细胞热裂解分析的动态散斑全息三维形态实时检测方法研究

The real-time measurement of three-dimensional (3D) topography in cell lysis has important research significance and application value for systematic research of lysis mechanism. Aimed at a scientific issue on the real-time measurement importance of topography on lysis mechanism research in the cell membrane rupture analysis, the project proposed the research on real-time measurement method of 3D topography based on dynamic speckle illumination holography using double rotary diffusers, thus realizing the lysed cell’s topographical change measurement with 3D, real-time, autofocus and low speckle advantages. It main research includes the key techniques such as speckle-suppressed technique based on dynamic speckle illumination using double rotary diffusers, the multiple characteristic parameter extraction technique and analysis of 3D topographical change in cell lysis process. A measurement system of 3D topographical change of suspending cell for thermal lysis in microfluidic chip is setup. The corresponding experimental research on 3D topographical change of lysed erythrocyte is performed. Further, the 3D topographical change process of lysed erythrocyte can be obtained, which will be considered as a scientific measurement means for revealing the 3D topographical change rule of lysed erythrocyte in the thermal field. The research will provide a novel real-time measurement method of 3D topography for lysis technique in microfluidic chip, and provide theoretical evidence and scientific guidance for expounding lysis mechanism, thus exhibiting the scientific significance and practical value for promoting the deep development of lysis technique.

细胞裂解三维形态实时检测对系统研究细胞裂解机理具有重要的研究意义和应用价值。本项目围绕细胞膜破裂过程分析中的形态实时检测对裂解机理研究重要性这一科学问题，开展基于旋转双散射片动态散斑照明的全息显微三维实时检测方法研究，实现三维、实时、自聚焦、低散斑的细胞裂解形态变化检测。重点研究基于旋转双散射片动态散斑照明的散斑噪声抑制技术、细胞裂解三维形态变化的多特征参量提取技术以及形态分析等关键技术，建立一套微流控芯片悬浮状态下细胞热裂解三维形态变化测量系统，开展红细胞热裂解三维形态变化实验研究，获得细胞热裂解的三维形态变化过程，为揭示红细胞热裂解三维形态变化规律提供科学的检测手段。本项目的研究成果将为微流控芯片细胞裂解技术提供一种新的三维形态实时检测方法，为阐明细胞裂解机理提供理论依据和科学指导，从而推动裂解技术的深入发展，具有重要的科学意义和应用价值。

细胞裂解三维形态实时检测对系统研究细胞裂解机理具有重要的研究意义和应用价值。本项目提出一种基于旋转双散射片动态散斑照明的全息显微三维实时检测方法，建立了旋转双散射片散斑抑制理论模型，并对双散射片时间相关函数和空间相关函数分别进行了仿真分析，结果表明，目数越高的两个散射片，反向旋转时抑制散斑的效果越好，为实验开展提供了理论依据。设计实验系统，搭建实验平台，以微孔阵列作为样本，开展手动旋转单散射片和一动一静双散射片的散斑抑制实验研究。结果表明，手动旋转一动一静双散射片在非相关散斑图样越多的情况下，散斑抑制效果越明显，但手动旋转散射片的方法不适用于动态物体测量，限制了其应用。进而展开自动旋转单散射片和双散射片的散斑抑制实验研究，以红细胞、微孔阵列等作为样本，1500目双散射片在600rpm转速下，获得了微流控芯片上悬浮红细胞的低散斑三维成像。在实验基础上，研究相位型物体自动聚焦方法、细胞分割及参数提取方法、基于深度学习的红细胞检测方法，对多个聚集型粘连红细胞进行分类及参数提取，并开发相关软件。建立了一套三维、实时、自聚焦、低散斑的微流控芯片细胞形态检测系统，可以实现横向分辨率为亚微米级，纵向分辨率为纳米级的高精度测量，这为揭示红细胞裂解三维形态变化规律提供了科学的检测手段。本项目的研究将为微流控芯片细胞裂解技术提供一种新的三维形态实时检测方法，为阐明细胞裂解机理提供理论依据和科学指导，从而推动裂解技术的深入发展，具有重要的科学意义和应用价值。