Article
Chemistry, Multidisciplinary
Nazar Oleksiievets, Christeena Mathew, Jan Christoph Thiele, Jose Ignacio Gallea, Oleksii Nevskyi, Ingo Gregor, Andre Weber, Roman Tsukanov, Joerg Enderlein
Summary: This study compares the performance of wide-field and confocal FL-SMLM techniques in different spectral regions and provides practical advice.
Article
Biochemical Research Methods
P. Wang, F. Hecht, G. Ossato, S. Tille, S. E. Fraser, J. A. Junge
Summary: Fluorescence lifetime imaging microscopy (FLIM) with phasor analysis allows for easy visualization and analysis of fluorophores' lifetimes, but typically suffers from low photon budgets leading to poor signal to noise ratios. Traditional median filters used in phasor analysis degrade high spatial frequency FLIM information, especially edges and puncta. A new filtering strategy using complex wavelet filtering and Anscombe transform preserves fine structures and accurate lifetimes in photon starved FLIM imaging, outperforming median filters and enabling FLIM imaging with lower laser power and faster imaging speed.
BIOMEDICAL OPTICS EXPRESS
(2021)
Article
Multidisciplinary Sciences
Cheng Zheng, Jong Kang Park, Murat Yildirim, Josiah R. Boivin, Yi Xue, Mriganka Sur, Peter T. C. So, Dushan N. Wadduwage
Summary: Nonlinear optical microscopy allows deep tissue imaging at millimeter scale in vivo, but has limited throughput. The DEEP technique combines patterned excitation and computational imaging to improve image quality in thick scattering samples, enabling reconstruction of high-resolution structural features from multiple DEEP images.
Article
Chemistry, Multidisciplinary
Adam J. Bowman, Mark A. Kasevich
Summary: This study demonstrates an electro-optic wide-field method using resonantly driven Pockels cells to achieve fluorescence lifetime microscopy (FLIM) with high throughput and single-molecule sensitivity. It allows capturing fluorescence lifetime of single molecules in wide field with fast dynamics and high sensitivity.
Article
Biochemical Research Methods
Rupsa Datta, Amani Gillette, Matthew Stefely, Melissa C. Skala
Summary: Fluorescence lifetime imaging microscopy (FLIM) is a powerful molecular imaging technique that measures the decay rate of fluorophores to provide insights into molecular interactions. Major advances in FLIM instrumentation, analysis, and biological and clinical applications over the last year have contributed to the growing interest in FLIM and continuous technological improvements in biomedical research.
JOURNAL OF BIOMEDICAL OPTICS
(2021)
Article
Biochemical Research Methods
Gilad Yahav, Shweta Pawar, Yitzchak Weber, Bar Atuar, Hamootal Duadi, Dror Fixler
Summary: Wide-field measurements of time-resolved fluorescence anisotropy (TR-FA) provide valuable information about the rotational mobility of fluorophores and have potential applications in cellular imaging and biochemical sensing. This study extends the existing fluorescence lifetime imaging microscopy (FLIM) to include TR-FA imaging (TR-FAIM), and applies it to the study of CD-gold nano conjugates. The results show significant changes in FLT, r, and. due to the attachment of gold nanoparticles to CDs.
JOURNAL OF BIOMEDICAL OPTICS
(2023)
Article
Engineering, Environmental
Adrian Monteleone, Folker Wenzel, Heinz Langhals, Daniel Dietrich
Summary: The FLIM method was tested for identification and characterization of six types of plastics, showing significant differentiation in 94.55% of the comparisons. Results suggest that FLIM has the potential for sub-micrometer plastic characterization and allows for visual 3D-sectioning of samples, which could be important for identifying and characterizing plastics in tissue and environmental samples.
JOURNAL OF ENVIRONMENTAL CHEMICAL ENGINEERING
(2021)
Article
Optics
Yide Zhang, Ian H. Guldner, Evan L. Nichols, David Benirschke, Cody J. Smith, Siyuan Zhang, Scott S. Howard
Summary: Instant FLIM is a method of analog signal processing that allows real-time streaming of fluorescence intensity, lifetime, and phasor imaging data, improving the efficiency and real-time capabilities of FLIM techniques. By upgrading existing two-photon microscopes with cost-effective components and open-source software, this technology can be easily implemented, along with enhancements in functionality through phasor segmentation, adaptive optics, and super-resolution techniques.
