A model for correcting the fluorescence signal from a free-falling depth profiler

2006 ◽  
Vol 40 (8) ◽  
pp. 1616-1626 ◽  
Author(s):  
Jens Schimanski ◽  
Martin Beutler ◽  
Christian Moldaenke ◽  
Ulf-Peter-Hansen
Author(s):  
Stephen R. Bolsover

The field of intracellular ion concentration measurement expanded greatly in the 1980's due primarily to the development by Roger Tsien of ratiometric fluorescence dyes. These dyes have many applications, and in particular they make possible to image ion concentrations: to produce maps of the ion concentration within living cells. Ion imagers comprise a fluorescence microscope, an imaging light detector such as a video camera, and a computer system to process the fluorescence signal and display the map of ion concentration.Ion imaging can be used for two distinct purposes. In the first, the imager looks at a field of cells, measuring the mean ion concentration in each cell of the many in the field of view. One can then, for instance, challenge the cells with an agonist and examine the response of each individual cell. Ion imagers are not necessary for this sort of experiment: one can instead use a system that measures the mean ion concentration in a just one cell at any one time. However, they are very much more convenient.


2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiang Lan Fan ◽  
Jose A. Rivera ◽  
Wei Sun ◽  
John Peterson ◽  
Henry Haeberle ◽  
...  

AbstractUnderstanding the structure and function of vasculature in the brain requires us to monitor distributed hemodynamics at high spatial and temporal resolution in three-dimensional (3D) volumes in vivo. Currently, a volumetric vasculature imaging method with sub-capillary spatial resolution and blood flow-resolving speed is lacking. Here, using two-photon laser scanning microscopy (TPLSM) with an axially extended Bessel focus, we capture volumetric hemodynamics in the awake mouse brain at a spatiotemporal resolution sufficient for measuring capillary size and blood flow. With Bessel TPLSM, the fluorescence signal of a vessel becomes proportional to its size, which enables convenient intensity-based analysis of vessel dilation and constriction dynamics in large volumes. We observe entrainment of vasodilation and vasoconstriction with pupil diameter and measure 3D blood flow at 99 volumes/second. Demonstrating high-throughput monitoring of hemodynamics in the awake brain, we expect Bessel TPLSM to make broad impacts on neurovasculature research.


Molecules ◽  
2020 ◽  
Vol 25 (4) ◽  
pp. 781
Author(s):  
Ernesto Enríquez-Palacios ◽  
Teresa Arbeloa ◽  
Jorge Bañuelos ◽  
Claudia I. Bautista-Hernández ◽  
José G. Becerra-González ◽  
...  

Herein we report on a straightforward access method for boron dipyrromethene dyes (BODIPYs)-coumarin hybrids linked through their respective 8- and 6- positions, with wide functionalization of the coumarin fragment, using salicylaldehyde as a versatile building block. The computationally-assisted photophysical study unveils broadband absorption upon proper functionalization of the coumarin, as well as the key role of the conformational freedom of the coumarin appended at the meso position of the BODIPY. Such free motion almost suppresses the fluorescence signal, but enables us to apply these dyads as molecular rotors to monitor the surrounding microviscosity.


2020 ◽  
Vol 74 (4) ◽  
pp. 439-451
Author(s):  
Philipp Holz ◽  
Christoph Pönisch ◽  
Albrecht Brandenburg

Imaging fluorescence spectroscopy proves to be a fast and sensitive method for measuring the thickness of thin coatings in the manufacturing industry. This encouraged us to systematically study, theoretically and experimentally, parameters that influence the fluorescence of thin layers. We analyzed the fluorescence signal as a function of the scattering and reflectance properties of the sample substrate. In addition, we investigated effects of the layer properties on fluorescence emission. A ray-tracing software is used to describe the influence of these parameters on the fluorescence emission of thin layers. Experiments using a custom-made system for imaging fluorescence analysis verify the simulations. This work shows a factor five variation of fluorescence intensity as a function of the reflectance of the sample substrate. Simulations show variations by a factor of up to eight for samples with different surface roughness. Results on tilted samples indicate a significant increase of the detected fluorescence signal, for fluorescent droplets on reflective substrates, if illuminated and coaxially observed at angles greater than 25°. These findings are of utmost relevance for all applications which utilize the fluorescence emission to quantify thin layers. These applications range from in-line lubricant monitoring in press plants to monitoring of functional coatings in medical technology and the detection of filmic contaminations.


2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Yipengchen Yin ◽  
Yongjing Li ◽  
Sheng Wang ◽  
Ziliang Dong ◽  
Chao Liang ◽  
...  

