One kind of novel BLMs was fabricated by patch-clamp pipette technology characterized in considerably sensitive to changes of electrochemical parameters.Detectiye currents and voltage presented linear relationship whe...One kind of novel BLMs was fabricated by patch-clamp pipette technology characterized in considerably sensitive to changes of electrochemical parameters.Detectiye currents and voltage presented linear relationship when BLMs was formed and it could be confirmed by Gramicidin method.Ion current was increased by dihexyl (C_ (12)) modified ssDNA fixed on the BLMs and also indicated linear relationship to ssDNA's concentration due to the interaction of (C_ 12)-ssDNA and BLMs.Further more,the regression equations were different from BLMs fixed with ssDNA probe and a blank control BLM in the same experimental conditions.The ssDNA probe was successfully fixed on patch-clamp pipette supported-BLMs.Based on our studies,a biosensor with reactive element of patch-clamp pipette-supported BLMs has been established.展开更多
Three-dimensional(3D)cell culture methods have been validated that can replicate the tumor environment in vivo to a large extent,providing an effective tool for studying tumors.In this study,we demonstrated the use of...Three-dimensional(3D)cell culture methods have been validated that can replicate the tumor environment in vivo to a large extent,providing an effective tool for studying tumors.In this study,we demonstrated the use of standard laboratory pipette tips as micro vessels for generating 3D cell spheroids.No microfabrication or wet-chemistry surface modifications were involved in the procedure.Spheroids consisting of single or multiple cell types were generated within 24 h just by pipetting and incubating a cell suspension in pipette tips.Scanning electron microscope and optical microscope proved that the cells grew together tightly,and suggested that while gravity force might have initiated the sedimentation of cells at the bottom of the tip,the active aggregation of cells to form tight cell-cell interactions drove the formation of spheroids.Using common laboratory micropipettes and pipette tips,the rate of spheroid generation and the generation reproducibility was characterized from five boxes each with 80 tips.The ease of transferring reagents allowed modeling of the growth of microvascular endothelial cells in tumor spheroids.Moreover,the pairing and fusion of tumor spheroids could be manipulated in the pipette tips,suggesting the potential for building and assembling heterogeneous micro-tumor tissues in vitro to mimic solid tumors in vivo.This study demonstrated that spheroids can be readily and cost-effectively generated in standard biological laboratories in a timely manner using pipette tips.展开更多
Objective:Intracerebral delivery of agents in liquid form is usually achieved through commercially available and durable metal needles.However,their size and texture may contribute to mechanical brain damage.Glass pip...Objective:Intracerebral delivery of agents in liquid form is usually achieved through commercially available and durable metal needles.However,their size and texture may contribute to mechanical brain damage.Glass pipettes with a thin tip may significantly reduce injection-associated brain damage but require access to prohibitively expensive programmable pipette pullers.This study is to remove the economic barrier to the application of minimally invasive delivery of therapeutics to the brain,such as chemical compounds,viral vectors,and cells.Methods:We took advantage of the rapid development of free educational online resources and emerging low-cost 3D printers by designing an affordable pipette puller(APP)to remove the cost obstacle.Results:We showed that our APP could produce glass pipettes with a sharp tip opening down to 20μm or less,which is sufficiently thin for the delivery of therapeutics into the brain.A pipeline from pipette pulling to brain injection using low-cost and open-source equipment was established to facilitate the application of the APP.Conclusion:In the spirit of frugal science,our device may democratize glass pipette-puling and substantially promote the application of minimally invasive and precisely controlled delivery of therapeutics to the brain for finding more effective therapies of brain diseases.展开更多
文摘One kind of novel BLMs was fabricated by patch-clamp pipette technology characterized in considerably sensitive to changes of electrochemical parameters.Detectiye currents and voltage presented linear relationship when BLMs was formed and it could be confirmed by Gramicidin method.Ion current was increased by dihexyl (C_ (12)) modified ssDNA fixed on the BLMs and also indicated linear relationship to ssDNA's concentration due to the interaction of (C_ 12)-ssDNA and BLMs.Further more,the regression equations were different from BLMs fixed with ssDNA probe and a blank control BLM in the same experimental conditions.The ssDNA probe was successfully fixed on patch-clamp pipette supported-BLMs.Based on our studies,a biosensor with reactive element of patch-clamp pipette-supported BLMs has been established.
基金supported by the National Natural Science Foundation of China(No.32171401)the Natural Science Foundation of Chongqing(No.CSTB2022NSCQ-MSX0808)the Specific Research Fund of the Innovation Platform for Academicians of Hainan Province(No.YSPTZX202126),China.
文摘Three-dimensional(3D)cell culture methods have been validated that can replicate the tumor environment in vivo to a large extent,providing an effective tool for studying tumors.In this study,we demonstrated the use of standard laboratory pipette tips as micro vessels for generating 3D cell spheroids.No microfabrication or wet-chemistry surface modifications were involved in the procedure.Spheroids consisting of single or multiple cell types were generated within 24 h just by pipetting and incubating a cell suspension in pipette tips.Scanning electron microscope and optical microscope proved that the cells grew together tightly,and suggested that while gravity force might have initiated the sedimentation of cells at the bottom of the tip,the active aggregation of cells to form tight cell-cell interactions drove the formation of spheroids.Using common laboratory micropipettes and pipette tips,the rate of spheroid generation and the generation reproducibility was characterized from five boxes each with 80 tips.The ease of transferring reagents allowed modeling of the growth of microvascular endothelial cells in tumor spheroids.Moreover,the pairing and fusion of tumor spheroids could be manipulated in the pipette tips,suggesting the potential for building and assembling heterogeneous micro-tumor tissues in vitro to mimic solid tumors in vivo.This study demonstrated that spheroids can be readily and cost-effectively generated in standard biological laboratories in a timely manner using pipette tips.
基金UMGCCC American Cancer Society Institutional Research,Grant/Award Numbers:IRG-18-160-16,NIH1R21AG077631-01,R03NS123733Maryland Stem Cell Research Fund,Grant/Award Numbers:2022-MSCRFL-5893,R03NS128459。
文摘Objective:Intracerebral delivery of agents in liquid form is usually achieved through commercially available and durable metal needles.However,their size and texture may contribute to mechanical brain damage.Glass pipettes with a thin tip may significantly reduce injection-associated brain damage but require access to prohibitively expensive programmable pipette pullers.This study is to remove the economic barrier to the application of minimally invasive delivery of therapeutics to the brain,such as chemical compounds,viral vectors,and cells.Methods:We took advantage of the rapid development of free educational online resources and emerging low-cost 3D printers by designing an affordable pipette puller(APP)to remove the cost obstacle.Results:We showed that our APP could produce glass pipettes with a sharp tip opening down to 20μm or less,which is sufficiently thin for the delivery of therapeutics into the brain.A pipeline from pipette pulling to brain injection using low-cost and open-source equipment was established to facilitate the application of the APP.Conclusion:In the spirit of frugal science,our device may democratize glass pipette-puling and substantially promote the application of minimally invasive and precisely controlled delivery of therapeutics to the brain for finding more effective therapies of brain diseases.
