Optofluidics, Microfluidics and Nanofluidics最新文献

{"title":"Laser switching contrast microscopy to monitor free and restricted diffusion inside the cell nucleus","authors":"F. Schmitt, Cornelia Junghans, M. Sturm, C. Keuer, H. Eichler, T. Friedrich","doi":"10.1515/optof-2016-0001","DOIUrl":"https://doi.org/10.1515/optof-2016-0001","url":null,"abstract":"Abstract A novel microscopic technique termed laser switching contrast microscopy (LSCM) allows for the imaging of the dynamics of optically switchable proteins in single cell compartments. We present an application for the monitoring of diffusive properties of single molecules of the photo-switchable fluorescent protein Dreiklang (DRK). LSCM in the cell nucleus of Chinese hamster ovary (CHO) cells cytoplasmically expressing DRK unravels quick diffusive equilibration of the DRK molecules inside the whole cytoplasm and inside the cell nucleus within seconds. The nuclear membrane is also highly permeable for DRK. Inside the nucleus entirely distinct regions are found that only partially enable diffusive protein redistribution with mean square displacement proportional to time while in other regions the mobility of the proteins seems to be restricted. After photo-switching string like patterns of light DRK molecules are observed in the cell nucleus. In addition a fraction of these DRK molecules appears immobile. The findings support recent theories of the cell interior described as a random obstacle model with an additional immobile fraction of DRK. Numerical simulations show that at different illumination intensity and different distance from the laser focus similar patterns for fluorescence recovery might be obtained in spite of strongly varying diffusion constants.","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"96 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122274869","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Diffusion behavior of the fluorescent proteins eGFP and Dreiklang in solvents of different viscosity monitored by fluorescence correlation spectroscopy","authors":"Cornelia Junghans, F. Schmitt, V. Vukojević, T. Friedrich","doi":"10.1515/optof-2016-0004","DOIUrl":"https://doi.org/10.1515/optof-2016-0004","url":null,"abstract":"Abstract Fluorescence correlation spectroscopy relies on temporal autocorrelation analysis of fluorescence intensity fluctuations that spontaneously arise in systems at equilibrium due to molecular motion and changes of state that cause changes in fluorescence, such as triplet state transition, photoisomerization and other photophysical transformations, to determine the rates of these processes. The stability of a fluorescent molecule against dark state conversion is of particular concern for chromophores intended to be used as reference tags for comparing diffusion processes on multiple time scales. In this work, we analyzed properties of two fluorescent proteins, the photoswitchable Dreiklang and its parental eGFP, in solvents of different viscosity to vary the diffusion time through the observation volume element by several orders of magnitude. In contrast to eGFP, Dreiklang undergoes a dark-state conversion on the time scale of tens to hundreds of microseconds under conditions of intense fluorescence excitation, which results in artificially shortened diffusion times if the diffusional motion through the observation volume is sufficiently slowed down. Such photophysical quenching processes have also been observed in FCS studies on other photoswitchable fluorescent proteins including Citrine, from which Dreiklang was derived by genetic engineering. This property readily explains the discrepancies observed previously between the diffusion times of eGFP- and Dreiklang-labeled plasma membrane protein complexes.","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"3 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-01-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130276673","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High-throughput fabrication and calibration of compact high-sensitivity plasmonic lab-on-chip for biosensing","authors":"E. Gazzola, A. Pozzato, G. Ruffato, E. Sovernigo, A. Sonato","doi":"10.1515/optof-2016-0002","DOIUrl":"https://doi.org/10.1515/optof-2016-0002","url":null,"abstract":"Abstract Surface plasmon resonance biosensors have recently known a rapid diffusion in the biological field and a large variety of sensor configurations is currently available. Biological applications are increasingly demanding sensor miniaturization, multiple detection in parallel, temperature-controlled environment and high sensitivity. Indeed, versatile and tunable sensing platforms, together with an accurate biological environment monitoring, could improve the realization of custom biosensing devices applicable to different biological reactions. Here we propose a smart and high throughput fabrication protocol for the realization of a custommicrofluidic plasmonic biochip that could be easily tuned and modified to address different biological applications. The sensor chip here presented shows a high sensing capability, monitored by an accurate signal calibration in the presence of concentration and temperature variation.","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"52 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2016-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127127365","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multimode fibres for micro-endoscopy","authors":"S. Turtaev, I. Leite, T. Čižmár","doi":"10.1515/optof-2015-0004","DOIUrl":"https://doi.org/10.1515/optof-2015-0004","url":null,"abstract":"Abstract