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We demonstrate sub-wavelength sectioning on biological samples with a conventional confocal microscope. This optical sectioning is achieved by the phenomenon of supercritical angle fuorescence, wherein only a fluorophore next to the…

Recent advances in optical technology have significantly enhanced the resolution of imaging of living cells, achieving nanometer-scale precision. However, the crowded three-dimensional environment within cells presents a challenge for…

Optics · Physics 2024-11-20 Vahid Nasirimarekani , Zuzana Ditte , Eberhard Bodenschatz

Fluorescence detection, either involving propagating or near-field emission, is widely being used in spectroscopy, sensing and microscopy. Total internal reflection fluorescence (TIRF) confines fluorescence excitation by an evanescent…

Quantitative Methods · Quantitative Biology 2020-06-24 Martin Oheim , Adi Salomon , Maia Brunstein

Total internal reflection fluorescence (TIRF) microscopy and its variants are key technologies for visualizing the dynamics of single molecules or organelles in live cells. Yet, truely quantitative TIRF remains problematic. One unknown…

Biological Physics · Physics 2017-06-07 Maia Brunstein , Lopamudra Roy , Martin Oheim

Super-resolution Structured Illumination Microscopy (SR-SIM) enables fluorescence microscopy beyond the diffraction limit at high frame rates. Compared to other super-resolution microscopy techniques, the low photon fluence used in SR-SIM…

We present a new technique for obtaining simultaneous multimodal quantitative phase and fluorescence microscopy of biological cells, providing both quantitative phase imaging and molecular specificity using a single camera. Our system is…

Optics · Physics 2019-04-03 Yoav N. Nygate , Gyanendra Singh , Itay Barnea , Natan T. Shaked

We demonstrate an electro-optic wide-field method to enable fluorescence lifetime microscopy (FLIM) with high throughput and single-molecule sensitivity. Resonantly driven Pockels cells are used to efficiently gate images at 39 MHz,…

Optics · Physics 2021-09-23 Adam J. Bowman , Mark A. Kasevich

We report the development and detailed calibration of a multiphoton fluorescence lifetime imaging system (FLIM) using a streak camera. The present system is versatile with high spatial (0.2 micron) and temporal (50 psec) resolution and…

Quantitative Methods · Quantitative Biology 2009-11-10 R. V. Krishnan , H. Saitoh , H. Terada , V. E. Centonze , B. Herman

Subwavelength imaging by microsphere lenses is a promising label-free super-resolution imaging technique. There is a growing interest to use live cells to replace the widely used non-biological microsphere lenses. In this work, we…

Optics · Physics 2021-07-06 Chunlei Jiang , Hangyu Yue , Bing Yan , Taiji Dong , Xiangyu Cui , Zengbo Wang

Fluorescence microscopy is an important and extensively utilised tool for imaging biological systems. However, the image resolution that can be obtained has a limit as defined through the laws of diffraction. Demand for improved resolution…

Biological Physics · Physics 2007-08-27 James H. Rice

Fluorescence imaging is an essential diagnostic tool in many fields, but diffraction-limited optical imaging at depth is limited by scattering. Here, we present a method based on multiple random illuminations, combined with a computational…

Optics · Physics 2026-03-30 Lei Zhu , Tengfei Wu , Bernhard Rauer , Hilton B. de Aguiar , Sylvain Gigan

Oblique plane microscopy is a method enabling light-sheet fluorescence imaging through a single microscope objective lens by focusing on a tilted plane within the sample. To focus the fluorescence emitted by the oblique plane on a camera,…

This thesis centres on the development of multidimensional fluorescence imaging tools, with a particular emphasis on fluorescence lifetime imaging (FLIM) microscopy for application to biological research. The key aspects of this thesis are…

Instrumentation and Detectors · Physics 2017-08-14 Egidijus Auksorius

Fluorescence fluctuation spectroscopy is applied to study molecules, passing through a small observation volume, usually subjected to diffusive or convective motion in liquid phase. We suggest that such a technique could be used to measure…

Fluorescence microscopy is widely used in biological imaging, however scattering from tissues strongly limits its applicability to a shallow depth. In this work we adapt a methodology inspired from stellar speckle interferometry, and…

Investigation of cell structure is hardly imaginable without bright-field microscopy. Numerous modifications such as depth-wise scanning or videoenhancement make this method being state-of-the-art. This raises a question what maximal…

Image and Video Processing · Electrical Eng. & Systems 2020-10-29 Kirill Lonhus , Renata Rychtarikova , Ganna Platonova , Dalibor Stys

We demonstrate volumetric fluorescence lifetime microscopy (FLIM) using the electro-optic FLIM technique. Images acquired in a selective plane illumination microscope are gated using a Pockels cell driven at 80 MHz, enabling light sheet…

FRET-based approaches are a unique tool for sensing the immediate surroundings and interactions of (bio)molecules. FRET imaging and FLIM (Fluorescence Lifetime Imaging Microscopy) enable the visualization of the spatial distribution of…

Fluorescence imaging is the most widely used method for unveiling the molecular composition of biological specimens. However, the weak optical emission of fluorescent probes and the tradeoff between imaging speed and sensitivity is…

We report the cell biological applications of a recently developed multiphoton fluorescence lifetime imaging microscopy system using a streak camera (StreakFLIM). The system was calibrated with standard fluorophore specimens and was shown…

Cell Behavior · Quantitative Biology 2009-11-10 R. V. Krishnan , Eva Biener , Jian-Hua Zhang , Robert Heckel , Brian Herman
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