Abstract Background The recently developed biomimetic strategy is one of the mostly effective strategies for improving the theranostic efficacy of diverse nanomedicines, because nanoparticles coated with cell membranes can disguise as “self”, evade the surveillance of the immune system, and accumulate to the tumor sites actively. Results Herein, we utilized mesenchymal stem cell memabranes (MSCs) to coat polymethacrylic acid (PMAA) nanoparticles loaded with Fe(III) and cypate—an derivative of indocyanine green to fabricate Cyp-PMAA-Fe@MSCs, which featured high stability, desirable tumor-accumulation and intriguing photothermal conversion efficiency both in vitro and in vivo for the treatment of lung cancer. After intravenous administration of Cyp-PMAA-Fe@MSCs and Cyp-PMAA-Fe@RBCs (RBCs, red blood cell membranes) separately into tumor-bearing mice, the fluorescence signal in the MSCs group was 21% stronger than that in the RBCs group at the tumor sites in an in vivo fluorescence imaging system. Correspondingly, the T1-weighted magnetic resonance imaging (MRI) signal at the tumor site decreased 30% after intravenous injection of Cyp-PMAA-Fe@MSCs. Importantly, the constructed Cyp-PMAA-Fe@MSCs exhibited strong photothermal hyperthermia effect both in vitro and in vivo when exposed to 808 nm laser irradiation, thus it could be used for photothermal therapy. Furthermore, tumors on mice treated with phototermal therapy and radiotherapy shrank 32% more than those treated with only radiotherapy. Conclusions These results proved that Cyp-PMAA-Fe@MSCs could realize fluorescence/MRI bimodal imaging, while be used in phototermal-therapy-enhanced radiotherapy, providing desirable nanoplatforms for tumor diagnosis and precise treatment of non-small cell lung cancer.


BJS Open ◽  
2021 ◽  
Vol 5 (2) ◽  
Author(s):  
M D Slooter ◽  
M S E Mansvelders ◽  
P R Bloemen ◽  
S S Gisbertz ◽  
W A Bemelman ◽  
...  

Abstract Background The aim of this systematic review was to identify all methods to quantify intraoperative fluorescence angiography (FA) of the gastrointestinal anastomosis, and to find potential thresholds to predict patient outcomes, including anastomotic leakage and necrosis. Methods This systematic review adhered to the PRISMA guidelines. A PubMed and Embase literature search was performed. Articles were included when FA with indocyanine green was performed to assess gastrointestinal perfusion in human or animals, and the fluorescence signal was analysed using quantitative parameters. A parameter was defined as quantitative when a diagnostic numeral threshold for patient outcomes could potentially be produced. Results Some 1317 articles were identified, of which 23 were included. Fourteen studies were done in patients and nine in animals. Eight studies applied FA during upper and 15 during lower gastrointestinal surgery. The quantitative parameters were divided into four categories: time to fluorescence (20 studies); contrast-to-background ratio (3); pixel intensity (2); and numeric classification score (2). The first category was subdivided into manually assessed time (7 studies) and software-derived fluorescence–time curves (13). Cut-off values were derived for manually assessed time (speed in gastric conduit wall) and derivatives of the fluorescence–time curves (Fmax, T1/2, TR and slope) to predict patient outcomes. Conclusion Time to fluorescence seems the most promising category for quantitation of FA. Future research might focus on fluorescence–time curves, as many different parameters can be derived and the fluorescence intensity can be bypassed. However, consensus on study set-up, calibration of fluorescence imaging systems, and validation of software programs is mandatory to allow future data comparison.


Sensors ◽  
2021 ◽  
Vol 21 (4) ◽  
pp. 1041
Author(s):  
Mazhar Hussain ◽  
Mattias O’Nils ◽  
Jan Lundgren

High temperatures complicate the direct measurements needed for continuous characterization of the properties of molten materials such as glass. However, the assumption that geometrical changes when the molten material is in free-fall can be correlated with material characteristics such as viscosity opens the door to a highly accurate contactless method characterizing small dynamic changes. This paper proposes multi-camera setup to achieve accuracy close to the segmentation error associated with the resolution of the images. The experimental setup presented shows that the geometrical parameters can be characterized dynamically through the whole free-fall process at a frame rate of 600 frames per second. The results achieved show the proposed multi-camera setup is suitable for estimating the length of free-falling molten objects.


2021 ◽  
Author(s):  
Cameron R. S. Thompson ◽  
James E. Bron ◽  
Samantha Bui ◽  
Sussie Dalvin ◽  
Mark J. Fordyce ◽  
...  

2021 ◽  
Vol 22 (15) ◽  
pp. 8256
Author(s):  
Adolfas K. Gaigalas ◽  
Yu-Zhong Zhang ◽  
Linhua Tian ◽  
Lili Wang

A stochastic model of the flow cytometer measurement process was developed to assess the nature of the observed coefficient of variation (CV%) of the mean fluorescence intensity (MFI) from a population of labeled microspheres (beads). Several sources of variability were considered: the total number of labels on a bead, the path through the laser beam, the optical absorption cross-section, the quantum yield, the numerical aperture of the collection optics, and the photoelectron conversion efficiency of the photomultiplier (PMT) cathode. The variation in the number of labels on a bead had the largest effect on the CV% of the MFI of the bead population. The variation in the path of the bead through the laser beam was minimized using flat-top lasers. The variability in the average optical properties of the labels was of minor importance for beads with sufficiently large number of labels. The application of the bead results to the measured CV% of labeled B cells indicated that the measured CV% was a reliable measure of the variability of antibodies bound per cell. With some modifications, the model can be extended to multicolor flow cytometers and to the study of CV% from cells with low fluorescence signal.


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