Multi-part space-time analysis: into a single-celled "society"

(middle) discussed the synthesis of cell imaging probes with students.(left) instructs doctoral students to do experiments.1952, Wilson, an American cell biologist, suggested that "all the key questions of life have to go to the cells to find answers." More
have been closely related to cell biology since they won the Nobel Prize in Physiology or Medicine and the Chemistry Prize over the past 50 years.
as a science to study the laws of cell life activity, cell biology has been under the microscope of scientists for nearly 180 years, but cells are still a "black box" for human beings.
now, researchers are trying to clarify the "nature" of cells by studying them.
since 2014, Chinese scientists have made a series of advances on major scientific issues related to single-cell biology, supported by the National Natural Science Foundation's major project, Single-Cell Multi-Part Space-Time Analysis.if the cellular environment is compared to a society, each cell is an independent person.
in the study of human society, not only are the characteristics and behaviors of individuals worthy of attention, but also the relationships between individuals in the environment of the Institute, such as coordination or confrontational effects, and the collective behavior generated by groups, are also very important. The same is true of cellular research.
years, through cell research, scientists have developed a clear understanding of life science issues such as growth and development, genetic variation, cognition and behavior, evolution and adaptability. But, according to Lu Yuexiang, an associate professor at Tsinghua University, that is not enough.
" In previous studies, scientists have explored various symptoms of cell metabolism and life movement, such as protein expression analysis, gene transcription testing (reverse transcription PCR), etc., which are more likely to obtain an average result after observation and measurement in large samples of cells. Lu Yuexiang told China Science Daily.
, however, no two cells are exactly the same. These average results mask small differences between cells that may play a decisive role in certain key life processes such as cell differentiation and tumor development.
in order to obtain more accurate and comprehensive information about the physiological state and process of cells, researchers are looking at individual cells.
" life activities within a single cell can be considered as the result of complex chemical reactions between bioactive molecules, which determine the process of cell proliferation, differentiation, apoptosis and the occurrence, development and migration of major diseases. Lu Yuexiang analyzed.
but it is not easy to study the sophisticated interactions and regulatory networks formed by these bioactive molecules. It requires scientists to understand not only their chemical composition, but also the complex processes of their interactions, as well as the regions of influence and space-time changes at specific locations in the cellular cytors.
2014, the National Natural Science Foundation of China issued a guide to the application for a major project, "Single-cell multi-group space-time analysis", and the application of the research team organized by Zhang Xinrong, a professor in the Department of Chemistry at Tsinghua University, was approved. They condensed key scientific issues such as fluorescent probe preparation and synthesis, new space-time resolution imaging methods, and in-cell biometric interactions.
"We hope to develop new methods of fluorescence analysis suitable for space-time resolution of multiple bioactive molecules in a single cell, driving advances in life sciences and basic and clinical medical research." When it comes to scientific goals, Zhang Xinrong said.can we achieve this goal? In Zhang Xinrong's view, this needs to start from the single cell multi-group molecule space-time information acquisition method. To this end, the project team divided it into "fluorescent probe preparation and synthesis" "new space-time resolution imaging method" and "in-cell biometric interaction" three directions.
to understand the unique "society" of cells, the first thing you need is a magnifying glass that drills inside the cell to get key molecular information. Therefore, fluorescent probe preparation and synthesis is essential.
For the problem of very low-content molecular detection in single cells, Tang Bo, a professor at Shandong Normal University, constructed a number of ultra-sensitive molecular and nanofluorescent probes using techniques such as conjugate polymer signal amplification, no light source excitation, spectral red shift, nucleic acid hybrid chain amplification, etc., and realized the primary and dynamic detection of the concentration of some active molecules in cells and living organisms.
, the occurrence and development of physiological processes in cells are often not isolated events of a class of molecules, involving the participation of a variety of molecules. Therefore, the team also developed a series of two-, three- and four-part simultaneous detection of fluorescent probes, and designed multimodal probes to obtain richer imaging information.
important feature of this project is the multi-component analysis probe and imaging method constructed on the basis of frame nucleic acid by Yan Chunhai, a researcher at the Shanghai Institute of Applied Physics of the Chinese Academy of Sciences at the time. Zhang Xinrong introduced that frame nucleic acid is a kind of artificial design of structural nucleic acids, with precise size, structure, decoration and precision characteristics, through accurate chemical modification, can be a variety of small molecules and large molecule probe load to the frame nucleic acid, to achieve the controlled construction of multi-group probe.
, however, it is not easy to accurately locate and detect bioactive molecules in cells within subcellular regions.
" nuclei have a high molecular density and a particularly high background fluorescence, which can make it difficult to observe single molecules. Traditional optical microimaging resolution is not sufficient to analyze the structure of chromosome DNA. Lu Yuexiang told reporters that, especially under the premise of ultra-high spatial resolution, to achieve continuous dynamic observation, fluorescent probes and imaging methods have put forward greater challenges.