There has been a tremendous effort in modern microscopy towards miniaturisation and fibre-based technology, driven by the need to access hostile or difficult environments in situ and in vivo. Most of these rely on reducing the size of endoscopes based on fibre-optic bundles, and systems incorporating microfabricated lenses. Recently, the use of standard multimode optical fibres for lensless microscopy has become possible mainly due to advances in holographic beam shaping. This article reviews the methods and techniques behind this progress paving theway towards minimally invasive in vivo imaging as well as other applications of multimode waveguides including on-chip integration of optical micro-manipulation and numerous other biophotonics techniques.","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"33 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2015-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128919028","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"BANSAI - An optofluidic approach for biomedical analysis","authors":"Markus Knoerzer, C. Prokop, Graciete M. Rodrigues Ribeiro, H. Mayer, Jens Brümmer, A. Mitchell, D. Rabus, C. Karnutsch","doi":"10.1515/optof-2015-0003","DOIUrl":"https://doi.org/10.1515/optof-2015-0003","url":null,"abstract":"Abstract Lab-on-a-chip based portable blood analysis systems would allow point-of-care measurements, e.g. in an ambulance, or in remote areas with no fast access to medical care. Such a systemwould provide much faster information about the health of a patient. Here,we present a system that is based on absorption spectroscopy and uses an organic laser, which is tunable in the visible range. The feasibility of the system is shown with a table-top setup using laboratory equipment. Measurements of human albumin show linear behaviour in a range from 2.5 g/L to 60 g/L. In a consecutive setup the system is implemented on a microfluidic chip and is capable of measuring simultaneously transmitted and side scattered intensities, even with ambient light present. Air-suspended grating couplers on polymers are shown as the first element of a lab-on-a-chip implementation.","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"2 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2015-12-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130001520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Monitoring the diffusion behavior of Na,K-ATPase by fluorescence correlation spectroscopy (FCS) upon fluorescence labelling with eGFP or Dreiklang","authors":"Cornelia Junghans, F. Schmitt, V. Vukojević, T. Friedrich","doi":"10.1515/optof-2015-0001","DOIUrl":"https://doi.org/10.1515/optof-2015-0001","url":null,"abstract":"Abstract Measurement of lateral mobility of membraneembedded proteins in living cells with high spatial and temporal precision is a challenging task of optofluidics. Biological membranes are complex structures, whose physico-chemical properties depend on the local lipid composition, cholesterol content and the presence of integral or peripheral membrane proteins, which may be involved in supramolecular complexes or are linked to cellular matrix proteins or the cytoskeleton. The high proteinto- lipid ratios in biomembranes indicate that membrane proteins are particularly subject to molecular crowding, making it difficult to follow the track of individual molecules carrying a fluorescence label. Novel switchable fluorescence proteins such as Dreiklang [1], are, in principle, promising tools to study the diffusion behavior of individual molecules in situations of molecular crowding due to excellent spectral control of the ON- and OFF-switching process. In this work, we expressed an integral membrane transport protein, the Na,K-ATPase comprising the human α2-subunit carrying an N-terminal eGFP or Dreiklang tag and human β1-subunit, in HEK293T cells and measured autocorrelation curves by fluorescence correlation spectroscopy (FCS). Furthermore,we measured diffusion times and diffusion constants of eGFP and Dreiklang by FCS, first, in aqueous solution after purification of the proteins upon expression in E. coli, and, second, upon expression as soluble proteins in the cytoplasm of HEK293T cells. Our data show that the diffusion behavior of the purified eGFP and Dreiklang in solution as well as the properties of the proteins expressed in the cytoplasm are very similar. However, the autocorrelation curves of eGFP- and Dreiklanglabeled Na,K-ATPase measured in the plasma membrane exhibit marked differences, with the Dreiklang-labeled construct showing shorter diffusion times. This may be related to an additional, as yet unrecognized quenching process that occurs on the same time scale as the diffusion of the labeled complexes through the detection volume (1– 100 ms). Since the origin of this quenching process is currently unclear, care has to be taken when the Dreiklang label is intended to be used in FCS applications.","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"54 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2015-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121054987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lamellar spacing of photosystem II membrane fragments upon dehydration studied by neutron membrane diffraction","authors":"J. Pieper, L. Rusevich, T. Hauß, G. Renger","doi":"10.1515/optof-2015-0005","DOIUrl":"https://doi.org/10.1515/optof-2015-0005","url":null,"abstract":"Abstract The effect of dehydration on the lamellar spacing of photosystem II (PS II) membrane fragments from spinach has been investigated