Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate for point-of-care testing(POCT). Emerging microfluidics, a highly miniaturized, automatic, and int...Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate for point-of-care testing(POCT). Emerging microfluidics, a highly miniaturized, automatic, and integrated technology,are a potential substitute for traditional methods in performing rapid, low-cost, accurate, and on-site diagnoses.Molecular diagnostics are widely used in microfluidic devices as the most effective approaches for pathogen detection.This review summarizes the latest advances in microfluidics-based molecular diagnostics for infectious diseases from academic perspectives and industrial outlooks. First, we introduce the typical on-chip nucleic acid processes,including sample preprocessing, amplification, and signal read-out. Then, four categories of microfluidic platforms are compared with respect to features, merits, and demerits. We further discuss application of the digital assay in absolute nucleic acid quantification. Both the classic and recent microfluidics-based commercial molecular diagnostic devices are summarized as proof of the current market status. Finally, we propose future directions for microfluidics-based infectious disease diagnosis.展开更多
The cellular response to the complex extracellular microenvironment is highly dynamic in time and type of extracellular matrix.Accurately reconstructing this process and analyzing the changes in receptor conformation ...The cellular response to the complex extracellular microenvironment is highly dynamic in time and type of extracellular matrix.Accurately reconstructing this process and analyzing the changes in receptor conformation on the cell membrane surface and intracellular or intercellular signaling has been a major challenge in analytical chemistry and biophysical methodology.In this paper,a time-coded multiconcentration microfluidic chemical waveform generator was developed for the dynamic signaling probing with single-cell array of high temporal resolution,high throughput,and multi-concentration combination stimulation.Based on innovative microchannel structure,sophisticated external control methods and multiplexing technology,the system not only allowed for temporally sequential permutations of the four concentrations of stimuli(time code),but also generated pulsed and continuous waveforms at different frequencies in a highly controllable manner.Furthermore,the single-cell trap array was set up to efficiently capture cells in suspension,dramatically increasing throughput and reducing experiment preparation time.The maximum frequency of the platform was 1 Hz,and one cell could be stimulated at multiple frequencies.To show the ability of the system to investigate rapid biochemical events in high throughput,pulse stimulation and continuous stimulation of different frequencies and different time codes,combined with four concentrations of histamine(HA),were generated for probing G protein-coupled receptor(GPCR)signaling in He La cells.Then,statistical analysis was performed for the mean peak height and mean peak area of the cellular response.We believe that the time-coded multi-concentration microfluidic chemical waveform generator will provide a novel strategy for analytical chemistry,biophysics,cell signaling,and individualized medicine applications.展开更多
Photodynamic therapy(PDT)is a promising strategy for tumor treatment.Still,its therapeutic efficacy is compromised by the unsatisfactory cytotoxicity to specific subcellular organelles and insidious tumor microenviron...Photodynamic therapy(PDT)is a promising strategy for tumor treatment.Still,its therapeutic efficacy is compromised by the unsatisfactory cytotoxicity to specific subcellular organelles and insidious tumor microenvironment properties like hypoxia and high glutathione levels.Here,we fabricated a novel nanoenzyme that derived from metal-organic framework(MOF)with intrinsic catalase-like activities to decompose H2O2 to O2 and simultaneous glutathione consumption for enhancing PDT efficacy.The obtained Mn3O4 nanoparticle shows a larger pore size and surface area compared to native MOF particles,which can be used to load high dose photosensitizer.When decorated with AS1411 aptamer and polyethylene glycol(PEG),the obtained Mn3O4-PEG@C&A particle exhibits excellent stability and cell nucleus targeting ability.Remarkably,Mn3O4-PEG@C&A particle inhibited the tumor growth in the mouse model with high efficacy without any biotoxicity.This is the first report that applied MOF-derived nanoparticle to nucleus-targeted PDT.It may provide a new approach for designing functional nanoenzyme to subcellular organelles-targeted tumor modulation.展开更多
基金supported by the Nationa l Key Research and Development Program of China (2021YFA1101500)the National Natural Science Foundation of China (22074047)+1 种基金the Hubei Provincial Natural Science Foundation of China (2020CFB578)the Fundamental Research Funds for Central Universities,HUST (2020kfy XJJS034)。
文摘Traditional diagnostic strategies for infectious disease detection require benchtop instruments that are inappropriate for point-of-care testing(POCT). Emerging microfluidics, a highly miniaturized, automatic, and integrated technology,are a potential substitute for traditional methods in performing rapid, low-cost, accurate, and on-site diagnoses.Molecular diagnostics are widely used in microfluidic devices as the most effective approaches for pathogen detection.This review summarizes the latest advances in microfluidics-based molecular diagnostics for infectious diseases from academic perspectives and industrial outlooks. First, we introduce the typical on-chip nucleic acid processes,including sample preprocessing, amplification, and signal read-out. Then, four categories of microfluidic platforms are compared with respect to features, merits, and demerits. We further discuss application of the digital assay in absolute nucleic acid quantification. Both the classic and recent microfluidics-based commercial molecular diagnostic devices are summarized as proof of the current market status. Finally, we propose future directions for microfluidics-based infectious disease diagnosis.
