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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

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

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

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

The investigation of fluorescence lifetime became an important tool in biology and medical science. So far, established methods of fluorescence lifetime measurements require the illumination of the investigated probes with pulsed or…

Fluorescence Lifetime Imaging Microscopy (FLIM) using multiphoton excitation techniques is now finding an important place in quantitative imaging of protein-protein interactions and intracellular physiology. We review here the recent…

Quantitative Methods · Quantitative Biology 2009-11-10 R. V. Krishnan , A. Masuda , V. E. Centoze , B. Herman

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

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

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…

Fluorescence lifetime imaging microscopy (FLIM) provides detailed information about molecular interactions and biological processes. A major bottleneck for FLIM is image resolution at high acquisition speeds, due to the engineering and…

Image and Video Processing · Electrical Eng. & Systems 2024-04-23 Valentin Kapitány , Areeba Fatima , Vytautas Zickus , Jamie Whitelaw , Ewan McGhee , Robert Insall , Laura Machesky , Daniele Faccio

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

We present a fast and accurate analytical method for fluorescence lifetime imaging microscopy (FLIM) using the extreme learning machine (ELM). We used extensive metrics to evaluate ELM and existing algorithms. First, we compared these…

Biological Physics · Physics 2022-03-28 Zhenya Zang , Dong Xiao , Quan Wang , Zinuo Li , Wujun Xie , Yu Chen , David Day Uei Li

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

Despite their widespread use in cell biology, fluorescence lifetime imaging microscopy (FLIM) data-sets are challenging to analyse, because each spatial position can contain a superposition of multiple fluorescent components. Here, we…

Biological Physics · Physics 2022-08-29 Francesco Masia , Walter Dewitte , Paola Borri , Wolfgang Langbein

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

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

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…

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…

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…

Interaction-free measurement (IFM) is a promising technique for low-dose detection and imaging, offering the unique advantage of probing an object without absorption of the interrogating photons. We propose an experiment to demonstrate IFM…

Quantum Physics · Physics 2024-07-10 Ron Cohen , Sharon Shwartz , Eliahu Cohen
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