using neutron membrane diffraction at room temperature. The diffraction data reveal a major peak at a scattering vector Q of 0.049 Å−1 at a relative humidity (r.h.) of 90% corresponding to a repeat distance D of about 129 Å. Upon dehydration to 44% r.h., this peak shifts to about 0.060 Å−1 corresponding to a distance of 104.7±2.5 Å. Within experimental error, the latter repeat distance remains almost the same at hydration levels below 44% r.h. indicating that most of the hydration water is removed. This result is consistent with the earlier finding that hydration-induced conformational protein motions in PS II membrane fragments are observed above 44% r.h. and correlated with the onset electron transfer in PS II (Pieper et al. 2008, Eur. Biophys. J. 37: 657–663).","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"6 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2015-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132927673","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrated optical biosensor for rapid detection of bacteria","authors":"A. Mathesz, S. Valkai, Attila Újvárosy, Badri L. Aekbote, O. Sipos, B. Stercz, B. Kocsis, D. Szabo, A. Dér","doi":"10.1515/optof-2015-0002","DOIUrl":"https://doi.org/10.1515/optof-2015-0002","url":null,"abstract":"Abstract In medical diagnostics, rapid detection of pathogenic bacteria from body fluids is one of the basic issues. Most state-of-the-art methods require optical labeling, increasing the complexity, duration and cost of the analysis. Therefore, there is a strong need for developing selective sensory devices based on label-free techniques, in order to increase the speed, and reduce the cost of detection. In a recent paper, we have shown that an integrated optical Mach-Zehnder interferometer, a highly sensitive all-optical device made of a cheap photopolymer, can be used as a powerful lab-on-a-chip tool for specific, labelfree detection of proteins. By proper modifications of this technique, our interferometric biosensor was combined with a microfluidic system allowing the rapid and specific detection of bacteria from solutions, having the surface of the sensor functionalized by bacterium-specific antibodies. The experiments proved that the biosensor was able to detect Escherichia coli bacteria at concentrations of 106 cfu/ml within a few minutes, that makes our device an appropriate tool for fast, label-free detection of bacteria from body fluids such as urine or sputum. On the other hand, possible applications of the device may not be restricted to medical microbiology, since bacterial identification is an important task in microbial forensics, criminal investigations, bio-terrorism threats and in environmental studies, as well.","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"5 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2015-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126774407","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Running Droplet Optical Multiplexer","authors":"L. Brandhoff, M. Akhtar, M. Bülters, R. Bergmann, M. Vellekoop","doi":"10.2478/optof-2014-0007","DOIUrl":"https://doi.org/10.2478/optof-2014-0007","url":null,"abstract":"Abstract We present an optofluidic device for switching light from multiple inputs to one common output. The device uses a microfluidic channel filled with high index of refraction oil as a waveguide, and moves low refractive index interruptions in the form of aqueous droplets through the channel. Whenever a droplet passes one of the optical inputs, this specific input is switched through to the output. This produces a running switching of one output following the other creating a 8x1 multiplexer.","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"3 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2014-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133399454","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chitosan microgels obtained by on-chip crosslinking reaction employing a microfluidic device","authors":"V. Zamora-Mora, D. Velasco, R. Hernández, C. Mijangos","doi":"10.2478/optof-2014-0004","DOIUrl":"https://doi.org/10.2478/optof-2014-0004","url":null,"abstract":"Abstract In the present work, we report on the preparation of microgels of chitosan crosslinked with sodium tripolyphosphate (TPP) employing the microfluidics technique (MF). To achieve this, several flow focusing geometries were designed and tested. As a first step, a two-inlet flow focusing geometry was employed to emulsify chitosan and the crosslinking reaction was carried out offchip. This procedure did not allow separating the resulting chitosan microgels due to an incomplete crosslinking reaction. A crosslinking reaction on-chip was studied as an alternative. A four-inlet flow focusing geometrywas designed in which three dispersed phases, chitosan 0.25% (w/v), TPP 0.05% (w/v) and acetic acid 1% (v/v) and an continuous phase mineral oil + Span 80 (3% w/v) were employed. The flow rates for the continuous phase were varied from 6.7 to 11.7 μL/min and chitosan microgels were successfully obtained with average diameters from 68 to 42 μm. The average size of the microgels outside the MF device decreased up to ~21% with respect to their size inside the MF device due to partial expulsion of water from the microgels when complete gelation occurred.","PeriodicalId":144806,"journal":{"name":"Optofluidics, Microfluidics and Nanofluidics","volume":"474 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2014-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129212234","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}