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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) 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

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

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

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 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

Luminescence imaging is invaluable for studying biological and material systems, particularly when advanced protocols that exploit temporal dynamics are employed. However, implementing such protocols often requires custom instrumentation,…

Instrumentation and Detectors · Physics 2025-09-16 Ian Coghill , Alienor Lahlou , Andrea Lodetti , Shizue Matsubara , Johann Boucle , Thomas Le Saux , Ludovic Jullien

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) 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

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…

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

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

Fluorescence spectroscopy is an image correlation technique to analyze and characterize the molecular dynamics from a sequence of fluorescence images. Many image correlation techniques have been developed for different applications [1]. But…

Quantitative Methods · Quantitative Biology 2017-10-24 C. Chen , P. Paul-Gilloteaux , T. Vignaud , J. Salamero , F. Waharte

The evolution, regulation and sustenance of biological complexity is determined by protein-protein interaction network that is filled with dynamic events. Recent experimental evidences point out that clustering of proteins has a vital role…

Tissues and Organs · Quantitative Biology 2007-05-23 RV Krishnan

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

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

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

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…

Temporally and spatially resolved measurements of protein transport inside cells provide important clues to the functional architecture and dynamics of biological systems. Fluorescence Recovery After Photobleaching (FRAP) technique has been…

Subcellular Processes · Quantitative Biology 2009-03-04 George D. Tsibidis
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