scholarly journals Multicolor fluorescent imaging by space-constrained computational hyperspectral imaging

2018 ◽  
Author(s):  
Yina Wang ◽  
Bin Yang ◽  
Siyu Feng ◽  
Veronica Pessino ◽  
Bo Huang
Keyword(s):  
Experimental Data ◽  
Hyperspectral Imaging ◽  
Fluorescent Proteins ◽  
Emission Spectra ◽  
Fluorescent Imaging ◽  
Cellular Structures ◽  
Imaging Method ◽  
Color Imaging ◽  
Powerful Technique ◽  
Under Sampling

AbstractHyperspectral imaging is a powerful technique to simultaneously study multiple fluorophore labels with overlapping emissions. Here we present a computational hyperspectral imaging method, which uses the sample spatial fluorescence information as a reconstruction constraint. Our method addresses both the under-sampling issue of compressive hyperspectral imaging and the low throughput issue of scanning hyperspectral imaging. With simulated and experimental data, we have demonstrated the superior reconstruction precision of our method in two and three-color imaging. We have experimentally validated this method in differentiating cellular structures labeled with two red-colored fluorescent proteins, tdTomato and mCherry, which have highly overlapping emission spectra. Our method has the advantage of totally free wavelength choice and can also be combined with conventional filter-based sequential multi-color imaging to further expand the choices of probes.

scholarly journals Hyperspectral imaging as a tool for assessing coral health utilising natural fluorescence

2019 ◽  
Author(s):  
Jonathan Teague ◽  
Jack Willans ◽  
Michael Allen ◽  
Thomas Scott ◽  
John Day
Keyword(s):  
Hyperspectral Imaging ◽  
Coral Bleaching ◽  
Cell Biology ◽  
Fluorescent Proteins ◽  
Emission Spectra ◽  
Pocillopora Damicornis ◽  
Remotely Operated Vehicle ◽  
Underwater Visual Census ◽  
Research Fields ◽  
Natural Fluorescence

Fluorescent proteins are a crucial visualisation tool in a myriad of research fields including cell biology, microbiology and medicine. Fluorescence is a result of the absorption of electromagnetic radiation at one wavelength and its reemission at a longer wavelength. Coral communities exhibit a natural fluorescence which can be used to distinguish between diseased and healthy specimens, however, current methods, such as the underwater visual census, are expensive and time-consuming constituting many manned dive hours. We propose the use of a remotely operated vehicle mounted with a novel hyperspectral fluorescence imaging (HyFI) “payload” for more rapid surveying and data collection. We have tested our system in a laboratory environment on common coral species including Seriatopora spp., Montipora verrucosa, Montipora spp., Montipora capricornis, Echinopora lamellose, Euphyllia ancora, Pocillopora damicornis and Montipora confusa. With the aid of hyperspectral imaging, the coral specimens’ emission wavelengths can be accurately assessed by capturing the emission spectra of the corals when excited with light emitting diodes (395–405 and 440 nm). Fluorescence can also provide an indicator of coral bleaching as shown in our bleaching experiment where we observe fluorescence reduction alongside coral bleaching.

Fluorescent proteins for in vivo imaging, where's the biliverdin?

2020 ◽  
Vol 48 (6) ◽  
pp. 2657-2667
Author(s):  
Felipe Montecinos-Franjola ◽  
John Y. Lin ◽  
Erik A. Rodriguez
Keyword(s):  
Hydrogen Peroxide ◽  
In Vivo Imaging ◽  
Near Infrared ◽  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Fluorescent Imaging ◽  
Infrared Light ◽  
Red Fluorescent Protein ◽  
Color Palette

Noninvasive fluorescent imaging requires far-red and near-infrared fluorescent proteins for deeper imaging. Near-infrared light penetrates biological tissue with blood vessels due to low absorbance, scattering, and reflection of light and has a greater signal-to-noise due to less autofluorescence. Far-red and near-infrared fluorescent proteins absorb light >600 nm to expand the color palette for imaging multiple biosensors and noninvasive in vivo imaging. The ideal fluorescent proteins are bright, photobleach minimally, express well in the desired cells, do not oligomerize, and generate or incorporate exogenous fluorophores efficiently. Coral-derived red fluorescent proteins require oxygen for fluorophore formation and release two hydrogen peroxide molecules. New fluorescent proteins based on phytochrome and phycobiliproteins use biliverdin IXα as fluorophores, do not require oxygen for maturation to image anaerobic organisms and tumor core, and do not generate hydrogen peroxide. The small Ultra-Red Fluorescent Protein (smURFP) was evolved from a cyanobacterial phycobiliprotein to covalently attach biliverdin as an exogenous fluorophore. The small Ultra-Red Fluorescent Protein is biophysically as bright as the enhanced green fluorescent protein, is exceptionally photostable, used for biosensor development, and visible in living mice. Novel applications of smURFP include in vitro protein diagnostics with attomolar (10−18 M) sensitivity, encapsulation in viral particles, and fluorescent protein nanoparticles. However, the availability of biliverdin limits the fluorescence of biliverdin-attaching fluorescent proteins; hence, extra biliverdin is needed to enhance brightness. New methods for improved biliverdin bioavailability are necessary to develop improved bright far-red and near-infrared fluorescent proteins for noninvasive imaging in vivo.

