scholarly journals Solid State High Throughput Screening Microscopy

2019 ◽  
Author(s):  
M. Ashraf ◽  
S. Mohanan ◽  
B Sim ◽  
A. Tam ◽  
D. Brousseau ◽  
...  
Keyword(s):  
Solid State ◽  
High Throughput ◽  
High Throughput Screening ◽  
Imaging System ◽  
Focal Point ◽  
Collection Efficiency ◽  
Frame Rate ◽  
Single Sample ◽  
Wide Field ◽  
Led Array

We introduce a solid state high throughput screening (ssHTS) imaging modality that uses a novel Newtonian telescope design to image multiple spatially separated samples without moving parts or robotics. Conventional high-throughput imaging modalities either require movement of the sample to the focal plane of the imaging system1–3 or movement of the imaging system itself4,5, or use a wide-field approach to capture several samples in one frame. Schemes which move the sample or the imaging system can be mechanically complex and are inherently slow, while wide-field imaging systems have poor light collection efficiency and resolution compared to systems that image a single sample at a given time point. Our proposed ssHTS system uses a large parabolic reflector and an imaging lenses positioned at their focal distances above each sample. A fast LED array sequentially illuminate samples to generate images that are captured with a single camera placed at the focal point of the reflector. This optical configuration allows each sample to completely fill a sensors field of view. Since each LED illuminates a single sample and LED switch times are very fast, images from spatially separated samples can be captured at rates limited only by the camera’s frame rate. The system is demonstrated by imaging cardiac monolayer and Caenorhabditis elegans (C. elegans) preparations.

scholarly journals Development of a Fluorescence-Based, Ultra High-Throughput Screening Platform for Nanoliter-Scale Cytochrome P450 Microarrays

2009 ◽  
Vol 14 (6) ◽  
pp. 668-678 ◽  
Author(s):  
Sumitra M. Sukumaran ◽  
Benjamin Potsaid ◽  
Moo-Yeal Lee ◽  
Douglas S. Clark ◽  
Jonathan S. Dordick
Keyword(s):  
Enzyme Activity ◽  
Cytochrome P450 ◽  
High Throughput ◽  
High Throughput Screening ◽  
Imaging System ◽  
Early Stage ◽  
Wide Field ◽  
Cytochrome P450 Enzyme ◽  
Assay Sensitivity ◽  
Screening Platform

Cytochrome P450 enzyme (CYP450s) assays are critical enzymes in early-stage lead discovery and optimization in drug development. Currently available fluorescence-based reaction assays provide a rapid and reliable method for monitoring CYP450 enzyme activity but are confined to medium-throughput well-plate systems. The authors present a high-throughput, integrated screening platform for CYP450 assays combining enzyme encapsulation techniques, microarraying methods, and wide-field imaging. Alginate-containing microarrays consisting of up to 1134 CYP450 reaction elements were fabricated on functionalized glass slides (reaction volumes 20 to 80 nL, total enzyme content in pg) and imaged to yield endpoint activity, stability, and kinetic data. A charge-coupled device imager acquired quantitative, high-resolution images of a 20 × 20 mm area/snapshot using custom-built wide-field optics with telecentric lenses and easily interchangeable filter sets. The imaging system offered a broad dynamic intensity range (linear over 3 orders of magnitude) and sensitivity down to fluorochrome quantities of <5 fmols, with read accuracy similar to a laser scanner or a fluorescence plate reader but with higher throughput. Rapid image acquisition enabled analysis of CYP450 kinetics. Fluorogenic assays with CYP3A4, CYP2C9, and CYP2D6 on the alginate microarrays exhibited Z′ factors ranging from 0.75 to 0.85, sensitive detection of inhibitory compounds, and reactivity comparable to that in solution, thereby demonstrating the reliability and accuracy of the microarray platform. This system enables for the first time a significant miniaturization of CYP enzyme assays with significant conservation of assay reagents, greatly increased throughput, and no apparent loss of enzyme activity or assay sensitivity. ( Journal of Biomolecular Screening 2009:668-678)

scholarly journals A high-throughput screening assay based on automated microscopy for monitoring antibiotic susceptibility of Mycobacterium tuberculosis phenotypes

