Katarina Vulić, Giulia Amos, Tobias Ruff, Revan Kasm, Stephan J. Ihle, Joël Küchler, János Vörös and Sean Weaver
Technologies for axon guidance for in vitro disease models and bottom up investigations are increasingly being used in neuroscience research. One of the most prevalent patterning methods is using polydimethylsiloxane (PDMS) microstructures due to compatibility with microscopy and electrophysiology which enables systematic tracking of axon development with precision and efficiency. Previous investigations of these guidance platforms have noted axons tend to follow edges and avoid sharp turns; however, the specific impact of spatial constraints remains only partially explored. We investigated the influence of microchannel width beyond a constriction point, as well as the number of available microchannels, on axon growth dynamics. Further, by manipulating the size of micron/submicron-sized PDMS tunnels we investigated the space restriction that prevents growth cone penetration showing that restrictions smaller than 350 nm were sufficient to exclude axons. This research offers insights into the interplay of spatial constraints, axon development, and neural behavior. The findings are important for designing in vitro platforms and in vivo neural interfaces for both fundamental neuroscience and translational applications in rapidly evolving neural implant technologies.
{"title":"Impact of microchannel width on axons for brain-on-chip applications†","authors":"Katarina Vulić, Giulia Amos, Tobias Ruff, Revan Kasm, Stephan J. Ihle, Joël Küchler, János Vörös and Sean Weaver","doi":"10.1039/D4LC00440J","DOIUrl":"10.1039/D4LC00440J","url":null,"abstract":"<p >Technologies for axon guidance for <em>in vitro</em> disease models and bottom up investigations are increasingly being used in neuroscience research. One of the most prevalent patterning methods is using polydimethylsiloxane (PDMS) microstructures due to compatibility with microscopy and electrophysiology which enables systematic tracking of axon development with precision and efficiency. Previous investigations of these guidance platforms have noted axons tend to follow edges and avoid sharp turns; however, the specific impact of spatial constraints remains only partially explored. We investigated the influence of microchannel width beyond a constriction point, as well as the number of available microchannels, on axon growth dynamics. Further, by manipulating the size of micron/submicron-sized PDMS tunnels we investigated the space restriction that prevents growth cone penetration showing that restrictions smaller than 350 nm were sufficient to exclude axons. This research offers insights into the interplay of spatial constraints, axon development, and neural behavior. The findings are important for designing <em>in vitro</em> platforms and <em>in vivo</em> neural interfaces for both fundamental neuroscience and translational applications in rapidly evolving neural implant technologies.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 22","pages":" 5155-5166"},"PeriodicalIF":6.1,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d4lc00440j?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142487387","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Felix Ansah, Marziyeh Hajialyani, Fatemeh Ahmadi, Yuming Gu, Ergun Alperay Tarım, Michael G Mauk, Gordon Awandare, Haim H Bau
Effective diagnosis of comorbidities and infectious diseases that present similar symptoms requires point-of-need assays capable of co-detecting and differentiating among multiple co-endemic pathogens to enable timely, precision medicine and effective control measures. We previously developed a two-stage isothermal amplification assay dubbed Penn-RAMP to address this need. Penn-RAMP’s first stage comprises a Recombinase Polymerase Amplification (RPA), which amplifies all targets of interest in a single reaction chamber for a short duration. The RPA amplicons are then aliquoted into multiple Loop-Mediated Isothermal Amplification (LAMP) reaction chambers, each customized with pre-dried primers to amplify a single target or a group of targets. To enable Penn-RAMP at the point of need, we describe here a self-actuated Penn-RAMP chiplet that accommodates the Penn-RAMP assay. Our chiplet employs temperature-controlled phase change valves and capillary valves to self-aliquot first-stage amplicons into multiple (five) second-stage reaction chambers and to seal these chambers. The functionality of our device is demonstrated by co-detecting plant pathogens. The analytical performance of our chiplet is comparable to that of the benchtop Penn-RAMP assay and surpasses that of standalone LAMP assays. Our self-actuated chiplet can be operated standalone with purified nucleic acids or as the downstream amplification module of a sample preparation cassette.