Article
Biochemical Research Methods
Yuyao Hu, Dong Liang, Jing Wang, Yaping Xuan, Fu Zhao, Jun Liu, Ruxin Li
Summary: The V-HiLo-ED method, based on edge detection, is proposed to remove background noise in fluorescence microscopy imaging. Compared with other software algorithms and hardware methods, V-HiLo-ED performs better in background removal and imaging quality, while also improving the signal-to-noise ratio of images.
JOURNAL OF BIOPHOTONICS
(2022)
Article
Biochemistry & Molecular Biology
Andrew H. A. Clayton
Summary: The dynamics of condensed matter can be studied using the time-dependent Stokes shift of a fluorescent probe. The time-dependent spectral correlation function is described by spectral relaxation correlation times that characterize the relaxation processes around the excited-state probe. Phasor plot is a powerful tool to analyze time and frequency-domain data, providing spatial maps of spectral dynamics in complex structures like living cells. The analysis workflow also addresses the extraction of initial and relaxed generalized polarization and improved methods for model discrimination and parameter extraction in complex spectral relaxation phenomena.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Optics
Alberto Ghezzi, Andrea Farina, Andrea Bassi, Gianluca Valentini, Ivan Labanca, Giulia Acconcia, Ivan Rech, Cosimo D'Andrea
Summary: Multispectral/hyperspectral fluorescence lifetime imaging microscopy (lambda FLIM) is a promising tool for studying biological processes. The development of a wide-field lambda FLIM setup with a novel single-photon avalanche diode array detector reduces measurement time by parallelizing spectral detection. The proposed system, incorporating a single-pixel camera with a compressive sensing scheme, represents an optimal microscopy framework for lambda FLIM setups.
Article
Multidisciplinary Sciences
Alice Sandmeyer, Lili Wang, Wolfgang Huebner, Marcel Mueller, Benjamin K. Chen, Thomas Huser
Summary: Cost-effective, highly portable, and easy to use high-resolution live-cell imaging systems can revolutionize research in challenging environments, enabling real-time tracking of virus particles and studying infection mechanisms.
Article
Chemistry, Multidisciplinary
Young Choi, MinKwan Kim, ChungHyun Park, Jongchan Park, YongKeun Park, Yong-Hoon Cho
Summary: This study presents a surface-sensitive super-resolution technique utilizing dynamic near-field speckle illumination, which enables the full super-resolving power of SOFI. Computational and experimental results demonstrate significant enhancements in spatial resolution, with the potential to achieve sub-100 nm resolution.
Article
Biochemical Research Methods
Miriam C. Bassler, Tim Rammler, Frank Wackenhut, Sven Zur Oven-Krockhaus, Ivona Secic, Rainer Ritz, Alfred J. Meixner, Marc Brecht
Summary: This study investigates the distribution and penetration behavior of hypericin in glioma cell spheroids using fluorescence microscopy and FLIM. The results show that hypericin exhibits a gradient distribution towards the spheroid core for short incubation periods or small concentrations, while a homogeneous distribution is observed for long incubation times or high concentrations.
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2022)
Article
Optics
Vahid Ebrahimi, Jialei Tang, Kyu Young Han
Summary: In this study, single-shot nondiffracting light-sheet microscopy was demonstrated through the incoherent superposition of dispersed polychromatic light sources. The technique was characterized by generating a Bessel light-sheet with a supercontinuum light-source and a C-light-sheet using a diode laser, showing its applicability in fluorescence microscopy. The method was emphasized to be easily implementable and compatible with the requirements of high-resolution microscopy.
Article
Optics
Saroch Leedumrongwatthanakun, Luca Innocenti, Hugo Defienne, Thomas Juffmann, Alessandro Ferraro, Mauro Paternostro, Sylvain Gigan
Article
Optics
Thomas Juffmann, Andres de los Rios Sommer, Sylvain Gigan
OPTICS COMMUNICATIONS
(2020)
Article
Microscopy
Stewart A. Koppell, Marian Mankos, Adam J. Bowman, Yonatan Israel, Thomas Juffmann, Brannon B. Klopfer, Mark A. Kasevich
Article
Physics, Applied
Dorian Bouchet, Jonathan Dong, Dante Maestre, Thomas Juffmann
Summary: A wide variety of imaging systems have been designed for measuring phase variations, with applications ranging from physics to biology and medicine. This work theoretically compares the precision of phase estimations achievable with classical phase microscopy techniques operating at the shot-noise limit. The study demonstrates the application of a general framework for the design and optimization of classical phase microscopes, showcasing the necessity of wavefront shaping for achieving optimal phase precision.
