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Nanosecond temporal resolution enables new methods for wide-field imaging like time-of-flight, gated detection, and fluorescence lifetime. The optical efficiency of existing approaches, however, presents challenges for low-light…

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…

The development of voltage-sensitive fluorescent probes suggests fluorescence lifetime as a promising readout for electrical activity in biological systems. Existing approaches fail to achieve the speed and sensitivity required for voltage…

Optics · Physics 2023-06-27 Adam J. Bowman , Cheng Huang , Mark J. Schnitzer , Mark A. Kasevich

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…

Photoluminesce (PL) spectroscopy offers excellent methods for mapping the PL decay on the nanosecond time scale. However, capturing maps of emission dynamics on the microsecond time scale can be highly time-consuming. We present a new…

Optics · Physics 2021-02-24 Jiri Junek , Karel Zidek

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…

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

Two-Photon Microscopy has become an invaluable tool for biological and medical research, providing high sensitivity, molecular specificity, inherent three-dimensional sub-cellular resolution and deep tissue penetration. In terms of imaging…

Fluorescence lifetime imaging microscopy (FLIM) is a powerful tool for quantitative fluorescence imaging because fluorescence lifetime is independent of concentration of fluorescent molecules or excitation/detection efficiency and is robust…

Endoscopic optical imaging using a single multimode fiber (MMF) has emerged as a promising approach for highly compact, minimally invasive, and high-resolution imaging. Unlike conventional fiber bundles, MMF-based endomicroscopes exploit…

Two-photon excited fluorescence (TPEF) microscopy and fluorescence lifetime imaging (FLIM) are powerful imaging techniques in bio-molecular science. The need for elaborate light sources for TPEF and speed limitations for FLIM, however,…

Instrumentation and Detectors · Physics 2018-10-12 Matthias Eibl , Sebastian Karpf , Hubertus Hakert , Daniel Weng , Robert Huber

Fluorescence lifetime imaging microscopy (FLIM) is an important technique to understand the chemical micro-environment in cells and tissues since it provides additional contrast compared to conventional fluorescence imaging. When two…

Image and Video Processing · Electrical Eng. & Systems 2022-01-19 Varun Mannam , Xiaotong Yuan , Scott Howard

Fluorescence lifetime imaging microscopy (FLIM) is a powerful quantitative technique that provides metabolic and molecular contrast, offering strong translational potential for label-free, real-time diagnostics. However, its clinical…

Computer Vision and Pattern Recognition · Computer Science 2025-12-19 Paloma Casteleiro Costa , Parnian Ghapandar Kashani , Xuhui Liu , Alexander Chen , Ary Portes , Julien Bec , Laura Marcu , Aydogan Ozcan

Fluorescence lifetime imaging microscopy (FLIM) is a well-established technique with numerous imaging applications. Yet, one of the limitations of FLIM is that it provides information about the emitting state only. Here, we present an…

Biological Physics · Physics 2023-10-27 Pavel Malý , Dita Strachotová , Aleš Holoubek , Petr Heřman

RAndom Temporal Signals (RATS) method has proven to be a useful and versatile method for measuring photoluminescence (PL) dynamics and fluorescence lifetime imaging (FLIM). Here, we present two fundamental development steps in the method.…

Optics · Physics 2023-02-15 Jiri Junek , Karel Zidek

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

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

Microscopy has been key to tremendous advances in science, technology, and medicine, revealing structure and dynamics across time and length scales. However, combining high spatial and temporal resolution in a non-invasive, label-free…

Fluorescence lifetime imaging microscopy (FLIM) is a powerful technique in biomedical research that uses the fluorophore decay rate to provide additional contrast in fluorescence microscopy. However, at present, the calculation, analysis,…

Image and Video Processing · Electrical Eng. & Systems 2021-06-09 Varun Mannam , Yide Zhang , Xiaotong Yuan , Cara Ravasio , Scott S. Howard

FRET measurements can provide dynamic spatial information on length scales smaller than the diffraction limit of light. Several methods exist to measure FRET between fluorophores, including Fluorescence Lifetime Imaging Microscopy (FLIM),…

Quantitative Methods · Quantitative Biology 2017-01-09 Bryan Kaye , Peter J. Foster , Tae Yeon Yoo , Daniel J. Needleman
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