DISPLACEMENT CURRENT IN BIOLOGICAL MEMBRANES

The functioning of some important membrane bound enzymes of the living systems is connected with charge translocation. A method has been elaborated to measure the displacement current due to the motion of protons on light excitation in the bacteriorhodopsin molecules. From the time distribution and the amplitudes of the current it has been established that the translocation of protons takes five different steps all with different distances inside the proteins

Light Sailboats: Laser driven autonomous microrobots

We introduce a system of light driven microscopic autonomous moving particles that move on a flat surface. The design is simple, yet effective: Micrometer sized objects with wedge shape are produced by photopolymerization, they are covered with a reflective surface. When the area of motion is illuminated perpendicularly from above, the light is deflected to the side by the wedge shaped objects, in the direction determined by the position and orientation of the particles. The momentum change during reflection provides the driving force for an effectively autonomous motion. The system is an efficient tool to study self propelled microscopic robots

Holographic multi-focus 3D two-photon polymerization with real-time calculated holograms.

Two-photon polymerization enables the fabrication of micron sized structures with submicron resolution. Spatial light modulators (SLM) have already been used to create multiple polymerizing foci in the photoresist by holographic beam shaping, thus enabling the parallel fabrication of multiple microstructures. Here we demonstrate the parallel two-photon polymerization of single 3D microstructures by multiple holographically translated foci. Multiple foci were created by phase holograms, which were calculated real-time on an NVIDIA CUDA GPU, and displayed on an electronically addressed SLM. A 3D demonstrational structure was designed that is built up from a nested set of dodecahedron frames of decreasing size. Each individual microstructure was fabricated with the parallel and coordinated motion of 5 holographic foci. The reproducibility and the high uniformity of features of the microstructures were verified by scanning electron microscopy

Biológiai membránok elektromos térszerkezetének vizsgálata = Investigation of the charge structure of biological membranes

A pályázat célja a biológiai membránok fizikai tulajdonságainak vizsgálata volt, különös tekintettel a töltéseloszlás és - ezzel összefüggésben - az elektromos térszerkezet funkcionális szerepére. Modellobjektumunk a protonpumpáló bakteriorodopszin (bR) fehérjét kristályos rendben tartalmazó bíbormembrán. Kutatásainkat és a velük kapcsolatos módszertani fejlesztőmunkát három téma köré csoportosítottuk: 1. A molekulán belüli elektromos töltéseloszlás funkcionális leírása 2. A határfelületi vízréteg szerepe a fehérjék szerkezetében és működésében 3. A bR nemlineáris optikai tulajdonságainak jellemzése Eredmények: 1.Megmutattuk, hogy az általunk kifejlesztett módszer a molekula működését kísérő szerkezetváltozásokat érzékenyen jellemzi, és lehetőséget teremt az elektromos töltésátrendeződések funkcionális értelmezésére. 2.Rámutattunk, hogy a semleges sók anionjainak tulajdonított Hofmeister-effektus a fehérje-víz határfelületi réteg szerkezetváltozásaival magyarázható. Elméletünk elsőként ad teljeskörű kvalitatív leírást a 120 éves problémakörre. 3.Integrált optikai módszerekkel az eddigieknél érzékenyebben detektáltuk a bR molekulában - a lokális elektromos tér változásai következtében ? fellépő abszorpció- és törésmutató-változásokat. Megmutattuk, hogy a molekula gerjesztését követő spektrális változások alkalmasak fénykapcsolásra és -modulálásra. Az eredmények gyakorlati alkalmazása az optoelektronikában és a bioszenzorikában várható. | In the framework of the project, we investigated some physical properties of biological membranes, with special respect to the physiological role of the electric charge distribution (hence the electric field structure). Our model object was purple membrane, containing the paradigmatic proton pumping molecule bacteriorhodopsin (bR) in a quasi crystalline structure. Our research work and the related methodological developments were centered round three topics: 1.Functional description of the intra-molecular charge distribution 2.The role of interfacial water in the structure and function of proteins 3.Characterization of the nonlinear optical properties of bR Results: 1.We showed that our method represents a sensitive test for the characterization of structural changes associated to molecular function, and allows interpretation of charge rearrangements. 2.We pointed out that the Hofmeister effect, attributed to the anions of neutral salts, can be interpreted according to structural changes in the protein-water interface. Our theory gives a full-spectrum qualitative description, solving a 120-years old puzzle. 3.Using integrated optical (IO) techniques, we could sensitively characterize the nonlinear optical properties of bR films, and showed that IO light switching and modulation can be performed based on the light-induced spectral changes of bR. Practical utilization of the results is expected in optoelectronics and sensorics

Mikromanipulációs kísérletek lézercsipesszel = Micromanipulation experiments with optical tweezers

