Atto 465 Derivative Is a Nuclear Stain with Unique Excitation and Emission Spectra Useful for Multiplex Immunofluorescence Histochemistry

2022 ◽  
pp. 002215542110649
Author(s):  
Joshua T. Dodge ◽  
Andrew D. Doyle ◽  
Ana C. Costa-da-Silva ◽  
Christopher T. Hogden ◽  
Eva Mezey ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Fluorescent Probe ◽  
Tissue Section ◽  
Fluorescent Dyes ◽  
Emission Spectra ◽  
Visible Spectrum ◽  
Multiple Target ◽  
Effective Technique ◽  
Fluorescence Excitation

Multiplex immunofluorescence (mIF) is an effective technique for the maximal visualization of multiple target proteins in situ. This powerful tool is mainly limited by the spectral overlap of the currently available synthetic fluorescent dyes. The fluorescence excitation wavelengths ranging between 405 and 488 nm are rarely used in mIF imaging and serve as a logical additional slot for a fluorescent probe. In the present study, we demonstrate that the addition of 2,3,4,5,6-pentafluoroaniline to Atto 465 NHS ester, creating Atto 465-pentafluoroaniline (Atto 465-p), generates a bright nuclear stain in the violet-blue region of the visible spectrum. This allows the 405 nm excitation and emission, classically used for nuclear counterstains, to be used for the detection of another target protein. This increases the flexibility of the mIF panel and, with appropriate staining and microscopy, enables the quantitative analysis of at least six targets in one tissue section. (J Histochem Cytochem XX: XXX–XXX, XXXX)

scholarly journals Atto 465 derivative is a nuclear stain with unique excitation and emission spectra for multiplex immunofluorescence histochemistry

2021 ◽  
Author(s):  
Joshua T Dodge ◽  
Ana Caroline Costa-da-Silva ◽  
Christopher T Hogden ◽  
Eva Mezey ◽  
Jacqueline W Mays
Keyword(s):  
Quantitative Analysis ◽  
Fluorescent Dyes ◽  
Emission Spectra ◽  
Visible Spectrum ◽  
Multiple Target ◽  
Tissue Architecture ◽  
Effective Technique ◽  
Fluorescence Excitation ◽  
Biologic Response

The architecture of a biologic response is inextricably linked with the tissue architecture of the target site. Multiplex immunofluorescence (mIF) is an effective technique for the maximal visualization of multiple target proteins in situ. This powerful tool is limited by the spectral overlap separation of the currently available synthetic fluorescent dyes. The fluorescence excitation wavelengths ranging between 405nm and 488nm are rarely used in mIF imaging and serve as a logical additional slot for a fluorescent probe. In the present study, we demonstrate that Atto 465-pentafluoroaniline (Atto 465-p), a derivative of the fluorescent dye Atto 465, can serve as a nuclear stain in the violet-blue region of the visible spectrum. This opens the 405nm channel, traditionally used for nuclear stain, for detection of another experimental target. This increases the flexibility of the mIF panel and, with appropriate staining and microscopy, enables the quantitative analysis of at least six targets in one tissue section.

Visible and Ultraviolet Absorption and Fluorescence Excitation and Emission Characteristics of Florida Orange Juice and Orange Pulpwash: Detection of Adulteration

1980 ◽  
Vol 63 (6) ◽  
pp. 1317-1331 ◽  
Author(s):  
Donald R Petrus ◽  
John A Attaway
Keyword(s):  
Orange Juice ◽  
Emission Spectra ◽  
Visible Spectrum ◽  
Linear Increase ◽  
Ultraviolet Absorption ◽  
Absorption Data ◽  
Fluorescence Excitation ◽  
Maximum Emission ◽  
Absorbance Ratio ◽  
Florida Citrus