{"title":"Self-actuated microfluidic chiplet for two-stage multiplex nucleic acid amplification assay","authors":"Felix Ansah, Marziyeh Hajialyani, Fatemeh Ahmadi, Yuming Gu, Ergun Alperay Tarım, Michael G Mauk, Gordon Awandare, Haim H Bau","doi":"10.1039/d4lc00752b","DOIUrl":"https://doi.org/10.1039/d4lc00752b","url":null,"abstract":"Effective diagnosis of comorbidities and infectious diseases that present similar symptoms requires point-of-need assays capable of co-detecting and differentiating among multiple co-endemic pathogens to enable timely, precision medicine and effective control measures. We previously developed a two-stage isothermal amplification assay dubbed Penn-RAMP to address this need. Penn-RAMP’s first stage comprises a Recombinase Polymerase Amplification (RPA), which amplifies all targets of interest in a single reaction chamber for a short duration. The RPA amplicons are then aliquoted into multiple Loop-Mediated Isothermal Amplification (LAMP) reaction chambers, each customized with pre-dried primers to amplify a single target or a group of targets. To enable Penn-RAMP at the point of need, we describe here a self-actuated Penn-RAMP chiplet that accommodates the Penn-RAMP assay. Our chiplet employs temperature-controlled phase change valves and capillary valves to self-aliquot first-stage amplicons into multiple (five) second-stage reaction chambers and to seal these chambers. The functionality of our device is demonstrated by co-detecting plant pathogens. The analytical performance of our chiplet is comparable to that of the benchtop Penn-RAMP assay and surpasses that of standalone LAMP assays. Our self-actuated chiplet can be operated standalone with purified nucleic acids or as the downstream amplification module of a sample preparation cassette.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"82 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-10-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142486974","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The spatial organization of biophysical and biochemical cues in the extracellular matrix (ECM) in concert with reciprocal cell-cell signaling is vital to tissue patterning during development. However, elucidating the role an individual microenvironmental factor plays using existing textit{in vivo} models is difficult due to their inherent complexity. In this work, we have developed a microphysiological system to spatially pattern the biochemical, biophysical, and stromal cell composition of the ECM along an epithelialized 3D microchannel. This technique is adaptable to multiple hydrogel compositions and scalable to the number of zones patterned. We confirmed that the methodology to create distinct zones resulted in a continuous, annealed hydrogel with regional interfaces that did not hinder the transport of soluble molecules. Further, the interface between hydrogel regions did not disrupt microchannel structure, epithelial lumen formation, or media perfusion through an acellular or cellularized microchannel. Finally, we demonstrated spatially patterned tubulogenic sprouting of a continuous epithelial tube into the surrounding hydrogel confined to local regions with stromal cell populations, illustrating spatial control of cell-cell interactions and signaling gradients. This easy-to-use system has wide utility for modeling three-dimensional epithelial and endothelial tissue interactions with heterogeneous hydrogel compositions and/or stromal cell populations to investigate their mechanistic roles during development, homeostasis, or disease.
{"title":"Zonal Patterning of Extracellular Matrix and Stromal Cell Populations Along a Perfusable Cellular Microchannel","authors":"Brea Chernokal, Bryan J Ferrick, Jason P Gleghorn","doi":"10.1039/d4lc00579a","DOIUrl":"https://doi.org/10.1039/d4lc00579a","url":null,"abstract":"The spatial organization of biophysical and biochemical cues in the extracellular matrix (ECM) in concert with reciprocal cell-cell signaling is vital to tissue patterning during development. However, elucidating the role an individual microenvironmental factor plays using existing textit{in vivo} models is difficult due to their inherent complexity. In this work, we have developed a microphysiological system to spatially pattern the biochemical, biophysical, and stromal cell composition of the ECM along an epithelialized 3D microchannel. This technique is adaptable to multiple hydrogel compositions and scalable to the number of zones patterned. We confirmed that the methodology to create distinct zones resulted in a continuous, annealed hydrogel with regional interfaces that did not hinder the transport of soluble molecules. Further, the interface between hydrogel regions did not disrupt microchannel structure, epithelial lumen formation, or media perfusion through an acellular or cellularized microchannel. Finally, we demonstrated spatially patterned tubulogenic sprouting of a continuous epithelial tube into the surrounding hydrogel confined to local regions with stromal cell populations, illustrating spatial control of cell-cell interactions and signaling gradients. This easy-to-use system has wide utility for modeling three-dimensional epithelial and endothelial tissue interactions with heterogeneous hydrogel compositions and/or stromal cell populations to investigate their mechanistic roles during development, homeostasis, or disease.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"31 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-10-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142452416","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Abigail G. Ayers, Christia M. Victoriano and Samuel K. Sia
Sample preparation presents a major challenge in point-of-care (POC) diagnostic assays, including ones requiring whole blood as the starting specimen. This study presents an integrated sample preparation device – which we call PRECISE – that performs both plasma separation and nucleic acid extraction, enabling streamlined sample preparation from whole blood requiring only a commercially available blood collection tool and a syringe, and no other external equipment or electricity. Plasma separation is performed using a dual-membrane filter (which filters out blood components while limiting membrane clogging) integrated into the cartridge, and nucleic acid extraction is performed by users moving magnets (to mix the samples, and along a guided track). The plasma filtration demonstrated recovery on par with lab-based centrifugation, and the extraction module showed performance similar to benchtop-based magnetic bead extraction. A sample-to-result demonstration on 50 μL of whole blood spiked with virions of hepatitis C virus (HCV), operating the PRECISE cartridge in 16 minutes followed by benchtop PCR, showed a limit of detection (∼6770 IU mL−1) on the order of the minimal requirements of target product profile for POC HCV detection. Future work on the PRECISE cartridge, building on POC accessibility and fast sample preparation demonstrated in this work, may enable detection of bloodborne pathogens from whole-blood specimens collected at the POC.