Labeling cellular structures in vivo using confined primed conversion of photoconvertible fluorescent proteins

2016 ◽  
Vol 11 (12) ◽  
pp. 2419-2431 ◽  
Author(s):  
Manuel Alexander Mohr ◽  
Paul Argast ◽  
Periklis Pantazis
Keyword(s):  
Fluorescent Proteins ◽  
Cellular Structures

Study on the equivalent luminance of luminous fibers in photopic vision and scotopic vision

2017 ◽  
Vol 88 (10) ◽  
pp. 1157-1163 ◽  
Author(s):  
Yanhong Yan ◽  
Chengxia Liu ◽  
Xiaojun Ding
Keyword(s):  
Experimental Data ◽  
Rare Earth ◽  
Melt Spinning ◽  
Emission Spectra ◽  
Scotopic Vision ◽  
Photopic Vision ◽  
Earth Material ◽  
Long Afterglow ◽  
Polyamide Fiber ◽  
Spinning Process

Colored luminous fibers were prepared by a melt spinning process, adding colored pigments and long afterglow rare earth material into polyamide fiber. The colored luminous fibers had a variety of colors in photopic vision, and emitted colored light in mesopic vision and scotopic vision. Based on the experimental data of the emission spectra and the test luminance of the luminous fibers, the effect of the emissive colors of the luminous fibers on the equivalent luminance at different vision states was analyzed. The results showed that the effect of the emissive colors of white, red, yellow, and green luminous fibers on the equivalent luminance was not obvious in photopic and scotopic vision, but that of blue luminous fiber was obvious in photopic vision but not in scotopic vision.

Measurement Tools for Non-Invasive Monitoring of the Plants Growth Conditions by Using Hyperspectral Imaging Methods: a Review

2021 ◽  
Vol 2 (43) ◽  
pp. 54-61
Author(s):  
Dmitriy A. Burynin ◽  
◽  
Aleksandr A. Smirnov
Keyword(s):  
Hyperspectral Imaging ◽  
Physiological State ◽  
Effective Means ◽  
Research Problem ◽  
High Spectral Resolution ◽  
Research Purpose ◽  
Imaging Method ◽  
Invasive Monitoring ◽  
Imaging Methods ◽  
Non Invasive

Portable spectroradiometers and hyperspectral cameras are increasingly being used to quickly assess the physiological state of plants. The operation of these devices is based on the registration of reflection or reflection and transmission spectra. (Research purpose) The research purpose is in analyzing the technical means and methods of non-invasive monitoring of the plant state based on the registration of the reflection spectra of leaves. (Materials and methods) The article presents a review of the work on the application of hyperspectral imaging methods. Authors classified and analyzed materials on spectroscopic radiometers and hyperspectral cameras, and outlined the prospects for implementation. Authors applied the methods of a systematic approach to the research problem. (Results and discussion) Hyperspectral imaging methods serve as an effective means of monitoring plants. It is possible to determine the pigment composition of plants, lack of nutrition, and detect biotic stress through hyperspectral imaging. The article presents methods of application of portable spectroradiometers and hyperspectral cameras. With the help of these devices it is possible to carry out measurements with high spectral resolution. The difficulty of accurately detecting the content of pigments in the leaves lies in the mutual overlap of the areas of light absorption by them. The main drawback of spectroradiometers is that they measure only at one point on a single sheet. The article presents the difficulties encountered in interpreting the results obtained by the hyperspectral camera. The background reflectivity of the soil, the geometry of the vegetation cover, and the uneven lighting can make errors in the measurements. (Conclusions) The article presents the disadvantages of the hyperspectral imaging method when using only the reflection spectrum. In order to increase the accuracy of the determination of pigments and stresses of various origins, it is necessary to develop a portable device that combines the methods of recording reflection and fluorescence.

scholarly journals Comparative Analysis of Bacteriophytochrome Agp2 and Its Engineered Photoactivatable NIR Fluorescent Proteins PAiRFP1 and PAiRFP2

Biomolecules ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1286 ◽  
Author(s):  
Faez Iqbal Khan ◽  
Fakhrul Hassan ◽  
Razique Anwer ◽  
Feng Juan ◽  
Dakun Lai
Keyword(s):  
Comparative Analysis ◽  
Structural Changes ◽  
Near Infrared ◽  
Fluorescent Proteins ◽  
Irradiation Time ◽  
Fluorescence Emission ◽  
Emission Spectra ◽  
Md Simulations ◽  
Fluorescence Emission Spectra ◽  
Molecular Stability