A kutatás az optikai mikromanipuláció területén, különböző irányokban végzett fejlesztéseket és kutatási alkalmazásokat képviselt. A mikromanipuláció területén azt vizsgáltuk, milyen új manipulációs lehetőségeket nyújtanak speciális alakú próbatestek (ellentétben az általáéban használt gömb alakkal). Ennek során kidolgoztuk a lézeres fotopolimerizációs struktúra építés technikáját. Ezzel egyrészt a mikromanipuláció új lehetőségeit vizsgáltuk. Például, lehetőség nyílik torziós manipulálásra, csavarásra, meghatároztuk a DNS molekula torziós rugalmasságát. Ezen kívül, bonyolult struktúrákat építettünk, fénnyel hajtott mikrogépeket, mikrofluidikai csatornákat, integrált optikai szenzorokat. Ezekből bonyolultab összetett rendszereket raktunk össze: fénnyel vezérelt fluorescencia aktivált optikai sejtszeparátort. Kidolgoztuk a fénnyel vezérelt elektroozmózis eljárását. Itt elektromos térrel mozgatott folyadék áramlását vezéreljük fénnyel, ez érdekes jelenség, és új lehetőségeket nyújt mikrofluidikai rendszerek vezérlésében. | The research represented studies in the area of optical micromanipulation, both developing new procedures and basic applications. In the area of micromanipulation we investigated, what new possibilities are offered by test objects of special shapes (as opposed to the generally used spheres). In this process we developed the structure building by laser induced photopolymerization. With this we studied new possibilities of optical manipulation. For example, a poissibility emerges for torsional manipulation, twisting, we determined the torsional elasticity of DNA molecules. In addition, we built complex structures, light driven micromachines, microfluidics channels, integrated optical sensors. From this we constructed complex systems, e.g. a fluorescence activated cell separator. We developed the method of optically controlled electroosmosis. Here the fluid is driven by electric field and this is controlled by light. This is an interesting phenomenon, and is opens new possibilities in the control of microfluidics systems

Direct writing of optical microresonators in a lab-on-a-chip for label-free biosensing

Whispering gallery mode (WGM) resonators are promising optical structures for microfluidic label-free bio-sensors mainly due to their high sensitivity, but from a practical point of view they present numerous constraints that make their use in real laboratory diagnosis application difficult. Herein we report on a monolithic lab on a chip fabricated by a hybrid femtosecond laser micromachining approach, for label-free biosensing. It consists of a polymer WGM microresonator sensor integrated inside a glass microfluidic chip, presenting a refractive index change sensitivity of 61 nm per RIU. The biosensing capabilities of the device have been demonstrated by exploiting the biotin-streptavidin binding affinity, obtaining a measurable minimum surface density increase of 67 x 10(3) molecules per mu m(2)

Multiview microscopy of single cells through microstructure-based indirect optical manipulation

Fluorescent observation of cells generally suffers from the limited axial resolution due to the elongated point spread function of the microscope optics. Consequently, three-dimensional imaging results in axial resolution that is several times worse than the transversal. The optical solutions to this problem usually require complicated optics and extreme spatial stability. A straightforward way to eliminate anisotropic resolution is to fuse images recorded from multiple viewing directions achieved mostly by the mechanical rotation of the entire sample. In the presented approach, multiview imaging of single cells is implemented by rotating them around an axis perpendicular to the optical axis by means of holographic optical tweezers. For this, the cells are indirectly trapped and manipulated with special microtools made with two-photon polymerization. The cell is firmly attached to the microtool and is precisely manipulated with 6 degrees of freedom. The total control over the cells' position allows for its multiview fluorescence imaging from arbitrarily selected directions. The image stacks obtained this way are combined into one 3D image array with a multiview image processing pipeline resulting in isotropic optical resolution that approaches the lateral diffraction limit. The presented tool and manipulation scheme can be readily applied in various microscope platforms

Microfluidic study of the chemotactic response of Escherichia coli to amino acids, signaling molecules and secondary metabolites

Quorum sensing and chemotaxis both affect bacterial behavior on the population level. Chemotaxis shapes the spatial distribution of cells, while quorum sensing realizes a cell-density dependent gene regulation. An interesting question is if these mechanisms interact on some level: Does quorum sensing, a density dependent process, affect cell density itself via chemotaxis? Since quorum sensing often spans across species, such a feedback mechanism may also exist between multiple species. We constructed a microfluidic platform to study these questions. A flow-free, stable linear chemical gradient is formed in our device within a few minutes that makes it suitable for sensitive testing of chemoeffectors: we showed that the amino acid lysine is a weak chemoattractant for Escherichia coli, while arginine is neutral. We studied the effect of quorum sensing signal molecules of Pseudomonas aeruginosa on E. coli chemotaxis. Our results show that N-(3-oxododecanoyl)-homoserine lactone (oxo-C12-HSL) and N-(butryl)-homoserine lactone (C4-HSL) are attractants. Furthermore, we tested the chemoeffector potential of pyocyanin and pyoverdine, secondary metabolites under a quorum sensing control. Pyocyanin is proved to be a weak attractant while pyoverdine are repellent. We demonstrated the usability of the device in co-culturing experiments, where we showed that various factors released by P. aeruginosa affect the dynamic spatial rearrangement of a neighboring E. coli population, while surface adhesion of the cells is also modulated. © 2015 AIP Publishing LLC

Biological Microscopy with Undetected Photons

Novel imaging techniques utilizing nondegenerate, correlated photon pairs sparked intense interest during the last couple of years among scientists of the quantum optics community and beyond. It is a key property of such "ghost imaging" or "quantum interference" methods that they use those photons of the correlated pairs for imaging that never interacted with the sample, allowing detection in a spectral range different from that of the illumination of the object. Extensive applications of these techniques in spectroscopy and microscopy are envisioned, however, their limited spatial resolution to date has not yet supported real-life microscopic investigations of tiny biological objects. Here we report a modification of the method based on quantum interference by using a seeding laser and confocal scanning, that allows the improvement of the resolution of imaging with undetected photons by more than an order of magnitude, and we also present examples of application in the microscopy of biological samples.Comment: This article has been accepted for publication in IEEE Access, but has not been fully edite