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sadaf Kalsum ◽  
Blanka Andersson ◽  
Jyotirmoy Das ◽  
Thomas Schön ◽  
Maria Lerm
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
High Throughput Screening ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging System ◽  
Aggregate Size ◽  
Live Cell ◽  
Screening Assay ◽  
High Throughput Screening Assay

Abstract Background Efficient high-throughput drug screening assays are necessary to enable the discovery of new anti-mycobacterial drugs. The purpose of our work was to develop and validate an assay based on live-cell imaging which can monitor the growth of two distinct phenotypes of Mycobacterium tuberculosis and to test their susceptibility to commonly used TB drugs. Results Both planktonic and cording phenotypes were successfully monitored as fluorescent objects using the live-cell imaging system IncuCyte S3, allowing collection of data describing distinct characteristics of aggregate size and growth. The quantification of changes in total area of aggregates was used to define IC50 and MIC values of selected TB drugs which revealed that the cording phenotype grew more rapidly and displayed a higher susceptibility to rifampicin. In checkerboard approach, testing pair-wise combinations of sub-inhibitory concentrations of drugs, rifampicin, linezolid and pretomanid demonstrated superior growth inhibition of cording phenotype. Conclusions Our results emphasize the efficiency of using automated live-cell imaging and its potential in high-throughput whole-cell screening to evaluate existing and search for novel antimycobacterial drugs.

scholarly journals Bionic Birdlike Imaging Using a Multi-Hyperuniform LED Array

Sensors ◽  
2021 ◽  
Vol 21 (12) ◽  
pp. 4084
Author(s):  
Xin-Yu Zhao ◽  
Li-Jing Li ◽  
Lei Cao ◽  
Ming-Jie Sun
Keyword(s):  
Imaging System ◽  
Color Image ◽  
Digital Camera ◽  
Frame Rate ◽  
Digital Cameras ◽  
Structured Illumination ◽  
Pixel Resolution ◽  
Led Array ◽  
Frequency Aliasing ◽  
Imaging Sensors

Digital cameras obtain color information of the scene using a chromatic filter, usually a Bayer filter, overlaid on a pixelated detector. However, the periodic arrangement of both the filter array and the detector array introduces frequency aliasing in sampling and color misregistration during demosaicking process which causes degradation of image quality. Inspired by the biological structure of the avian retinas, we developed a chromatic LED array which has a geometric arrangement of multi-hyperuniformity, which exhibits an irregularity on small-length scales but a quasi-uniformity on large scales, to suppress frequency aliasing and color misregistration in full color image retrieval. Experiments were performed with a single-pixel imaging system using the multi-hyperuniform chromatic LED array to provide structured illumination, and 208 fps frame rate was achieved at 32 × 32 pixel resolution. By comparing the experimental results with the images captured with a conventional digital camera, it has been demonstrated that the proposed imaging system forms images with less chromatic moiré patterns and color misregistration artifacts. The concept proposed verified here could provide insights for the design and the manufacturing of future bionic imaging sensors.

scholarly journals Beeporter: a high-throughput tool for non-invasive analysis of honey bee virus infection

2021 ◽  
Author(s):  
Jay D. Evans ◽  
Olubukola Banmeke ◽  
Evan C. Palmer-Young ◽  
Yanping Chen ◽  
Eugene V. Ryabov
Keyword(s):  
Virus Infection ◽  
Honey Bee ◽  
High Throughput ◽  
Honey Bees ◽  
High Throughput Screening ◽  
Fluorescent Protein ◽  
Imaging System ◽  
Uv Light ◽  
Light Sources ◽  
Deformed Wing Virus