{"title":"Integrated device for plasma separation and nucleic acid extraction from whole blood toward point-of-care detection of bloodborne pathogens†","authors":"Abigail G. Ayers, Christia M. Victoriano and Samuel K. Sia","doi":"10.1039/D4LC00571F","DOIUrl":"10.1039/D4LC00571F","url":null,"abstract":"<p >Sample preparation presents a major challenge in point-of-care (POC) diagnostic assays, including ones requiring whole blood as the starting specimen. This study presents an integrated sample preparation device – which we call PRECISE – that performs both plasma separation and nucleic acid extraction, enabling streamlined sample preparation from whole blood requiring only a commercially available blood collection tool and a syringe, and no other external equipment or electricity. Plasma separation is performed using a dual-membrane filter (which filters out blood components while limiting membrane clogging) integrated into the cartridge, and nucleic acid extraction is performed by users moving magnets (to mix the samples, and along a guided track). The plasma filtration demonstrated recovery on par with lab-based centrifugation, and the extraction module showed performance similar to benchtop-based magnetic bead extraction. A sample-to-result demonstration on 50 μL of whole blood spiked with virions of hepatitis C virus (HCV), operating the PRECISE cartridge in 16 minutes followed by benchtop PCR, showed a limit of detection (∼6770 IU mL<small><sup>−1</sup></small>) on the order of the minimal requirements of target product profile for POC HCV detection. Future work on the PRECISE cartridge, building on POC accessibility and fast sample preparation demonstrated in this work, may enable detection of bloodborne pathogens from whole-blood specimens collected at the POC.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 22","pages":" 5124-5136"},"PeriodicalIF":6.1,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d4lc00571f?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142448115","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Inês M. Gonçalves, Muhammad Afzal, Nithil Kennedy, Ana Moita, Rui Lima, Serge Ostrovidov, Takeshi Hori, Yuji Nashimoto, Hirokazu Kaji
One of the most complex human physiological processes to study is pregnancy. Standard animal models, as well as two-dimensional models, lack the complexity and biological relevance required to accurately study such a physiological process. Recent studies have focused on the development of three-dimensional models based on microfluidic systems, designated as placental microphysiological systems (PMPSs). PMPS devices provide a model of the placental barrier through culturing relevant cell types in specific arrangements and media to mimic the in vivo environment of the maternal–fetal circulation. Here, recent developments of PMPS models for embryo uterine implantation, preeclampsia evaluation, and toxicological screening are presented. Studies that use bioprinting techniques are also discussed. Lastly, recent developments in endometrium microphysiological systems are reviewed. All these presented models showed their superiority compared to standard models in recapitulating the biological environment seen in vivo. However, several limitations regarding the types of cells and materials used for these systems were also widely reported. Despite the need for further improvements, PMPS models contribute to a better understanding of the biological mechanisms surrounding pregnancy and the respective pathologies.