Two photoactivatable near infrared fluorescent proteins (NIR FPs) named “PAiRFP1” and “PAiRFP2” are formed by directed molecular evolution from Agp2, a bathy bacteriophytochrome of Agrobacterium tumefaciens C58. There are 15 and 24 amino acid substitutions in the structure of PAiRFP1 and PAiRFP2, respectively. A comprehensive molecular exploration of these bacteriophytochrome photoreceptors (BphPs) are required to understand the structure dynamics. In this study, the NIR fluorescence emission spectra for PAiRFP1 were recorded upon repeated excitation and the fluorescence intensity of PAiRFP1 tends to increase as the irradiation time was prolonged. We also predicted that mutations Q168L, V244F, and A480V in Agp2 will enhance the molecular stability and flexibility. During molecular dynamics (MD) simulations, the average root mean square deviations of Agp2, PAiRFP1, and PAiRFP2 were found to be 0.40, 0.49, and 0.48 nm, respectively. The structure of PAiRFP1 and PAiRFP2 were more deviated than Agp2 from its native conformation and the hydrophobic regions that were buried in PAiRFP1 and PAiRFP2 core exposed to solvent molecules. The eigenvalues and the trace of covariance matrix were found to be high for PAiRFP1 (597.90 nm2) and PAiRFP2 (726.74 nm2) when compared with Agp2 (535.79 nm2). It was also found that PAiRFP1 has more sharp Gibbs free energy global minima than Agp2 and PAiRFP2. This comparative analysis will help to gain deeper understanding on the structural changes during the evolution of photoactivatable NIR FPs. Further work can be carried out by combining PCR-based directed mutagenesis and spectroscopic methods to provide strategies for the rational designing of these PAiRFPs.

Hyperspectral imaging as powerful technique for evaluating the stability of Tattoo Wall®

2020 ◽  
Vol 157 ◽  
pp. 104866 ◽  
Author(s):  
G. Agresti ◽  
G. Bonifazi ◽  
G. Capobianco ◽  
L. Lanteri ◽  
C. Pelosi ◽  
...  
Keyword(s):  
Hyperspectral Imaging ◽  
Powerful Technique ◽  
The Stability

scholarly journals Formation of the X-ray line emission spectrum of excimer laser-produced plasmas

1998 ◽  
Vol 16 (1) ◽  
pp. 61-70 ◽  
Author(s):  
A. Magunov ◽  
A. Faenov ◽  
I. Skobelev ◽  
T. Pikuz ◽  
D. Batani ◽  
...  
Keyword(s):  
Experimental Data ◽  
Laser Radiation ◽  
Laser Pulse ◽  
Emission Spectrum ◽  
Pulse Train ◽  
Line Emission ◽  
Emission Spectra ◽  
Theoretical Modelling ◽  
Xecl Laser ◽  
X Ray

Time- and space-integrated emission spectra measurements have been performed in plasma produced by 308 nm wavelength XeCl laser radiation (IL = (4–10)·1012 W/cm2, τ = 10 ns) and by 248 nm wavelength KrF laser pulse train radiation (IL = 5·1015 W/cm2, τ = 7 ps, 16 pulses in train) on CF2 plane target. Theoretical modelling of Lyman series and He-like ion resonance series of fluorine and its fit of experimental data show considerable differences in the absorption of laser radiation in the two plasmas.

scholarly journals Near-Infrared Fluorescent Imaging of Cerebral Thrombi and Blood–Brain Barrier Disruption in a Mouse Model of Cerebral Venous Sinus Thrombosis

2005 ◽  
Vol 25 (2) ◽  
pp. 226-233 ◽  
Author(s):  
Dong-Eog Kim ◽  
Farouc A Jaffer ◽  
Ralph Weissleder ◽  
Ching-Hsuan Tung ◽  
Dawid Schellingerhout
Keyword(s):  
Blood Brain Barrier ◽  
Intravital Microscopy ◽  
Near Infrared ◽  
Thrombus Formation ◽  
Fluorescent Imaging ◽  
Brain Barrier ◽  
Imaging Method ◽  
Microscopy Imaging ◽  
Blood Brain ◽  
Bbb Permeability

An intravital microscopy imaging method was developed to visualize active cerebral thrombus and blood–brain barrier (BBB) disruption using Near Infrared Fluorescent (NIRF) probes. A circular craniotomy was made in CD-1 mice. Thrombi were formed by applying 10%-FeCl3 to the entire exposed superior sagittal sinus (SSS, 5 mm), or to the posterior 2.5 mm of the SSS for 5 mins. Control animals were pretreated with heparin (50 U/kg) before thrombus induction. Three hours after thrombus formation, a FXIIIa-targeted NIRF imaging probe (A15) was intravenously injected, and the SSS was imaged by intravital microscopy. This was followed by injection of indocyanine green (ICG) to assess BBB permeability. The A15 optical probe bound to thrombus, and the fluorescent signal emitted by the bound agent corresponded well with histologically confirmed thrombus. A15 initially remained intravascular, followed by excretion and subsequent decrease in all tissues except for thrombus, where it was retained. The subsequent ICG was also intravascular immediately after injection, but then began to leak into the cerebral parenchyma at 3 to 5 mins. The sites of leakage were adjacent to thrombosed areas. Heparin pretreatment prevented thrombus formation and reduced ICG leakage significantly. This demonstrates the feasibility of simultaneous in vivo monitoring of thrombus and BBB permeability in an animal model of cerebral venous thrombosis.

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