-contrast digital in-line holography microscopy based on a single-optical-element[J]. Optics Letters, 2015, 40(21): 5015–5018. doi: 10.1364/OL.40.005015 |
[8] | Tian P, Hua Y L, Yang F, et al. High efficiency and flexible working distance digital in-line holographic microscopy based on Fresnel zone plate[J]. Measurement Science and Technology, 2017, 28(5): 055209. doi: 10.1088/1361-6501/aa6238 |
[9] | Kim M K. Wavelength-scanning digital interference holography for optical section imaging[J]. Optics Letters, 1999, 24(23): 1693–1695. doi: 10.1364/OL.24.001693 |
[10] | Zhang T, Yamaguchi I. Three-dimensional microscopy with phase-shifting digital holography[J]. Optics Letters, 1998, 23(15): 1221–1223. doi: 10.1364/OL.23.001221 |
[11] | Yamaguchi I, Kato J I, Ohta S, et al. Image formation in phase-shifting digital holography and applications to microscopy[J]. Applied Optics, 2001, 40(34): 6177–6186. doi: 10.1364/AO.40.006177 |
[12] | Poon T C. Recent progress in optical scanning holography[J]. Journal of Holography and Speckle, 2004, 1(1): 6–25. doi: 10.1166/jhs.2004.003 |
[13] | Yamaguchi I, Zhang T. Phase-shifting digital holography[J]. Optics Letters, 1997, 22(16): 1268–1270. doi: 10.1364/OL.22.001268 |
[14] | Das B, Yelleswarapu C S, Rao D V G L N. Quantitative phase microscopy using dual-plane in-line digital holography[J]. Applied Optics, 2012, 51(9): 1387–1395. |
[15] | Massig J H. Digital off-axis holography with a synthetic aperture[J]. Optics Letters, 2002, 27(24): 2179–2181. doi: 10.1364/OL.27.002179 |
[16] | Sánchez-Ortiga E, Doblas A, Saavedra G, et al. Off-axis digital holographic microscopy: practical design parameters for operating at diffraction limit[J]. Applied Optics, 2014, 53(10): 2058–2066. doi: 10.1364/AO.53.002058 |
Overview: Digital in-line holography (DIH) with spherical wave, originally proposed by Gabor, is the simplest way in realizing holographic. The object light and reference light are coaxial, and interference fringes pattern are recorded digitally by image sensor. Complex amplitude distribution of object are displayed through reconstruction algorithm. In visible light range, although the resolution is micron, it provides wide field of view. Moreover, the characters of fast, real-time, non-contact make it be a promising tool in material identification, biology microscopy, lab-on-a-chip applications and particle track.
The quality of point source spherical wave, emerged from pinhole, has an important impact on the imaging. However, the size and uncertain round of pinhole cannot be eliminated refer to fabrication error. Although researchers have been capable of manufacturing nanometer accuracy pinhole, the cost is expensive extremely far more than optical elements. On the other hand, wetting films, pixel super-resolution, differential-interference-contrast are applied efficiently to improve image quality, field-of-view, and resolution, but they require sophisticated operation steps far beyond the simplicity of the spherical wave digital in-line geometry.
Diffraction is much better in condition of pinhole diameter matching for incident light wavelength. And the actual size of pinhole is determined by parameters and distance of image sensor. It makes heavy demands on manufacturing accuracy. We first consider obtaining the well-distributed spherical wave. Laser focuses into an infinitesimal spot through laser beam expander and microscope objective in turn. Altering axis distance between pinhole array and microscope can obtain suitable focal spot. Matching with the pinhole, an ideal spherical wave is generated. The influence of uncertain round of pinhole can be shifted to the edge of image sensor that is negligible. Then reconstruction algorithm simplify the computational process, which presents the object information. Finally, as a proof-of-concept, biology experiments demonstrate the proposed techniques.
As shown in mosquito eggs microscopy. Figure (a) is reconstruction result without any image processing, the field of view is 3.22 mm×3.22 mm and the resolution is 5.09 μm. Figure (b) and (c) are magnified digitally four times to display single egg. The difference of reconstruction distance, the result is changing. It is called digital focusing. Besides, the whole measurement process is fairly high efficiency, because only three steps: placing object, exposure, and reconstruction. Large area and high resolution recovery image is our target, and also the characteristic of this microscopy system. It can be used in detection of micro optical element, biological recognition, path tracking of plankton etc. Especially in biology and medicine research, high efficient, flexible working distance and field of view characters are much suitable.
Optical path of well-distribute sphere
Experimental platform
5 μm horizontal and vertical lines.
Biology sample microscopy.
Mosquito eggs microscopy.