In the field of living cell ultra-resolution imaging, Sun Yujie, a researcher at Peking University's Center for Biodynatic Optical Imaging, developed a high-performance probe, Gmars-Q, to enter the dark state in light, thus extending the imaging time and an order of magnitude longer than the existing best probe for ultra-resolution imaging of living cells.
"Gmars-Q opens up design strategies for optimizing fluorescent proteins based on protein structure and dynamics. Gerd Ulrich Nienhaus, a professor at the Karlsruhe Institute of Technology in Germany, spoke highly of this.
the development of modern analytical chemistry, the application of large scientific devices has been paid more and more attention by scientists.
Relying on two synchronous radiation light sources from the Institute of High Energy Physics of the Chinese Academy of Sciences and the Shanghai Institute of Applied Physics of the Chinese Academy of Sciences, the Yan Chunhai Task Force and Gao Xueyun, a researcher at the Institute of High Energy Physics of the Chinese Academy of Sciences, carried out research on the method of imaging synchronizing radiation X-ray cells.
team built an X-ray full-field 3D imaging platform, synthesized a series of X-ray imaging probes, developed a cell imaging algorithm, and realized single-cell X-ray 3D imaging. In order to meet the challenge of single technology not being able to realize the structure and function positioning of cells at high resolution, the team also developed X-ray and ultra-resolution fluorescence joint technology, and realized the breakthrough of cell structure and functional fusion imaging under nano-resolution.
based on the development of a series of single-cell multi-part space-time analysis techniques, this project is integrated into related biological research.
have found that DNA has not only sequence information, but also three-dimensional structural information. Based on this, Xie Xiaoliang, a professor at Peking University and a foreign academician of the Chinese Academy of Sciences, has developed a new multicolor and stable marker system for chromatin DNA of living cells through the transformation of sgRNA, which enables long-term continuous observation and tracking of gene locations in living cells.
2018, this major project is a major breakthrough. Xie Xiaoliang's team published an article in Science describing their achievements in the study of the genomic structure of double mammalian cells at the single-cell level. Using the newly developed Dip-C technology, the team constructed a three-dimensional structure of the single-cell genome with high spatial resolution of human-sourced double-parent cells.
" is crucial for studying cellular function and provides research and intervention for chromosomal non-metholiosome diseases such as Down's syndrome. Xie Xiaoliang said. deep analysis of cell "society" is not only to clarify the phenomena and nature of life, scientists hope to control and use these phenomena and laws accordingly, in order to achieve the benefit of mankind. With the support of this major project, many studies have shown good prospects for social application.
and treatment of many diseases will eventually have to return to cellular levels. "In Zhang Xinrong's view, a series of single-cell multi-part space-time analysis techniques can effectively deepen people's understanding of the nature of life phenomena, but also help to understand the disease system, and thus promote the development of biomedical science and related industries.
integrated functional nanopro probe developed by the
" project provides the means and basis for the cause and diagnosis of related major diseases, which is of great significance to the early warning of diseases and the improvement of disease cure rate. Zhang Xinrong told China Science Daily that some of the created probes have been market-converted, and the fluorescence imaging technology based on the probe has become one of the key technologies for evaluating the effectiveness of Chinese medicines on major new drug creation topics in China.
For example, the "ultra-high sensitivity-reversible probe" studied by Tang Bo's team was able to trace the concentration level and dynamic change process of super-oxygen anions during the development of active levels, shortening the clinical trial cycle of the drug and improving the effectiveness of drug screening. It provides technical support for the research on the effect target and evaluation of the effect of the three new varieties of Chinese medicine, which are about to enter the clinical II. and III. stages.
And X-ray cell microimaging technology based on synchromatic radiation device, the resolution can easily reach tens of nanometers, can achieve the nano-resolution non-destructive imaging of complete cells under the large field of view, compared with fluorescence microscopy device has a huge advantage, in cell microimaging also shows great application prospects.
, however, for humans, entering a cellular "society" is a long way off. There are countless unknown mysteries waiting for scientists to explore.
Zhang Xinrong said the results of the major project provide the basis for the next step in integrating multiple analytical methods and developing high-sensitivity 3D imaging across the scale of the whole organ.
" through the development of synchronous radiation X-ray phase-electro-mirror fusion imaging, it is possible to select local feature areas under the guidance of the whole brain 3D micron accuracy map for nano-precision structural analysis, greatly reducing the blindness of high-precision neural network analysis. At specific locations, fluorescent molecular imaging and mass spectrometrical molecular analysis can also be used for further functional research. Members of the project team say Chinese scientists have been on a journey of exploration and discovery about "society".