Two‐Photon Excitation Microscopy for the Study of Living Cells and Tissues

Benninger, Richard K.P.; Piston, David W.

doi:10.1002/0471143030.cb0411s59

Cited by 208 publications

(169 citation statements)

References 84 publications

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“…Although Ca 2þ and NAD(P)H imaging can illuminate the complex intracellular mechanisms in association with insulin secretion, these imaging techniques require the use of confocal microscopy or two photon microscopy. 16 The method of islet immobilization on different microfluidic platforms varies, including use of mechanical actuators 15 and well arrays. 9,17 Using actuators requires complicated islet loading procedures, prevents good perifusion around the islets, and subjects islets to mechanical stress.…”

Section: Introductionmentioning

confidence: 99%

Microfluidic platform for assessing pancreatic islet functionality through dielectric spectroscopy

et al. 2015

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Human pancreatic islets are seldom assessed for dynamic responses to external stimuli. Thus, the elucidation of human islet functionality would provide insights into the progression of diabetes mellitus, evaluation of preparations for clinical transplantation, as well as for the development of novel therapeutics. The objective of this study was to develop a microfluidic platform for in vitro islet culture, allowing the multi-parametric investigation of islet response to chemical and biochemical stimuli. This was accomplished through the fabrication and implementation of a microfluidic platform that allowed the perifusion of islet culture while integrating real-time monitoring using impedance spectroscopy, through microfabricated, interdigitated electrodes located along the microchamber arrays. Real-time impedance measurements provide important dielectric parameters, such as cell membrane capacitance and cytoplasmic conductivity, representing proliferation, differentiation, viability, and functionality. The perifusion of varying glucose concentrations and monitoring of the resulting impedance of pancreatic islets were performed as proof-of-concept validation of the lab-on-chip platform. This novel technique to elucidate the underlying mechanisms that dictate islet functionality is presented, providing new information regarding islet function that could improve the evaluation of islet preparations for transplantation. In addition, it will lead to a better understanding of fundamental diabetes-related islet dysfunction and the development of therapeutics through evaluation of potential drug effects. V C 2015 AIP Publishing LLC. [http://dx

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Section: Introductionmentioning

confidence: 99%

Microfluidic platform for assessing pancreatic islet functionality through dielectric spectroscopy

et al. 2015

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“…Emissions were detected by nondescanned detectors [3], the emitted light being split between blue (<485 nm), green (500-620 nm) and red (>620 nm) channels for simultaneous recording. A detector was also available for transmitted light observations.…”

Section: Multiphoton Confocal Imagingmentioning

confidence: 99%

Multicolour correlative imaging using phosphor probes

et al. 2015

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“…by Kaiser and Garret when exciting an europium-doped crystal [15]. To date, two-photon fluorescence is mostly used in biomedical engineering, when studying living cells and tissue by using two-photon fluorescence microscopy [16][17][18]. Intuitively, we would expect that all fluorescent molecules, able to emit OPIF, generate a TPIF signal if they are excited with laser light with the double wavelength and a sufficiently high excitation power density.…”

Section: Introductionmentioning

confidence: 99%

Improving Food Safety by Using One- and Two-Photon- Induced Fluorescence Spectroscopy for the Detection of Mycotoxins

Smeesters¹,

Meulebroeck²,

Thienpont³

2016

Applications of Molecular Spectroscopy to Current Research in the Chemical and Biological Sciences

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The presence of mycotoxins in food products is a major worldwide problem. Nowadays, mycotoxins can only be detected by the use of sample-based chemical analyses. Therefore, we demonstrate the use of one-and two-photon-induced fluorescence spectroscopy for the non-destructive detection of mycotoxins in unprocessed food products. We first explain our optical set-up, which is able to measure the localized oneand two-photon-induced fluorescence spectra. Following, as a case study, the detection of aflatoxin in maize kernels is discussed. We present our research methodology, from the characterization of the fluorescence of pure aflatoxin, to the study of the one-and two-photon-induced fluorescence spectra of maize kernels and the development of an optical detection criterion. During both one-and two-photon-induced fluorescence processes, the fluorescence of the aflatoxin influences the intrinsic fluorescence of the maize. Based on the fluorescence spectrum between 400 and 550 nm, a detection criterion to sense the contaminated kernels is defined. Furthermore, we successfully monitored the localized contamination level on the kernel's surface, showing both contaminated kernels with a high contamination in a limited surface area (a few square millimetres) and kernels with a low contamination spread over a large surface area (up to 20 mm 2 ). Finally, the extensibility of our research methodology to other fluorescent mycotoxins is discussed.

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Two‐Photon Excitation Microscopy for the Study of Living Cells and Tissues

Cited by 208 publications

References 84 publications

Microfluidic platform for assessing pancreatic islet functionality through dielectric spectroscopy

Microfluidic platform for assessing pancreatic islet functionality through dielectric spectroscopy

Multicolour correlative imaging using phosphor probes

Improving Food Safety by Using One- and Two-Photon- Induced Fluorescence Spectroscopy for the Detection of Mycotoxins

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