ABSTRACTHoney bees face numerous pests and pathogens but arguably none are as devastating as Deformed wing virus (DWV). Development of antiviral therapeutics and virus-resistant honey bee lines to control DWV in honey bees is slowed by the lack of a cost-effective high-throughput screening of DWV infection. Currently, analysis of virus infection and screening for antiviral treatments in bees and their colonies is tedious, requiring a well-equipped molecular biology laboratory and the use of hazardous chemicals. Here we utilize a cDNA clone of DWV tagged with green fluorescent protein (GFP) to develop the Beeporter assay, a method for detection and quantification of DWV infection in live honey bees. The assay involves infection of honey bee pupae by injecting a standardized DWV-GFP inoculum, followed by incubation for up to 44 hours. GFP fluorescence is recorded at intervals via commonly available long-wave UV light sources and a smartphone camera or a standard ultraviolet transilluminator gel imaging system. Nonlethal DWV monitoring allows high-throughput screening of antiviral candidates and a direct breeding tool for identifying honey bee parents with increased antivirus resistance. For even more rapid drug screening, we also describe a method for screening bees using 96-well trays and a spectrophotometer.

scholarly journals Electro-optic imaging enables efficient wide-field fluorescence lifetime microscopy

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Adam J. Bowman ◽  
Brannon B. Klopfer ◽  
Thomas Juffmann ◽  
Mark A. Kasevich
Keyword(s):  
Temporal Resolution ◽  
Single Molecule ◽  
Fluorescence Lifetime ◽  
Collection Efficiency ◽  
Frame Rate ◽  
Optical Systems ◽  
Wide Field ◽  
Pockels Cell ◽  
Low Light ◽  
Optical Efficiency

Abstract Nanosecond temporal resolution enables new methods for wide-field imaging like time-of-flight, gated detection, and fluorescence lifetime. The optical efficiency of existing approaches, however, presents challenges for low-light applications common to fluorescence microscopy and single-molecule imaging. We demonstrate the use of Pockels cells for wide-field image gating with nanosecond temporal resolution and high photon collection efficiency. Two temporal frames are obtained by combining a Pockels cell with a pair of polarizing beam-splitters. We show multi-label fluorescence lifetime imaging microscopy (FLIM), single-molecule lifetime spectroscopy, and fast single-frame FLIM at the camera frame rate with 103–105 times higher throughput than single photon counting. Finally, we demonstrate a space-to-time image multiplexer using a re-imaging optical cavity with a tilted mirror to extend the Pockels cell technique to multiple temporal frames. These methods enable nanosecond imaging with standard optical systems and sensors, opening a new temporal dimension for wide-field low-light microscopy.

High-throughput screening of solid-state catalyst libraries

Nature ◽  
10.1038/28575 ◽  
1998 ◽  
Vol 394 (6691) ◽  
pp. 350-353 ◽  
Author(s):  
Selim M. Senkan
Keyword(s):  
Solid State ◽  
High Throughput ◽  
High Throughput Screening

scholarly journals High-Throughput Screening of Encapsulated Islets Using Wide-Field Lens-Free On-Chip Imaging

ACS Photonics ◽  
2018 ◽  
Vol 5 (6) ◽  
pp. 2081-2086 ◽  
Author(s):  
Yibo Zhang ◽  
Michael Alexander ◽  
Sam Yang ◽  
Yinxu Bian ◽  
Elliot Botvinick ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Wide Field ◽  
On Chip

scholarly journals Dendritic polymer imaging systems for the evaluation of conjugate uptake and cleavage

Nanoscale ◽  
2015 ◽  
Vol 7 (9) ◽  
pp. 3838-3844 ◽  
Author(s):  
Harald R. Krüger ◽  
Gregor Nagel ◽  
Stefanie Wedepohl ◽  
Marcelo Calderón
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Imaging System ◽  
Imaging Systems ◽  
Microplate Assay ◽  
Dendritic Polymer ◽  
Intracellular Release ◽  
A Cell