基金support from the National Key Research and Development Program of China(2022YFC3400800)the National Natural Science Foundation of China(81827901)+2 种基金China Postdoctoral Science Foundation(2021M691144)Wuhan Special Preventive Medicine Research(MY19M01)Hubei Province Key Special Preventive Program(WJ2019H303).
基金the National Natural Science Foundation of China(Nos.22074047,21775049 and 31700746)the Hubei Provincial Natural Science Foundation of China(No.2020CFB578)the Fundamental Research Funds for Central Universities,HUST(Nos.2020kfy XJJS034 and 2021GCRC056)。
文摘The cellular response to the complex extracellular microenvironment is highly dynamic in time and type of extracellular matrix.Accurately reconstructing this process and analyzing the changes in receptor conformation on the cell membrane surface and intracellular or intercellular signaling has been a major challenge in analytical chemistry and biophysical methodology.In this paper,a time-coded multiconcentration microfluidic chemical waveform generator was developed for the dynamic signaling probing with single-cell array of high temporal resolution,high throughput,and multi-concentration combination stimulation.Based on innovative microchannel structure,sophisticated external control methods and multiplexing technology,the system not only allowed for temporally sequential permutations of the four concentrations of stimuli(time code),but also generated pulsed and continuous waveforms at different frequencies in a highly controllable manner.Furthermore,the single-cell trap array was set up to efficiently capture cells in suspension,dramatically increasing throughput and reducing experiment preparation time.The maximum frequency of the platform was 1 Hz,and one cell could be stimulated at multiple frequencies.To show the ability of the system to investigate rapid biochemical events in high throughput,pulse stimulation and continuous stimulation of different frequencies and different time codes,combined with four concentrations of histamine(HA),were generated for probing G protein-coupled receptor(GPCR)signaling in He La cells.Then,statistical analysis was performed for the mean peak height and mean peak area of the cellular response.We believe that the time-coded multi-concentration microfluidic chemical waveform generator will provide a novel strategy for analytical chemistry,biophysics,cell signaling,and individualized medicine applications.
基金We gratefully acknowledge the financial support from National Natural Science Foundation of China(Nos.21775049,31700746,31870856 and 31870854)National Key R&D Program of China(Nos.2017YFA0700403 and 2016YFF0100801)China Postdoctoral Science Foundation funded project(Nos.2018M630847 and 2018T110753).
文摘Photodynamic therapy(PDT)is a promising strategy for tumor treatment.Still,its therapeutic efficacy is compromised by the unsatisfactory cytotoxicity to specific subcellular organelles and insidious tumor microenvironment properties like hypoxia and high glutathione levels.Here,we fabricated a novel nanoenzyme that derived from metal-organic framework(MOF)with intrinsic catalase-like activities to decompose H2O2 to O2 and simultaneous glutathione consumption for enhancing PDT efficacy.The obtained Mn3O4 nanoparticle shows a larger pore size and surface area compared to native MOF particles,which can be used to load high dose photosensitizer.When decorated with AS1411 aptamer and polyethylene glycol(PEG),the obtained Mn3O4-PEG@C&A particle exhibits excellent stability and cell nucleus targeting ability.Remarkably,Mn3O4-PEG@C&A particle inhibited the tumor growth in the mouse model with high efficacy without any biotoxicity.This is the first report that applied MOF-derived nanoparticle to nucleus-targeted PDT.It may provide a new approach for designing functional nanoenzyme to subcellular organelles-targeted tumor modulation.