Abstract Visible and ultraviolet absorption and room temperature fluorescence excitation and emission spectra, obtained from alcoholic solutions of Florida orange juice and pulpwash, were investigated to characterize citrus products, and thereby detect adulteration and approximate orange juice content. Absorption shoulders and maxima were observed at 465, 443, and 425 nm of the visible spectrum and at 325, 280, and 245 nm of the ultraviolet spectrum. The spectral curves for orange juice and pulpwash were similar. However, orange pulpwash had weaker visible and stronger ultraviolet absorption than did orange juice. It was determined (416 frozen concentrated orange juice and 206 orange pulpwash samples obtained from Florida commercial processors) that the average sum and standard deviation of absorbance at 443, 325, and 280 nm was 2.410 ± 0.164 for orange juice and 3.781 ± 0.473 for orange pulpwash (based on reconstituted 12.8° Brix products). Average absorbance 443/325 nm ratio was 0.144 ± 0.026 for orange juice and 0.048 ± 0.020 for orange pulpwash. Simple dilutions of orange juice with water and/or sugar solutions resulted in a linear decrease in absorbance sum while the absorbance ratio remained essentially the same. Orange juice and pulpwash systems resulted in a linear increase in absorbance sum and a decrease in absorbance ratio (natural log function). The investigation enabled the derivation of equations, using absorption data, to approximate orange juice and/or pulpwash contents and/or dilution of a sample. The constants used in the equations were based on results obtained from commercially produced Florida citrus products and may vary with other citrus-producing regions. Fluorescence excitation and emission spectra complemented the absorption spectra. Fluorescence excitation for orange juice was maximum at 283 nm with a shoulder at 291 and inflections at 270 and 302 nm with maximum emission at 333 nm. Excitation was also maximum at 291 nm with a shoulder at 283 and inflections at 270 and 302 with maximum emission at 343 nm. Pulpwash excitation was maximum at 270 and 302 nm with a minimum at 283 nm and emission at 353 nm. Peaks or pronounced shoulders at 270 and 302 nm have been indicative of the presence of orange pulpwash.

A fluorescent probe based on N-doped carbon dots for highly sensitive detection of Hg2+ in aqueous solutions

2016 ◽  
Vol 8 (10) ◽  
pp. 2297-2304 ◽  
Author(s):  
Zhi-hao Gao ◽  
Zheng-zhong Lin ◽  
Xiao-mei Chen ◽  
Hui-ping Zhong ◽  
Zhi-yong Huang
Keyword(s):  
Absorption Spectrum ◽  
Aqueous Solutions ◽  
Fluorescent Probe ◽  
Carbon Dots ◽  
Emission Spectra ◽  
Sensitive Detection ◽  
Fluorescence Excitation ◽  
Highly Sensitive ◽  
Highly Sensitive Detection ◽  
Doped Carbon Dots

The UV-Vis absorption spectrum (left) and fluorescence excitation and emission spectra (right) of NCDs.

Immunoassay techniques with fluorescent phycobiliprotein conjugates.

1983 ◽  
Vol 29 (9) ◽  
pp. 1582-1586 ◽  
Author(s):  
M N Kronick ◽  
P D Grossman
Keyword(s):  
Spectral Properties ◽  
Fluorescent Dyes ◽  
Solid Phase ◽  
Visible Spectrum ◽  
The Other ◽  
Cross Linking ◽  
The Novel ◽  
Sandwich Assay ◽  
Fluorescence Excitation ◽  
High Quantum

Abstract We describe immunoassay techniques that take advantage of the novel properties of phycobiliprotein fluorescent dyes. These dyes, which can be isolated from a wide variety of algae, exhibit extremely high absorptivities, high quantum efficiencies, and excitation and emission bands across the visible spectrum. These stable, hydrophilic proteins can easily be linked to antibodies by conventional protein cross-linking reagents. One assay technique presented here demonstrates the use of phycobiliproteins in a solid-phase "sandwich" assay, which achieves picomolar sensitivity. In the other technique the unique spectral properties of phycobiliproteins are applied to fluorescence excitation transfer immunoassay.

Turn-on fluorescent probe for dopamine detection in solutions and live cells based on in situ formation of aminosilane-functionalized carbon dots

2021 ◽  
Vol 1157 ◽  
pp. 338394
Author(s):  
Xiao-Yue Tang ◽  
Yi-Ming Liu ◽  
Xiao-Lin Bai ◽  
Hao Yuan ◽  
Yi-Kao Hu ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Carbon Dots ◽  
Live Cells ◽  
Turn On ◽  
Dopamine Detection ◽  
In Situ Formation

Rabbit aortic medial thickness under relaxed and specified simulated in vivo conditions

