Fluorescence litime imaging(FLIM)is an effective noninvasive bioanalytical tol based onmeasuring fuorescent lifetime of fuorophores.A growing number of FLIM studies utilizes ge-netically engineered fluorescent protein...Fluorescence litime imaging(FLIM)is an effective noninvasive bioanalytical tol based onmeasuring fuorescent lifetime of fuorophores.A growing number of FLIM studies utilizes ge-netically engineered fluorescent proteins targeted to specific subcellular structures to probe localmolecular environment,which opens new directions in cell science.This paper highlights theunconventional applications of FLIM for studies of molecular processes in diverse organelles oflive cultured cells.展开更多
Fluorescence lifetime is not only associated with the molecular structure f fuorophores,but alsostrongly depends on the environment around them,which llows fuorescence lifetime imagingmicroscopy(FLIM)to be used as a t...Fluorescence lifetime is not only associated with the molecular structure f fuorophores,but alsostrongly depends on the environment around them,which llows fuorescence lifetime imagingmicroscopy(FLIM)to be used as a tool for precise measurement of the cell or tisue microenvironment,This review introduces the basic principle of fuorescence lifetime imagingtechnology and its application in clinical medicine,including research and diagnosis of diseases inskin,brain,eyes,mouth,bone,blood vessels and cavity organs,and drug evaluation.As anoninvasive,nontoxic and nonionizing radiation technique,FLIM demonstrates excellent per-formance with high sensitivity and specificity,which allows to determine precise position of thelesion and,thus,has good potential for application in biomedical research and clinical diagnosis.展开更多
Optical imaging is a most useful and widespread technique for the investigation of the structure and function of the cellular genomes.However,an analysis of immensely convoluted and irregularly compacted DNA polymer i...Optical imaging is a most useful and widespread technique for the investigation of the structure and function of the cellular genomes.However,an analysis of immensely convoluted and irregularly compacted DNA polymer is highly challenging even by modern super-resolution microscopy approaches.Here we propose fluorescence lifetime imaging(FLIM)for the advancement of studies of genomic structure including DNA compaction,replication as well as monitoring of gene expression.The proposed FLIM assay employs two independent mechanisms for DNA compaction sensing.One mechanism relies on the inverse quadratic relation between the fluorescence lifetimes of fluorescence probes incorporated into DNA and their local refractive index,variable due to DNA compaction density.Another mechanism is based on the Förster resonance energy transfer(FRET)process between the donor and the acceptor fluorophores,both incorporated into DNA.Both these proposed mechanisms were validated in cultured cells.The obtained data unravel a significant difference in compaction of the gene-rich and gene-poor pools of genomic DNA.We show that the gene-rich DNA is loosely compacted compared to the dense DNA domains devoid of active genes.展开更多
基金supported by the National Basic Research Program of China(2015CB352005)the National Natural Science Foundation of China(61525503/61378091/61620106016)+2 种基金Guangdong Natural Science Foundation Innovation Team(2014A030312008)Hong Kong,Macao and Taiwan cooperation innovation platform and major projects of international cooperation in Colleges and Universities in Guangdong Province(2015KGJHZ002)Shenzhen Basic Research Project(JCYJ20150930104948169/JCYJ20160328144746940/GJHZ 20160226202139185).
文摘Fluorescence litime imaging(FLIM)is an effective noninvasive bioanalytical tol based onmeasuring fuorescent lifetime of fuorophores.A growing number of FLIM studies utilizes ge-netically engineered fluorescent proteins targeted to specific subcellular structures to probe localmolecular environment,which opens new directions in cell science.This paper highlights theunconventional applications of FLIM for studies of molecular processes in diverse organelles oflive cultured cells.
基金funded by the Science and Technology Planning Fundamental Research Project of Shenzhen(No.JCYJ20150324140036853)National Natural Science Foundation of China(No.61378091)+1 种基金Ningbo Natural Science Foundation Project(No.2016A610032)the Central University Basic Scientic Research Business Expenses Project(No.NSIY051405).
文摘Fluorescence lifetime is not only associated with the molecular structure f fuorophores,but alsostrongly depends on the environment around them,which llows fuorescence lifetime imagingmicroscopy(FLIM)to be used as a tool for precise measurement of the cell or tisue microenvironment,This review introduces the basic principle of fuorescence lifetime imagingtechnology and its application in clinical medicine,including research and diagnosis of diseases inskin,brain,eyes,mouth,bone,blood vessels and cavity organs,and drug evaluation.As anoninvasive,nontoxic and nonionizing radiation technique,FLIM demonstrates excellent per-formance with high sensitivity and specificity,which allows to determine precise position of thelesion and,thus,has good potential for application in biomedical research and clinical diagnosis.
基金This work has been partially supported by the National Natural Science Foundation of China(61620106016/61835009/31771584)Shenzhen International Cooperation Project(GJHZ20180928161811821/GJHZ20190822095420249).
文摘Optical imaging is a most useful and widespread technique for the investigation of the structure and function of the cellular genomes.However,an analysis of immensely convoluted and irregularly compacted DNA polymer is highly challenging even by modern super-resolution microscopy approaches.Here we propose fluorescence lifetime imaging(FLIM)for the advancement of studies of genomic structure including DNA compaction,replication as well as monitoring of gene expression.The proposed FLIM assay employs two independent mechanisms for DNA compaction sensing.One mechanism relies on the inverse quadratic relation between the fluorescence lifetimes of fluorescence probes incorporated into DNA and their local refractive index,variable due to DNA compaction density.Another mechanism is based on the Förster resonance energy transfer(FRET)process between the donor and the acceptor fluorophores,both incorporated into DNA.Both these proposed mechanisms were validated in cultured cells.The obtained data unravel a significant difference in compaction of the gene-rich and gene-poor pools of genomic DNA.We show that the gene-rich DNA is loosely compacted compared to the dense DNA domains devoid of active genes.
