摘要
数字全息显微成像有别于传统光学显微成像,可根据重建全息图获取细胞的生物学参数与形貌信息,是一种有效的非接触无损三维成像技术.随着像感器的发展与硬件计算能力的提升,数字全息显微成像技术在活体生物细胞检测尤其在血红细胞检测领域取得了显著进展和突破.本文介绍了同轴、离轴以及光镊辅助离轴的数字全息显微技术,这些技术利用瑞利索末菲反向传播算法、清晰度量化算法、分水岭分割算法、数字重聚焦方法与热涨落方法等来实现血红细胞的形变、空间分布、三维体积信息的高精度提取,有助于糖尿病、心血管疾病、帕金森氏疾病等病理研究.数字全息显微成像技术实现了传统三维显微成像技术难以达到的实时性和定量化检测,由于独有的非接触、无损性特点,在细胞成像领域应用前景广阔.
Digital holographic microscopy(DHM) can obtain biological parameters and morphological information of cells by reconstructing holograms, which is different from traditional optical microscopy. The DHM is a threedimensional imaging technology which is effective, non-contact and non-destructive. With the developments of the image sensor and the computing technology, it has made significant progress in the field of living cells detection, especially for red blood cell. Compared with the technologies which are widely used in the field of cell imaging such as con-focal laser scanning microscopy, scanning near-field optical microscopy and optical coherence tomography, the DHM has the advantages including wide FOV and high-resolution to achieve higher imaging and quality. This paper introduces the principle of recording and reconstruction of digital holography,and then analyzes the performance of three reconstruction algorithms using the Fresnel method, the convolution method and the angular spectrum method. The Fresnel method is suitable for the sample size larger than the image sensor. Both the convolution method and the angular spectrum method have an optimal reconstruction distance. When the reconstruction distance is different from the optimal distance, the resolution of the reconstructed image will decrease, and the angular spectrum method is better than the convolution method in overall performance. The DHM system for RBC measurements mainly adopts the convolution algorithm or the angular spectrum algorithm to implement numerical reconstruction. The systems of the in-line DHM, the offaxis DHM and the optical tweezers combining with off-axis DHM are introduced. These techniques use algorithms including Rayleigh-Sommerfeld back-propagation, the sharpness quantification, the watershed segmentation, the numerical refocusing and the thermal fluctuation to determine the focal plane position and obtain the best reconstruction distance of the RBCs, and further detect the shape change of the RBCs and extract the information of high-resolution blood vessel shape and blood flow velocity. These techniques can even achieve the dynamic tracking and measure three-dimensional volume of RBCs in real-time which is helpful for pathological studies such as diabetes, cardiovascular disease and Parkinson’s disease. With its unique noncontact and non-destructive characteristics, the DHM realizes real-time and quantitative detection that is difficult to achieve with traditional three-dimensional microscopic imaging technologies.
作者
张益溢
吴佳琛
郝然
金尚忠
曹良才
Zhang Yi-Yi;Wu Jia-Chen;Hao Ran;Jin Shang-Zhong;Cao Liang-Cai(College of Optical and Electronic Technology,China Jiliang University,Hangzhou 310018,China;Key Laboratory of Zhejiang Province on Modern Measurement Technology and Instruments,Hangzhou 310018,China;State Key Laboratory of Precision Measurement Technology and Instruments,Tsinghua University,Beijing 100084,China)
出处
《物理学报》
SCIE
EI
CAS
CSCD
北大核心
2020年第16期77-92,共16页
Acta Physica Sinica
基金
国家重点基础研究发展计划(批准号:2018YFF0214904)
国家自然科学基金项目(批准号:61975182)
浙江省重点科技计划项目(批准号:2020C03095)
之江实验室重点研发项目(批准号:2019DE0KF01)资助的课题.
关键词
数字全息
显微成像
细胞成像
数字重聚焦
digital holography
microscope
cell imaging
digital refocusing