{"title":"Placental microphysiological systems: new advances on promising platforms that mimic the microenvironment of the human placenta","authors":"Inês M. Gonçalves, Muhammad Afzal, Nithil Kennedy, Ana Moita, Rui Lima, Serge Ostrovidov, Takeshi Hori, Yuji Nashimoto, Hirokazu Kaji","doi":"10.1039/d4lc00500g","DOIUrl":"https://doi.org/10.1039/d4lc00500g","url":null,"abstract":"One of the most complex human physiological processes to study is pregnancy. Standard animal models, as well as two-dimensional models, lack the complexity and biological relevance required to accurately study such a physiological process. Recent studies have focused on the development of three-dimensional models based on microfluidic systems, designated as placental microphysiological systems (PMPSs). PMPS devices provide a model of the placental barrier through culturing relevant cell types in specific arrangements and media to mimic the <em>in vivo</em> environment of the maternal–fetal circulation. Here, recent developments of PMPS models for embryo uterine implantation, preeclampsia evaluation, and toxicological screening are presented. Studies that use bioprinting techniques are also discussed. Lastly, recent developments in endometrium microphysiological systems are reviewed. All these presented models showed their superiority compared to standard models in recapitulating the biological environment seen <em>in vivo</em>. However, several limitations regarding the types of cells and materials used for these systems were also widely reported. Despite the need for further improvements, PMPS models contribute to a better understanding of the biological mechanisms surrounding pregnancy and the respective pathologies.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"18 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142443900","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Saranath Sripriya, Cyril Picard, Vincent Larrey, Frank Fournel, Elisabeth Charlaix
The energy of saline gradients is a very promising source of non-intermittent renewable energy, the exploitation of which is hampered by the lack of viable technology. The most investigated harvesting methods rely on selective transport of ions or water molecules through semi-permeable or ion-selective membranes, which demonstrate limited power densities of the order of a few W/m2. While in the last decade single nanofluidic objects such as nanopores of nanotubes have opened up very promising prospects with power density capabilities in the kW or even MW/m2, scale-up efforts face serious issues, as concentration polarization phenomena result in a massive loss of performance. We propose here a concept of nanofluidic exchanger for power generation from saline gradients, focused on designing a nanoscale flow able to harvest the power at the output of the nanopores. We study analytically and numerically a simple exchanger made of a selective nanoslit fed by a nanofluidic assembly. One specific feature of such an exchanger relies on the non-linear ion fluxes through the nanoslit analytically expressed from the integration of the Poison-Nernt Planck equation. Such an elemental brick could be massively parallelized in stackable electricity-generating layers using standard technologies of the semi-conductors industry. We demonstrate here a scheme for rationalizing the choice of the exchanger parameters, taking into account the transport properties at all scales. The full numerical resolution of three-dimensional device shows that net power densities of 300 W/m2 and more can be achieved.
{"title":"A nanofluidic exchanger for harvesting saline gradients energy","authors":"Saranath Sripriya, Cyril Picard, Vincent Larrey, Frank Fournel, Elisabeth Charlaix","doi":"10.1039/d4lc00544a","DOIUrl":"https://doi.org/10.1039/d4lc00544a","url":null,"abstract":"The energy of saline gradients is a very promising source of non-intermittent renewable energy, the exploitation of which is hampered by the lack of viable technology. The most investigated harvesting methods rely on selective transport of ions or water molecules through semi-permeable or ion-selective membranes, which demonstrate limited power densities of the order of a few W/m2. While in the last decade single nanofluidic objects such as nanopores of nanotubes have opened up very promising prospects with power density capabilities in the kW or even MW/m2, scale-up efforts face serious issues, as concentration polarization phenomena result in a massive loss of performance. We propose here a concept of nanofluidic exchanger for power generation from saline gradients, focused on designing a nanoscale flow able to harvest the power at the output of the nanopores. We study analytically and numerically a simple exchanger made of a selective nanoslit fed by a nanofluidic assembly. One specific feature of such an exchanger relies on the non-linear ion fluxes through the nanoslit analytically expressed from the integration of the Poison-Nernt Planck equation. Such an elemental brick could be massively parallelized in stackable electricity-generating layers using standard technologies of the semi-conductors industry. We demonstrate here a scheme for rationalizing the choice of the exchanger parameters, taking into account the transport properties at all scales. The full numerical resolution of three-dimensional device shows that net power densities of 300 W/m2 and more can be achieved.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"18 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142440003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alexis Lefevre, Cristian Brandi, Adele De Ninno, Filippo Ruggiero, Enrico Verona, Michaël Gauthier, Paolo Bisegna, Aude Bolopion and Federica Caselli
This work presents an innovative all-electrical platform for selective single-particle manipulation. The platform combines microfluidic impedance cytometry for label-free particle characterization and dielectrophoresis for contactless multi-way particle separation. The microfluidic chip has a straightforward coplanar electrode layout and no particle pre-focusing mechanism is required. An original online algorithm analyzes the impedance signals of each incoming particle and regulates in real time the dielectrophoretic voltages according to a desired control logic. As a proof-of-concept, three operation modes are demonstrated on a mixture of 8, 10, and 12 μm diameter beads: (i) particle position swapping across the channel axis, irrespective of particle size, (ii) size-based particle separation, irrespective of particle position, and (iii) sorting of a selected sequence of particles. As a perspective, the versatility of impedance cytometry and dielectrophoresis, and the possibility of configuring alternative control logics, hold promise for advanced particle and cell manipulation.
