Author Affiliations
Abstract
1 Department of Optical Science and Engineering, Shanghai Engineering Research Center of Ultra-precision Optical Manufacturing, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), School of Information Science and Technology, Fudan University, Shanghai 200433, P. R. China
2 Department of Precision Machinery and Precision Instrumentation, University of Science and Technology of China, Hefei 230027, Anhui, P. R. China
3 Institute of Biomedical Engineering and Technology, Academy for Engineering and Technology, Fudan University, Shanghai 200433, P. R. China
4 Institute of Photonic Chips, Centre for Artificial-Intelligence Nanophotonics, School of Optical-Electrical and Computer Engineering, University of Shanghai for Science and Technology, Shanghai 200093, P. R. China
5 Ruidge Biotech Co. Ltd., No. 888, Huanhu West 2nd Road, Lin-Gang Special Area, China (Shanghai) Pilot Free Trade Zone, Shanghai 200131, P. R. China
6 Shanghai Hengxin BioTechnology, Ltd., 1688 North Guo Quan Rd, Bldg A8, Rm 801, Shanghai 200438, P. R. China
7 Centre for Artificial-Intelligence Nanophotonics, School of Optical-Electrical and Computer Engineering, University of Shanghai for Science and Technology, Shanghai 200093, P. R. China
8 Shanghai Engineering Research Center of Industrial Microorganisms, The Multiscale Research Institute of Complex Systems (MRICS), School of Life Sciences, Fudan University, Shanghai 200433, P. R. China
Microfluidic systems have been widely utilized in high-throughput biology analysis, but the difficulties in liquid manipulation and cell cultivation limit its application. This work has developed a new digital microfluidic (DMF) system for on-demand droplet control. By adopting an extending-depth-of-field (EDoF) phase modulator to the optical system, the entire depth of the microfluidic channel can be covered in one image without any refocusing process, ensuring that 95% of the particles in the droplet are captured within three shots together with shaking processes. With this system, suspension droplets are generated and droplets containing only one yeast cell can be recognized, then each single cell is cultured in the array of the chip. By observing their growth in cell numbers and the green fluorescence protein (GFP) production via fluorescence imaging, the single cell with the highest production can be identified. The results have proved the heterogeneity of yeast cells, and showed that the combined system can be applied for rapid single-cell sorting, cultivation, and analysis.
Single-cell analysis digital microfluidic (DMF) extending-depth-of-field system 
Journal of Innovative Optical Health Sciences
2023, 16(3): 2244006
张丽 1刘志佳 2费义艳 2糜岚 2,*马炯 1,2,3,**
作者单位
摘要
1 复旦大学生命科学学院复杂体系多尺度研究院,上海 200433
2 复旦大学信息科学与工程学院光科学与工程系,上海 200433
3 复旦大学工程技术研究院生物医学工程研究所,上海 200433
膨胀超分辨技术是近几年出现的一种对样品制备进行改进实现分辨率提升的超分辨技术,由于其与其他光学技术的兼容性强,可以进一步提高分辨率,引起了越来越多研究人员的关注。复合膨胀技术是膨胀超分辨技术改进的一个主要发展方向之一,膨胀结合光学波动超分辨技术(ExM‐SOFI)在复合膨胀技术中是一种受限较小且使用较为广泛的技术。为了增强现有ExM‐SOFI技术的成像效果,本课题组将成像缓冲液技术应用于ExM‐SOFI技术,以增强膨胀样品在拍摄过程中的抗淬灭能力,从而使普通染料在ExM‐SOFI中的荧光强度、荧光波动幅度和闪烁比等均有增强。微管和囊泡的染色成像结果表明,使用这种技术可以使样品在高阶SOFI中保持真实结构,伪影更少,因而高阶SOFI技术可以提升膨胀样品的最终分辨率。
生物光学 ExM‐SOFI 膨胀超分辨 光学波动超分辨 成像缓冲液 抗淬灭增强 
中国激光
2023, 50(3): 0307111
钟佳慧 1伍君鑫 2孔亚伟 2苏文华 2[ ... ]糜岚 1,2,**
作者单位
摘要
1 复旦大学工程与应用技术研究院生物医学工程技术研究所,上海 200433
2 复旦大学信息科学与工程学院光科学与工程系,上海 200433

