均匀球面波数字同轴全息生物显微方法
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摘要
传统的小孔球面波数字同轴全息受小孔的不确定度影响,成像质量并不理想。本文提出一种产生均匀球面波得到宽视场高分辨的显微成像方法。激光经过扩束镜、显微物镜后聚焦成一个极小的光斑,调节针孔阵列与焦点的距离,针孔直径与焦斑相配形成理想球面波。照明被测物后,透射球面波和物体散射的物光波形成干涉条纹,由大靶面图像传感器采集。载物与不载物的图像相减去掉脏点和杂光干扰。菲涅耳逆变换重构算法恢复物体信息。生物实验证明,均匀球面波数字同轴全息能够获得高质量显微成像,视场范围3.22 mm×3.22 mm,分辨率5.09μm,其快速、非接触、灵活的放大倍率可广泛应用于光学元件检测、材料识别、生物医学领域。
Traditional pinhole spherical wave digital in-line holography has proved to be powerful imaging tools. Image quality is affected by uncertain round of pinhole. Here, we propose a well-distributed sphere wave generation method and it demonstrates wide field of view and high resolution microscopy. The laser focuses into an infinitesimal spot through laser beam expander and microscope objective. Pinhole permutation with different sizes is utilized to match the focal point, and emerges an ideal spherical wave. Interference fringes pattern, formed by reference sphere wave and scattered sphere wave of object, is collected by large area image sensor. The influence of dirty in image sensor and parasitic light is eliminated through subtraction with and without object. Fresnel inverse transformation reconstruction algorithm presents the object information. Biology microscopy experiments demonstrate that the proposed techniques increase the flexibility in producing well-distributed point light source and improve the image quality. Field of view is 3.22 mm×3.22 mm and resolution is 5.09 μm. Furthermore, adjustable field of view with magnification, fast, no-contact make it to be a promising tool in optical element measurement, material identification, biology and medicine.
Traditional pinhole spherical wave digital in-line holography has proved to be powerful imaging tools. Image quality is affected by uncertain round of pinhole. Here, we propose a well-distributed sphere wave generation method and it demonstrates wide field of view and high resolution microscopy. The laser focuses into an infinitesimal spot through laser beam expander and microscope objective. Pinhole permutation with different sizes is utilized to match the focal point, and emerges an ideal spherical wave. Interference fringes pattern, formed by reference sphere wave and scattered sphere wave of object, is collected by large area image sensor. The influence of dirty in image sensor and parasitic light is eliminated through subtraction with and without object. Fresnel inverse transformation reconstruction algorithm presents the object information. Biology microscopy experiments demonstrate that the proposed techniques increase the flexibility in producing well-distributed point light source and improve the image quality. Field of view is 3.22 mm×3.22 mm and resolution is 5.09 μm. Furthermore, adjustable field of view with magnification, fast, no-contact make it to be a promising tool in optical element measurement, material identification, biology and medicine.
引文
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[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.
[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.
[2]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.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.
[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.
[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.
[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.
[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.
[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.
[9]Kim M K.Wavelength-scanning digital interference holography for optical section imaging[J].Optics Letters,1999,24(23):1693-1695.
[10]Zhang T,Yamaguchi I.Three-dimensional microscopy with phase-shifting digital holography[J].Optics Letters,1998,23(15):1221-1223.
[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.
[12]Poon T C.Recent progress in optical scanning holography[J].Journal of Holography and Speckle,2004,1(1):6-25.
[13]Yamaguchi I,Zhang T.Phase-shifting digital holography[J].Optics Letters,1997,22(16):1268-1270.
[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.
[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.