{"title":"Real-time impedance-activated dielectrophoretic actuation for reconfigurable manipulation of single flowing particles†","authors":"Alexis Lefevre, Cristian Brandi, Adele De Ninno, Filippo Ruggiero, Enrico Verona, Michaël Gauthier, Paolo Bisegna, Aude Bolopion and Federica Caselli","doi":"10.1039/D4LC00622D","DOIUrl":"10.1039/D4LC00622D","url":null,"abstract":"<p >This work presents an innovative all-electrical platform for selective single-particle manipulation. The platform combines microfluidic impedance cytometry for label-free particle characterization and dielectrophoresis for contactless multi-way particle separation. The microfluidic chip has a straightforward coplanar electrode layout and no particle pre-focusing mechanism is required. An original online algorithm analyzes the impedance signals of each incoming particle and regulates in real time the dielectrophoretic voltages according to a desired control logic. As a proof-of-concept, three operation modes are demonstrated on a mixture of 8, 10, and 12 μm diameter beads: (i) particle position swapping across the channel axis, irrespective of particle size, (ii) size-based particle separation, irrespective of particle position, and (iii) sorting of a selected sequence of particles. As a perspective, the versatility of impedance cytometry and dielectrophoresis, and the possibility of configuring alternative control logics, hold promise for advanced particle and cell manipulation.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 22","pages":" 5145-5154"},"PeriodicalIF":6.1,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142431705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
With advancements in human induced pluripotent stem cell (hiPSC) technology, there is an increasing demand for quality control techniques to manage the long-term process of target cell production effectively. While monitoring systems designed for use within incubators are promising for assessing culture quality, existing systems still face challenges in terms of compactness, throughput, and available metrics. To address these limitations, we have developed a compact and high-throughput lens-free imaging device named INSPCTOR. The device is as small as a standard culture plate, which allows for the installation of multiple units within an incubator. INSPCTOR utilises a large thin-film transistor image sensor, enabling simultaneous observation of six independent culture environments, each approximately 1 cm2. With this device, we successfully monitored the confluency of hiPSC cultures and identified the onset timing of epithelial-to-mesenchymal transition during mesodermal induction. Additionally, we quantified the beating frequency and conduction of hiPSC-derived cardiomyocytes by using high-speed imaging modes. This enabled us to identify the onset of spontaneous beating during differentiation and assess chronotropic responses in drug evaluations. Moreover, by tracking beating frequency over 10 days of cardiomyocyte maturation, we identified week-scale and daily-scale fluctuations, the latter of which correlated with cellular metabolic activity. The metrics derived from this device would enhance the reproducibility and quality of target cell production.