酿酒酵母是最具有吸引力的微生物之一,监测其在不同生长时期的新陈代谢状态变化对基础生物学和工业研究都具有重要意义。依据酵母的生成曲线规律培养了不同时间的酵母细胞,基于荧光寿命显微成像(FLIM)进行自体荧光寿命图像的采集,并提出了一种基于机器学习的自动分析方法,可无标记快速鉴别年轻和衰老的酵母细胞。首先,采用深度监督U-Net实现酵母细胞的自动分割;然后,提取每个酵母细胞的荧光寿命特征和形态特征;最后,采用无监督聚类方法实现分类。实验结果表明,酵母的衰老伴随着新陈代谢的变化。FLIM作为一种无标记成像技术可应用于酵母细胞的代谢分析中,结合自动化分析流程可快速准确地区分具有不同代谢差异的细胞,为后续单细胞的筛选奠定了基础。

生物技术 荧光寿命显微成像 机器学习 图像处理 酵母细胞分类 
激光与光电子学进展
2022, 59(6): 0617019
Author Affiliations
Abstract
1 Department of Optical Science and Engineering, Shanghai Engineering Research Center of Ultra-precision Optical Manufacturing, Fudan University, Shanghai 200433, P. R. China
2 Department of Pathology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430014, P. R. China
3 State Key Laboratory of High Field Laser Physics, Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, Shanghai 201800, P. R. China
4 School of Arts and Sciences, MCPHS University, Boston, MA 02115, U. S. A.
5 Department of Gynecology, The Central Hospital of Wuhan, Tongji Medical College Huazhong University of Science and Technology, Wuhan 430014, P. R. China
The endogenous fluorophores such as reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H) and flavin adenine dinucleotide (FAD) in cells and tissues can be imaged by fluorescence lifetime imaging microscopy (FLIM) to show the tissue morphology features, as well as the biomolecular changes in microenvironment. The two important coenzymes in cellular metabolism, NAD(P)H and FAD, can be used to monitor the cellular metabolic status. This work proposed a novel method to study the uterine metabolism at the adjacent site of healthy cervix. It was found that the benign uterine tumors such as leiomyomas and adenomyosis with abnormal cell growth can be detected by measuring the fluorescence lifetime of NAD(P)H and FAD in adjacent healthy cervical tissues. This method opened a novel strategy for afflicted women to undergo the cervical biopsies instead of hysterectomies for detecting tumors, which can preserve the fertility of patients. The FLIM studying on NAD(P)H and FAD indicated the correlation between metabolism and some diseases, including diabetes, hyperthyroidism and obesity. It was also suggested that the metabolic level might be quite different for a patient with a malignant tumor history.
Autofluorescence fluorescence lifetime imaging benign uterine tumor metabolism 
Journal of Innovative Optical Health Sciences
2019, 12(5): 1940006
Author Affiliations
Abstract
1 Department of Physics, Fudan University, Shanghai 200433
2 State Key Lab for Advanced Photonic Materials and Devices, Fudan University, Shanghai 200433
3 Department of Macromolecular Science and Key Laboratory of Molecular Engineering of Polymers, Fudan University, Shanghai 200433
Hydrophilic photo luminescent semiconductor quantum dots (QDs) are novel nanometer-size probes, which may have potential using in bio-imaging for biological objects. In this work, the photo-stability of these QDs in two kinds of living cells was studied, compared with conventional biological probes such as fluorescein isothiocyanate (FITC) and green fluorescence protein (GFP). It was found that the concentration of QDs in living cells is the dominant factor for its photo-stability in biological environment. When the concentration of the intracellular QDs was high, the QDs show good photo-stability that is much better than the organic fluorescent probes. However when its concentration was low, the QDs also can be photo-bleached quickly. Thus the reaching of the certain concentration level is the critical condition for QDs in the application of bio-imaging.
110.0110 Imaging systems 170.0170 Medical optics and biotechnology 180.0180 Microscopy 
Chinese Optics Letters
2005, 3(0s): 208

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