1983 ◽  
Vol 245 (4) ◽  
pp. H623-H627
Author(s):  
G. Schneiderman ◽  
W. F. Pritchard ◽  
C. A. Ramirez ◽  
C. K. Colton ◽  
K. A. Smith ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Mean Arterial Pressure ◽  
Aortic Wall ◽  
Macromolecular Transport ◽  
Liquid Silicone ◽  
Normal Mean ◽  
Longitudinal Extension ◽  
Media Layer

A method is presented for measuring the thickness of the intima-media layer of the normal rabbit descending thoracic aortic wall under both relaxed (excised) and specified simulated in vivo conditions. The in vivo conditions were simulated by maintaining the aorta in situ at its normal longitudinal extension while perfusing its lumen at the normal mean arterial pressure with a mixture of liquid silicone polymer and a catalyst, thus providing physiological radial distension. After the rubber cured, both relaxed and extended-distended tissue segments were obtained from adjacent sites on the same aorta. These tissue segments were fixed and further processed for measurement of their medial thicknesses by light microscopy. This data was used to estimate the ratio of the medial thickness of the relaxed, excised aorta to that under in vivo conditions, 1.72 +/- 0.15. This information is required for quantitative analysis of data obtained from previous studies of in vivo macromolecular transport across the rabbit thoracic aortic wall.

C-fos and c-jun Proto-Oncogene Expression Is Decreased in Psoriasis: an In Situ Quantitative Analysis

1991 ◽  
Vol 97 (4) ◽  
pp. 672-678 ◽  
Author(s):  
Nicole. Basset-Séguin ◽  
Chantal. Escot ◽  
Jean Pierre. Molès ◽  
Jean Marie. Blanchard ◽  
Cécile. Kerai ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Oncogene Expression

scholarly journals Analyzing Satellite Ocean Color Match-Up Protocols Using the Satellite Validation Navy Tool (SAVANT) at MOBY and Two AERONET-OC Sites

2021 ◽  
Vol 13 (14) ◽  
pp. 2673
Author(s):  
Adam Lawson ◽  
Jennifer Bowers ◽  
Sherwin Ladner ◽  
Richard Crout ◽  
Christopher Wood ◽  
...  
Keyword(s):  
Infrared Imaging ◽  
Ocean Color ◽  
Visible Spectrum ◽  
Ground Truth ◽  
Naval Research ◽  
Satellite Validation ◽  
The Difference ◽  
Stability And Accuracy ◽  
Satellite Ocean Color

The satellite validation navy tool (SAVANT) was developed by the Naval Research Laboratory to help facilitate the assessment of the stability and accuracy of ocean color satellites, using numerous ground truth (in situ) platforms around the globe and support methods for match-up protocols. The effects of varying spatial constraints with permissive and strict protocols on match-up uncertainty are evaluated, in an attempt to establish an optimal satellite ocean color calibration and validation (cal/val) match-up protocol. This allows users to evaluate the accuracy of ocean color sensors compared to specific ground truth sites that provide continuous data. Various match-up constraints may be adjusted, allowing for varied evaluations of their effects on match-up data. The results include the following: (a) the difference between aerosol robotic network ocean color (AERONET-OC) and marine optical Buoy (MOBY) evaluations; (b) the differences across the visible spectrum for various water types; (c) spatial differences and the size of satellite area chosen for comparison; and (d) temporal differences in optically complex water. The match-up uncertainty analysis was performed using Suomi National Polar-orbiting Partnership (SNPP) Visible Infrared Imaging Radiometer Suite (VIIRS) SNPP data at the AERONET-OC sites and the MOBY site. It was found that the more permissive constraint sets allow for a higher number of match-ups and a more comprehensive representation of the conditions, while the restrictive constraints provide better statistical match-ups between in situ and satellite sensors.

Wrinkle and Near Resonance Effect on the Vibrational and Electronic properties in Compressed Monolayer MoSe2

2021 ◽  
Author(s):  
Yan Liu ◽  
Qiang Zhou ◽  
Yalan Yan ◽  
Liang Li ◽  
Jian Zhu ◽  
...  
Keyword(s):  
High Pressure ◽  
Optical Properties ◽  
Transition Metal ◽  
Electronic Properties ◽  
Resonance Effect ◽  
Transition Metal Dichalcogenides ◽  
Effective Technique ◽  
Near Resonance ◽  
Metal Dichalcogenides

Pressure has been considered as an effective technique to modulate the structural, electronic, and optical properties of transition metal dichalcogenides (TMDs) materials. Here, by performing in situ high pressure Raman,...

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