This study elaborates on the design,fabrication,and data analysis details of SPEED,a recently proposed smartphonebased digital polymerase chain reaction(dPCR)device.The dPCR chips incorporate partition diameters rangi...This study elaborates on the design,fabrication,and data analysis details of SPEED,a recently proposed smartphonebased digital polymerase chain reaction(dPCR)device.The dPCR chips incorporate partition diameters ranging from 50μm to 5μm,and these partitions are organized into six distinct blocks to facilitate image processing.Due to the superior thermal conductivity of Si and its potential for mass production,the dPCR chips were fabricated on a Si substrate.A temperature control system based on a high-power density Peltier element and a preheating/cooling PCR protocol user interface shortening the thermal cycle time.The optical design employs four 470 nm light-emitting diodes as light sources,with filters and mirrors effectively managing the light emitted during PCR.An algorithm is utilized for image processing and illumination nonuniformity correction including conversion to a monochromatic format,partition identification,skew correction,and the generation of an image correction mask.We validated the device using a range of deoxyribonucleic acid targets,demonstrating its potential applicability across multiple fields.Therefore,we provide guidance and verification of the design and testing of the recently proposed SPEED device.展开更多
Heat generation or consumption is required for all biological processes.Microcalorimetry is an ultrasensitive method to measure heat change for various applications.In this paper,we aimed to review the ultrasensitive ...Heat generation or consumption is required for all biological processes.Microcalorimetry is an ultrasensitive method to measure heat change for various applications.In this paper,we aimed to review the ultrasensitive mi-crocalorimeter systems and their extensive applications in bioanalysis and energy balance monitoring.We first discussed the basic structure of microcalorimeters,including the closed system and open system,temperature sensing methods,isolation materials,and temperature stabilization.Then,we focused on their applications,such as cell metabolism research,biomolecule interaction measurement,biothermal analysis,and calorimetric detec-tion.Finally,we compared the advantages and disadvantages of commercially available microcalorimeters and their contributions to bioresearch.The development of ultrasensitive microcalorimeters provides the tools for bioanalysis at the single-cell,or even subcellular,level,as well as for precise calorimetric detection.展开更多
基金support from grant no.52150710541 provided by the National Natural Science Foundation of P.R.Chinagrant no.2018YFE0109000 provided by the Ministry of Science and Technology of the P.R.China+6 种基金H.Z.was supported by grant no.62301412 from the Natural Science Foundation of Chinagrant no.2023-JCQN-0130 from the Natural Science Basic Research Program of Shaanxi Province,P.R.Chinagrant no.2023M732815 from the P.R.China Postdoctoral Science Foundationgrant no.2023BSHEDZZ18 from the Postdoctoral Science Foundation of Shaanxi provinceM.K.was supported by grant no.LTACH19005 from the Ministry of Education,YouthSports of the Czech Republic and grant no.RVO-VFN 64165 from the Ministry of Health of the Czech RepublicMinistry of Health,Czech Republic-conceptual development of research organization 00064165,General University Hospital in Prague.
文摘This study elaborates on the design,fabrication,and data analysis details of SPEED,a recently proposed smartphonebased digital polymerase chain reaction(dPCR)device.The dPCR chips incorporate partition diameters ranging from 50μm to 5μm,and these partitions are organized into six distinct blocks to facilitate image processing.Due to the superior thermal conductivity of Si and its potential for mass production,the dPCR chips were fabricated on a Si substrate.A temperature control system based on a high-power density Peltier element and a preheating/cooling PCR protocol user interface shortening the thermal cycle time.The optical design employs four 470 nm light-emitting diodes as light sources,with filters and mirrors effectively managing the light emitted during PCR.An algorithm is utilized for image processing and illumination nonuniformity correction including conversion to a monochromatic format,partition identification,skew correction,and the generation of an image correction mask.We validated the device using a range of deoxyribonucleic acid targets,demonstrating its potential applicability across multiple fields.Therefore,we provide guidance and verification of the design and testing of the recently proposed SPEED device.
基金supported by the National Natural Science Foundation of China(52150710541)Jianguo Feng was supported by the grant number of 62201187.
文摘Heat generation or consumption is required for all biological processes.Microcalorimetry is an ultrasensitive method to measure heat change for various applications.In this paper,we aimed to review the ultrasensitive mi-crocalorimeter systems and their extensive applications in bioanalysis and energy balance monitoring.We first discussed the basic structure of microcalorimeters,including the closed system and open system,temperature sensing methods,isolation materials,and temperature stabilization.Then,we focused on their applications,such as cell metabolism research,biomolecule interaction measurement,biothermal analysis,and calorimetric detec-tion.Finally,we compared the advantages and disadvantages of commercially available microcalorimeters and their contributions to bioresearch.The development of ultrasensitive microcalorimeters provides the tools for bioanalysis at the single-cell,or even subcellular,level,as well as for precise calorimetric detection.