PHYSICAL REVIEW APPLIED
(2021)
Article
Instruments & Instrumentation
Brannon B. Klopfer, Stewart A. Koppell, Adam J. Bowman, Yonatan Israel, Mark A. Kasevich
Summary: The article presents the design and prototype of a switchable electron mirror, as well as a driving technique that pre-compensates pulses using an arbitrary waveform generator to achieve high fidelity waveform reproduction. The results demonstrate the feasibility of improving waveform accuracy in nonideal conditions using this method.
REVIEW OF SCIENTIFIC INSTRUMENTS
(2021)
Article
Chemistry, Multidisciplinary
Adam J. Bowman, Mark A. Kasevich
Summary: This study demonstrates an electro-optic wide-field method using resonantly driven Pockels cells to achieve fluorescence lifetime microscopy (FLIM) with high throughput and single-molecule sensitivity. It allows capturing fluorescence lifetime of single molecules in wide field with fast dynamics and high sensitivity.
Article
Physics, Applied
Jonathan Dong, Dante Maestre, Clara Conrad-Billroth, Thomas Juffmann
Summary: Interferometric imaging is a promising technique for particle tracking and mass photometry, allowing for precise measurement of parameters from weak signals coherently scattered from nanoparticles or single molecules. By computing the classical Cramer-Rao bound and quantum Cramer-Rao formalism, fundamental bounds on measurement precision can be derived, enabling comparison of different imaging techniques. The study demonstrates the increased axial position sensitivity in iSCAT and the minimum relative estimation error for mass estimation based on Quantum CRB.
JOURNAL OF PHYSICS D-APPLIED PHYSICS
(2021)
Article
Physics, Applied
Raphael Marchand, Radek Sachl, Martin Kalbac, Martin Hof, Rudolf Tromp, Mariana Amaro, Sense J. van der Molen, Thomas Juffmann
Summary: ONEM is an imaging technique that combines noninvasive probing with light and high-spatial-resolution readout via electron optics, providing label-free nanometric resolution without damaging the specimen. By converting optical near fields into a spatially varying electron flux and using low-energy electron microscopy, ONEM enables imaging without the need to scan a probe across the sample.
PHYSICAL REVIEW APPLIED
(2021)
Article
Art
Enar De Dios Rodriguez, Brannon B. Klopfer, Philipp Haslinger, Thomas Juffmann
Summary: SEEC photography is a project that blends art and science, utilizing modern technology to record the movement of light and drawing inspiration from the universal physical constant for the speed of light. The project aims to familiarize the general public with this physical phenomenon by capturing light moving across familiar objects, paying homage to iconic images from the history of photography. By using exposure times shorter than 0.3 nanoseconds, the authors are able to capture light in the process of writing an image.
Article
Physics, Applied
Stewart A. Koppell, Yonatan Israel, Adam J. Bowman, Brannon B. Klopfer, M. A. Kasevich
Summary: The article discusses a framework for calculating the amount of information carried by each electron, evaluating the potential effectiveness of different microscopes, and points out that the quantum limit in phase imaging is significantly higher than currently achievable levels.
APPLIED PHYSICS LETTERS
(2022)
Article
Physics, Multidisciplinary
Marius Constantin Chirita Mihaila, Philipp Weber, Matthias Schneller, Lucas Grandits, Stefan Nimmrichter, Thomas Juffmann
Summary: This study demonstrates for the first time programmable transverse electron-beam shaping in free space based on ponderomotive potentials from short intense laser pulses. Both convex and concave electron lenses with a similar focal length to state-of-the-art electron microscopes can be realized, and almost arbitrary deflection patterns can be achieved by shaping the ponderomotive potentials using a spatial light modulator.
Article
Physics, Applied
Joshua L. Reynolds, Yonatan Israel, Adam J. Bowman, Brannon B. Klopfer, Mark A. Kasevich
Summary: In this study, photoemission from a Schottky emitter triggered by nanosecond laser pulses is demonstrated. By using photon energies optimally tuned to the emission potential barrier, pulses containing over 105 electrons with energy spreads below 1 eV are generated through a single-photon photoemission process. These results can have widespread implementation and are consistent with a theoretical model of laser-triggered electron emission and energetic broadening during propagation.
PHYSICAL REVIEW APPLIED
(2023)
Article
Physics, Applied
Yonatan Israel, Adam J. Bowman, Brannon B. Klopfer, Stewart A. Koppell, Mark A. Kasevich
APPLIED PHYSICS LETTERS
(2020)
Article
Optics
Stefan Nimmrichter, Chi-Fang Chen, Brannon B. Klopfer, Mark A. Kasevich, Thomas Juffmann
JOURNAL OF PHYSICS-PHOTONICS
(2019)