{"title":"Compact lens-free imager using thin-film transistor for long-term quantitative monitoring of stem cell culture and cardiomyocyte production","authors":"Taishi Kakizuka, Tohru Natsume, Takeharu Nagai","doi":"10.1039/d4lc00528g","DOIUrl":"https://doi.org/10.1039/d4lc00528g","url":null,"abstract":"With advancements in human induced pluripotent stem cell (hiPSC) technology, there is an increasing demand for quality control techniques to manage the long-term process of target cell production effectively. While monitoring systems designed for use within incubators are promising for assessing culture quality, existing systems still face challenges in terms of compactness, throughput, and available metrics. To address these limitations, we have developed a compact and high-throughput lens-free imaging device named INSPCTOR. The device is as small as a standard culture plate, which allows for the installation of multiple units within an incubator. INSPCTOR utilises a large thin-film transistor image sensor, enabling simultaneous observation of six independent culture environments, each approximately 1 cm<small><sup>2</sup></small>. With this device, we successfully monitored the confluency of hiPSC cultures and identified the onset timing of epithelial-to-mesenchymal transition during mesodermal induction. Additionally, we quantified the beating frequency and conduction of hiPSC-derived cardiomyocytes by using high-speed imaging modes. This enabled us to identify the onset of spontaneous beating during differentiation and assess chronotropic responses in drug evaluations. Moreover, by tracking beating frequency over 10 days of cardiomyocyte maturation, we identified week-scale and daily-scale fluctuations, the latter of which correlated with cellular metabolic activity. The metrics derived from this device would enhance the reproducibility and quality of target cell production.","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":"85 1","pages":""},"PeriodicalIF":6.1,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142431706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chunqiu Zhang, Ning Rong, Ziyi Lin, Peng-Qi Li, Jingyao Shi, Wei Zhou, Lili Niu, Fei Li, Rongxin Tang, Lei Li and Long Meng
Assisted reproductive technology (ART) has emerged as a crucial method in modern medicine for tackling infertility. However, the success of fertilization depends on the quality and quantity of sperm, often necessitating invasive surgical intervention, which presents challenges for non-invasive in vitro fertilization. Acoustic microfluidics technology has found widespread application across various biological contexts. In this paper, we propose to introduce a novel approach using asymmetric acoustic streaming generated by a single interdigital transducer (IDT) to enhance sperm concentration and improve fertilization in vitro, particularly in cases of moderate oligozoospermia. The concentration of particles increased approximately 6-fold in the central region after acoustic enrichment. Moreover, sperm motility was significantly improved without additional DNA fragmentation, and all the oocytes remained viable after 5 min of acoustic enrichment. Notably, acoustic enrichment accelerated fertilization and embryo development, leading to a higher fertilization rate and faster cleavage speed. Specifically, within 36 hours, the multiple-cell embryo ratio was significantly increased compared to the control group. This finding further validates the feasibility and non-invasiveness of acoustic enrichment for sperm fertilization in vitro. This work provides a promising tool for in vitro fertilization, holding significant implications for assisted reproduction.
辅助生殖技术(ART)已成为现代医学解决不孕不育问题的重要方法。然而,受精成功与否取决于精子的质量和数量,通常需要进行侵入性手术干预,这给无创体外受精带来了挑战。声学微流控技术已在各种生物领域得到广泛应用。在本文中,我们提出了一种新方法,利用单个趾间换能器(IDT)产生的不对称声流来提高精子浓度,改善体外受精,尤其是在中度少精症的情况下。声波富集后,中央区域的颗粒浓度增加了约 6 倍。此外,精子的运动能力也得到了明显改善,而且不会造成额外的 DNA 断裂,所有卵母细胞在声学富集 5 分钟后仍能存活。值得注意的是,声学富集加速了受精和胚胎发育,使受精率更高,裂殖速度更快。具体来说,与对照组相比,36 小时内多细胞胚胎比率明显增加。这一发现进一步验证了声学富集技术在体外精子受精方面的可行性和非侵入性。这项工作为体外受精提供了一种前景广阔的工具,对辅助生殖具有重要意义。
{"title":"Acoustic enrichment of sperm for in vitro fertilization†","authors":"Chunqiu Zhang, Ning Rong, Ziyi Lin, Peng-Qi Li, Jingyao Shi, Wei Zhou, Lili Niu, Fei Li, Rongxin Tang, Lei Li and Long Meng","doi":"10.1039/D4LC00604F","DOIUrl":"10.1039/D4LC00604F","url":null,"abstract":"<p >Assisted reproductive technology (ART) has emerged as a crucial method in modern medicine for tackling infertility. However, the success of fertilization depends on the quality and quantity of sperm, often necessitating invasive surgical intervention, which presents challenges for non-invasive <em>in vitro</em> fertilization. Acoustic microfluidics technology has found widespread application across various biological contexts. In this paper, we propose to introduce a novel approach using asymmetric acoustic streaming generated by a single interdigital transducer (IDT) to enhance sperm concentration and improve fertilization <em>in vitro</em>, particularly in cases of moderate oligozoospermia. The concentration of particles increased approximately 6-fold in the central region after acoustic enrichment. Moreover, sperm motility was significantly improved without additional DNA fragmentation, and all the oocytes remained viable after 5 min of acoustic enrichment. Notably, acoustic enrichment accelerated fertilization and embryo development, leading to a higher fertilization rate and faster cleavage speed. Specifically, within 36 hours, the multiple-cell embryo ratio was significantly increased compared to the control group. This finding further validates the feasibility and non-invasiveness of acoustic enrichment for sperm fertilization <em>in vitro</em>. This work provides a promising tool for <em>in vitro</em> fertilization, holding significant implications for assisted reproduction.