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We demonstrate a microscopy technique for precision sensing across a wide field of view with high spatial precision through pulsed electron spin resonance of nitrogen vacancy centers in diamond.
Localization of a single fluorescent particle with sub-diffraction limit accuracy is a key merit in fluorescence microscopy. Implementation of nonlinear filtering algorithms prior the localization process can improve the localization accuracy of standard existing methods and also enable the localization of overlapping particles, allowing the use of increased fluorophore activation density, and thereby...
Fluorescence apertureless near-field optical microscopy (FASNOM) gained interest from researchers in the last years because of its promising capabilities: sub-wavelength resolution and ability to exploit the chemical sensitivity of fluorescent tags. FASNOM exploits the fact that metallic structures in close proximity of a fluorophore or a photoluminescent sample have complex effects on fluorescence,...
Scanless two-photon microscopy uses a spatial light modulator (SLM) to shape the incoming laser beam into any user-defined light pattern. These microscopes, that do not contain mechanically moving parts, improve the temporal resolution of conventional scanning multiphoton microscopy since they highly mitigate the severe temporal limitations inherent to sequential scanning of the sample [1]. However,...
The human immunodeficiency virus (HIV) is the agent of the global epidemic of the immune disease AIDS (acquired immunodeficiency syndrome). Therefore further investigations on the viral structure and molecular mechanisms are of particular importance in the search for medical therapies.
Single- and few-layer graphene was studied with simultaneous third-harmonic and multiphoton-induced fluorescence microscopy. Both CVD grown and exfoliated graphene were studied [1,2]. The photo-thermal CVD growth was carried out on copper foils in a methane/hydrogen atmosphere, followed by a transfer process. Oxidized silicon substrates were used as the final substrate in both cases. The oxide thickness...
Overcoming the spatial resolution limitations in optical microscopy due to the diffraction of light has become a major challenge in modern microscopy. To overcome this limit, we demonstrate here the upconversion process in rare-earth-doped fluoride nanoparticles (RE:NPs) that may serve as novel sources for fluorescent microscopy. RE:NPs have been reported to serve as efficient biolabels for cellular...
We build a portable, low-cost microscope capable of bright-field and fluorescence imaging with micron-scale resolution in reflection or transmission mode. Software control of image capture and actuation enables automated focusing and large-area sample visualization.
We demonstrate the first proof of concept of a fiber-based STED nanoscopy illumination system. The fiber yields naturally co-aligned vortex (dark-spot size∼198nm) and Gaussian beams (size∼340nm), potentially enabling sub-30nm resolution imaging.
Super-resolution microscopy with stochastically blinking fluorophores (FPALM/PALM/STORM, etc.) achieves ∼25 nm resolution by localizing molecules in thousands of camera frames. Here I present a solution to speed up recording dramatically, enabling nanoscopy at video rate.
Experiments were conducted to develop a microscale spatial measurement technique applicable to transparent fluid flows, with the target application being the determination of liquid geometries in multiphase flows. Air/isopropanol interfaces were used to simulate multiphase boundaries and were studied by examining menisci in capillary tubes. Fluorescent seeding particles of 0.5 µm diameter were added...
The presentation gives an introduction and overview of our recently developed Stochastic Optical Fluctuation Imaging microscopy technique. The theoretical foundations are given and the method is exemplified on several model systems. Its advantages and limitations are discussed, and many applications of SOFI in cell microscopy are presented.
Three-dimensional single-molecule localization is a fundamental problem in biological and biophysical experiments such as super-resolution microscopy. Point spread function engineering accompanied by corresponding reconstruction procedures provides a unique approach to increase localization precision. Joint optical/digital design and image reconstruction are investigated.
We demonstrate large field-of-view (FOV) (30 mm2) fluorescence microscopy with submicron resolution using a refractive microlens array. The large area of the array decreases image acquisition time over FOVs by avoiding long-range mechanical scanning.
We present a new approach for optimizing a 3D non-paraxial volume for multiphoton florescence microscopy. Our optimized solutions demonstrate volume reduction of up to 6.5, compared to the best current design for three-photon microscopy.
We demonstrate, for the first time, a multimodal multiphoton microscope using chirped intensity modulation to construct line images from a single element detector.
A recognized advantage of optical microscopy lies in the fact that allows non-invasive three-dimensional (3D) imaging of live cells at the submicron scale with high specificity [1]. The advent of the visible fluorescent proteins [2] and of a myriad of fluorescent tags pushed fluorescence microscopy to become the most popular imaging tool in cell biology. The confocal and multiphoton versions of fluorescence...
Structured illumination microscopy (SIM) using grating excitation can be used to extend the bandwidth of fluorescent microscopy by approximately a factor of 2 in the linear regime. If some of the fluorescent molecules are saturated even greater improvements in resolution are possible; this may, however, lead to high levels of photobleaching and phototoxicity. In this paper we present preliminary results...
We report on recent development of surface plasmon polaritons (SPP) excited by a highly focused radially polarized optical vortex (RPOV) beam on a metal surface and its new applications in surface plasmon resonance (SPR) sensing and imaging. The proposed method reveals a direct phase manipulation through an incident structured beam to its counterpart in SPP with dynamic, reconfigurable and high-efficiency...
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