A FRET-based imaging system was developed to evaluate polymer uptake and intracellular release of cargo in a cell based microplate assay that is suitable for high throughput screening.

scholarly journals Automated High Throughput Screening for Serine Kinase Inhibitors Using a LEADSeeker™ Scintillation Proximity Assay in the 1536-Well Format

2002 ◽  
Vol 7 (1) ◽  
pp. 11-19 ◽  
Author(s):  
Gabriele Sorg ◽  
Hans-Dieter Schubert ◽  
Frank H. Büttner ◽  
Ralf Heilker
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Kinase Inhibitors ◽  
Imaging System ◽  
Serine Kinase ◽  
Response Curves ◽  
Scintillation Proximity Assay ◽  
Inhibitory Compounds ◽  
Well Assay ◽  
Biotinylated Peptide

High-throughput screening in the 1536-well format has been largely restricted to solution-based and cell-based screens. In this article, we show the feasibility of a completely automated, robust scintillation proximity assay in the 1536-well format that is suitable to identify inhibitors for a serine/threonine kinase from a compound library. The introduction of [33P]phosphate into a biotinylated peptide substrate mirrors the activity of the kinase. The peptide is immobilized on streptavidin-coated LEADseeker imaging beads and [33P]phosphate incorporation is detected with the LEADseeker imaging system of Amersham Pharmacia Biotech. To improve the liquid handling procedures for imaging bead suspensions in the low microliter range, we developed a novel trough with an integrated stirring function. A comparison of the 1536-well assay to a 384-well assay revealed a comparable assay quality with Z’ factors of about 0.7 for the 384-well format and 0.6 for the 1536-well format. In an automated screen of a random compound collection, 94.4% of the inhibitory compounds could be identified with both assay formats. Dose-response curves were performed for a selection of identified kinase inhibitors and revealed similar IC50 values for both assay formats.

scholarly journals A fluorescence-based high-throughput screening method for cytokinin translocation mutants

Plant Methods ◽  
2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Mengyuan Zhang ◽  
Bingli Ding ◽  
Jiangzhe Zhao ◽  
Penghong Zhang ◽  
Yujia Li ◽  
...  
Keyword(s):  
High Throughput ◽  
Stress Responses ◽  
High Throughput Screening ◽  
Large Scale ◽  
Fluorescent Protein ◽  
Imaging System ◽  
Screening Method ◽  
Cytokinin Signaling ◽  
Phenotypic Analysis ◽  
Long Distance

Abstract Background Cytokinins are one kind of phytohormones essential for plant growth, development and stress responses. In the past half century, significant progresses have been made in the studies of cytokinin signal transduction and metobolic pathways, but the mechanism of cytokinin translocation is poorly understood. Arabidopsis (Arabidopsis thaliana) response regulator 5 (ARR5) is a type-A response factor in cytokinin signaling which is induced by cytokinins and has been used as a reporter gene for the endogenous cytokinins in Arabidopsis. Here, we report a fluorescence-based high-throughput method to screen cytokinin translocation mutants using an ethyl methyl sulfone (EMS) mutagenesis library generated with ARR5::eGFP transgenic plants. Results The seedlings with enhanced green fluorescent protein (GFP) signal in roots were screened in a luminescence imaging system (LIS) in large scale to obtain mutants with over-accumulated cytokinins in roots. The selected mutants were confirmed under a fluorescence microscopy and then performed phenotypic analysis. In this way, we obtained twelve mutants with elevated GFP signal in the roots and further found three of them displayed reduced GFP signal in the aerial tissues. Two of the mutants were characterized and proved to be the atabcg14 allelic mutants which are defective in the long-distance translocation of root-synthesized cytokinins. Conclusions We provide a strategy for screening mutants defective in cytokinin translocation, distribution or signaling. The strategy can be adapted to establish a system for screening mutants defective in other hormone transporters or signaling components using a fluorescence reporter.

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