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 22","pages":" 5113-5123"},"PeriodicalIF":6.1,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d4lc00604f?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142405505","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Shashwat S. Agarwal, Marcos Cortes-Medina, Jacob C. Holter, Alex Avendano, Joseph W. Tinapple, Joseph M. Barlage, Miles M. Menyhert, Lotanna M. Onua and Jonathan W. Song
Blood and lymphatic vessels in the body are central to molecular and cellular transport, tissue repair, and pathophysiology. Several approaches have been employed for engineering microfabricated blood and lymphatic vessels in vitro, yet traditionally these approaches require specialized equipment, facilities, and research training beyond the capabilities of many biomedical laboratories. Here we present xurography as an inexpensive, accessible, and versatile rapid prototyping technique for engineering cylindrical and lumenized microvessels. Using a benchtop xurographer, or a cutting plotter, we fabricated modular multi-layer poly(dimethylsiloxane) (PDMS)-based microphysiological systems (MPS) that house endothelial-lined microvessels approximately 260 μm in diameter embedded within a user-defined 3-D extracellular matrix (ECM). We validated the vascularized MPS (or vessel-on-a-chip) by quantifying changes in blood vessel permeability due to the pro-angiogenic chemokine CXCL12. Moreover, we demonstrated the reconfigurable versatility of this approach by engineering a total of four distinct vessel-ECM arrangements, which were obtained by only minor adjustments to a few steps of the fabrication process. Several of these arrangements, such as ones that incorporate close-ended vessel structures and spatially distinct ECM compartments along the same microvessel, have not been widely achieved with other microfabrication strategies. Therefore, we anticipate that our low-cost and easy-to-implement fabrication approach will facilitate broader adoption of MPS with customizable vascular architectures and ECM components while reducing the turnaround time required for iterative designs.
{"title":"Rapid low-cost assembly of modular microvessel-on-a-chip with benchtop xurography†","authors":"Shashwat S. Agarwal, Marcos Cortes-Medina, Jacob C. Holter, Alex Avendano, Joseph W. Tinapple, Joseph M. Barlage, Miles M. Menyhert, Lotanna M. Onua and Jonathan W. Song","doi":"10.1039/D4LC00565A","DOIUrl":"10.1039/D4LC00565A","url":null,"abstract":"<p >Blood and lymphatic vessels in the body are central to molecular and cellular transport, tissue repair, and pathophysiology. Several approaches have been employed for engineering microfabricated blood and lymphatic vessels <em>in vitro</em>, yet traditionally these approaches require specialized equipment, facilities, and research training beyond the capabilities of many biomedical laboratories. Here we present xurography as an inexpensive, accessible, and versatile rapid prototyping technique for engineering cylindrical and lumenized microvessels. Using a benchtop xurographer, or a cutting plotter, we fabricated modular multi-layer poly(dimethylsiloxane) (PDMS)-based microphysiological systems (MPS) that house endothelial-lined microvessels approximately 260 μm in diameter embedded within a user-defined 3-D extracellular matrix (ECM). We validated the vascularized MPS (or vessel-on-a-chip) by quantifying changes in blood vessel permeability due to the pro-angiogenic chemokine CXCL12. Moreover, we demonstrated the reconfigurable versatility of this approach by engineering a total of four distinct vessel-ECM arrangements, which were obtained by only minor adjustments to a few steps of the fabrication process. Several of these arrangements, such as ones that incorporate close-ended vessel structures and spatially distinct ECM compartments along the same microvessel, have not been widely achieved with other microfabrication strategies. Therefore, we anticipate that our low-cost and easy-to-implement fabrication approach will facilitate broader adoption of MPS with customizable vascular architectures and ECM components while reducing the turnaround time required for iterative designs.</p>","PeriodicalId":85,"journal":{"name":"Lab on a Chip","volume":" 21","pages":" 5065-5076"},"PeriodicalIF":6.1,"publicationDate":"2024-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2024/lc/d4lc00565a?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142383747","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}