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| 1 | Converting lateral scanning into axial focusing to speed up three-dimensional microscopy显示文摘In optical microscopy,the slow axial scanning rate of the objective or the sample has traditionally limited the speed of volumetric imaging.Recently,by conjugating either a movable mirror to the image plane in a remote-focusing geometry or an electrically tuneable lens(ETL)to the back focal plane,rapid axial scanning has been achieved.However,mechanical actuation of a mirror limits the axial scanning rate(usually only 10–100 Hz for piezoelectric or voice coil-based actuators),while ETLs introduce spherical and higher-order aberrations that prevent high-resolution imaging.In an effort to overcome these limitations,we introduce a novel optical design that transforms a lateral-scan motion into a spherical aberration-free axial scan that can be used for high-resolution imaging.Using a galvanometric mirror,we scan a laser beam laterally in a remote-focusing arm,which is then back-reflected from different heights of a mirror in the image space.We characterize the optical performance of this remote-focusing technique and use it to accelerate axially swept light-sheet microscopy by an order of magnitude,allowing the quantification of rapid vesicular dynamics in three dimensions.We also demonstrate resonant remote focusing at 12 kHz with a two-photon raster-scanning microscope,which allows rapid imaging of brain tissues and zebrafish cardiac dynamics with diffraction-limited resolution. | Tonmoy Chakraborty Bingying Chen Stephan Daetwyler Bo-Jui Chang Oliver Vanderpoorten Etai Sapoznik Clemens F.Kaminski Tuomas P.J.Knowles Kevin M.Dean Reto Fiolka | 2020 | Light(Science & Applications)2020，9，1： | 4 |
| 2 | Scalable integration of nano-,and microfluidics with hybrid two-photon lithography显示文摘Nanofluidic devices have great potential for applications in areas ranging from renewable energy to human health.A crucial requirement for the successful operation of nanofluidic devices is the ability to interface them in a scalable manner with the outside world.Here,we demonstrate a hybrid two photon nanolithography approach interfaced with conventional mask whole-wafer UV-photolithography to generate master wafers for the fabrication of integrated micro and nanofluidic devices.Using this approach we demonstrate the fabrication of molds from SU-8 photoresist with nanofluidic features down to 230 nm lateral width and channel heights from micron to sub-100 nm.Scanning electron microscopy and atomic force microscopy were used to characterize the printing capabilities of the system and show the integration of nanofluidic channels into an existing microfluidic chip design.The functionality of the devices was demonstrated through super-resolution microscopy,allowing the observation of features below the diffraction limit of light produced using our approach.Single molecule localization of diffusing dye molecules verified the successful imprint of nanochannels and the spatial confinement of molecules to 200 nm across the nanochannel molded from the master wafer.This approach integrates readily with current microfluidic fabrication methods and allows the combination of microfluidic devices with locally two-photon-written nano-sized functionalities,enabling rapid nanofluidic device fabrication and enhancement of existing microfluidic device architectures with nanofluidic features. | Oliver Vanderpoorten Quentin Peter Pavan K.Challa Ulrich F.Keyser Jeremy Baumberg Clemens F.Kaminski Tuomas P.J.Knowles | 2019 | Microsystems & Nanoengineering2019，5，1： | 2 |
| 3 | Sequential storage and release of microdroplets显示文摘Droplet microfluidic methods have opened up the possibility of studying a plethora of phenomena ranging from biological to physical or chemical processes at ultra low volumes and high throughput.A key component of such approaches is the ability to trap droplets for observation,and many device architectures for achieving this objective have been developed.A challenge with such approaches is,however,recovering the droplets following their confinement for applications involving further analysis.Here,we present a device capable of generating,confining and releasing microdroplets in a sequential manner.Through a combination of experimental and computational simulations,we shed light on the key features required for successful droplet storage and retrieval.Moreover,we explore the effect of the flow rate of the continuous phase on droplet release,determining that a critical rate is needed to ensure complete droplet deformation through constrictions holding the droplets in place prior to release.Finally,we find that once released,droplets can be retrieved and collected off chip.The ability to generate,store and sequentially release droplets renders such a device particularly promising for future applications where reactions may not only be monitored on-chip,but droplets can also be retrieved for further analysis,facilitating new exploratory avenues in the fields of analytical chemistry and biology. | Zenon Toprakciogl Tuomas P.J.Knowles | 2021 | Microsystems & Nanoengineering2021，7，5： | 0 |
| 4 | Rapid two-dimensional characterisation of proteins in solution显示文摘Microfluidic platforms provide an excellent basis for working with heterogeneous samples and separating biomolecular components at high throughput,with high recovery rates and by using only very small sample volumes.To date,several micron scale platforms with preparative capabilities have been demonstrated.Here we describe and demonstrate a microfluidic device that brings preparative and analytical operations together onto a single chip and thereby allows the acquisition of multidimensional information.We achieve this objective by using a free-flow electrophoretic separation approach that directs fractions of sample into an on-chip analysis unit,where the fractions are characterised through a microfluidic diffusional sizing process.This combined approach therefore allows simultaneously quantifying the sizes and the charges of components in heterogenous mixtures.We illustrate the power of the platform by describing the size distribution of a mixture comprising components which are close in size and cannot be identified as individual components using state-of-the-art solution sizing techniques on their own.Furthermore,we show that the platform can be used for two-dimensional fingerprinting of heterogeneous protein mixtures within tens of seconds,opening up a possibility to obtain multiparameter data on biomolecular systems on a minute timescale. | Kadi LSaar Quentin Peter Thomas Muller Pavan K.Challa Therese W.Herling Tuomas P.J.Knowles | 2019 | Microsystems & Nanoengineering2019，5，1： | 0 |
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