We describe the performance of a new wide area time-gated single-photon avalanche diode (SPAD) array for phasor-FLIM, exploring the effect of gate length, gate number and signal intensity on the measured lifetime accuracy and precision. We conclude that the detector functions essentially as an ideal shot noise limited sensor and is capable of video rate FLIM measurement. The phasor approach used in this work appears ideally suited to handle the large amount of data generated by this type of very large sensor (512 × 512 pixels), even in the case of small number of gates and limited photon budget.
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http://dx.doi.org/10.1088/2050-6120/ab6ed7 | DOI Listing |
Spectrochim Acta A Mol Biomol Spectrosc
October 2024
LaserLaB, Faculty of Sciences, Vrije Universiteit Amsterdam, de Boelelaan 1081, 1081 HV, Amsterdam, The Netherlands.
Raman spectroscopy is a well-established method for chemical identification, with a wide variety of applications. The two major limitations are that fluorescence can hamper detection, and that Raman imaging is slow; it typically takes multiple hours to measure even a small surface area. We have developed a multimodal setup that mitigates these limitations.
View Article and Find Full Text PDFJ Biomed Opt
February 2024
Ariel University, Department of Physics, Faculty of Natural Science, Ariel, Israel.
Significance: Fluorescence lifetime imaging (FLI) plays a pivotal role in enhancing our understanding of biological systems, providing a valuable tool for non-invasive exploration of biomolecular and cellular dynamics, both and . Its ability to selectively target and multiplex various entities, alongside heightened sensitivity and specificity, offers rapid and cost-effective insights.
Aim: Our aim is to investigate the multiplexing capabilities of near-infrared (NIR) FLI within a scattering medium that mimics biological tissues.
Discov Nano
January 2024
Department of Physics, Faculty of Natural Science, Ariel University, 40700, Ariel, Israel.
The near-infrared (NIR) range of the electromagnetic (EM) spectrum offers a nearly transparent window for imaging tissue. Despite the significant potential of NIR fluorescence-based imaging, its establishment in basic research and clinical applications remains limited due to the scarcity of fluorescent molecules with absorption and emission properties in the NIR region, especially those suitable for biological applications. In this study, we present a novel approach by combining the widely used IRdye 800NHS fluorophore with gold nanospheres (GNSs) and gold nanorods (GNRs) to create Au nanodyes, with improved quantum yield (QY) and distinct lifetimes.
View Article and Find Full Text PDFIn biological research, rapid wide-field fluorescence lifetime imaging has become an important imaging tool. However, the biological samples with weak fluorescence signals and lower sensitivity often suffer from very low precision in lifetime determinations which restricts its widespread utilization in many bioimaging applications. To address this issue, a method is presented in this paper to substantially enhance the precision of rapid lifetime determination (RLD).
View Article and Find Full Text PDFJ Biomed Opt
October 2022
Univ. of Wisconsin-Madison, United States.
Significance: Hypoxia imaging for surgical guidance has never been possible, yet it is well known that most tumors have microregional chronic and/or cycling hypoxia present as well as chaotic blood flow. The ability to image oxygen partial pressure (pO2) is therefore a unique control of tissue metabolism and can be used in a range of disease applications to understand the complex biochemistry of oxygen supply and consumption.
Aim: Delayed fluorescence (DF) from the endogenous molecule protoporphyrin IX (PpIX) has been shown to be a truly unique reporter of the local oxygen partial pressure in tissue.
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