High-Index Optical Materials for High-Throughput Lensfree Imaging, Counting and Sorting of Cells on a Chip

2008 ◽  
Author(s):  
Sungkyu Seo ◽  
Ting-Wei Su ◽  
Anthony Erlinger ◽  
Aydogan Ozcan
Keyword(s):  
High Throughput ◽  
Imaging System ◽  
Signal To Noise Ratio ◽  
Point Of Care ◽  
Cell Types ◽  
Substrate Material ◽  
High Index ◽  
Cell Counting ◽  
Optical Cell ◽  
Numerous Cell

We describe a high-throughput lensfree optical imaging system that can characterize numerous cell types found within a heterogeneous mixture. Here, we specifically discuss the effect of the refractive index of the substrate material on the performance of the proposed lensfree optical cell characterization platform. It is experimentally illustrated that the use of high index materials such as SrTiO3 significantly improves the signal-to-noise ratio of the acquired lensfree images, which is a significant step towards more robust high-throughput cell analysis and characterization. The imaging configuration of the reported high-index material based characterization scheme is massively parallel enabling a cell counting speed of >100,000 cells/sec over a field of view of ∼10 cm2. This novel system can also form the basic building block of a simple and compact point-of-care cell counting technology that can be made as small as a regular cell-phone to rapidly measure the count of e.g., red blood cells or T-lymphocytes from whole blood samples. Such a powerful point-of-care platform may have a significant impact especially for global health related problems in the developing world.

High-Throughput Cell Imaging, Counting and Characterization on a Chip

2008 ◽  
Author(s):  
Ting-Wei Su ◽  
Sungkyu Seo ◽  
Anthony Erlinger ◽  
Aydogan Ozcan
Keyword(s):  
Health Care Providers ◽  
High Throughput ◽  
Point Of Care ◽  
Cell Types ◽  
Field Of View ◽  
Cell Counting ◽  
Care Providers ◽  
Counting Approach ◽  
On Chip

We introduce a lensfree on chip imaging platform that enables high-throughput monitoring, counting, and identification of several different microscopic objects such as different cell types within a heterogeneous solution. This imaging platform can in principle be miniaturized to a hand-held device that can be used by minimally trained health care providers at the point-of-care to measure the cell count of e.g., red blood cells from whole blood samples with a counting speed of >100,000 cells/sec. This novel optical imaging platform can also be merged with microfluidic systems to be able to rapidly monitor and count hundreds of thousand of cells within a field-of-view (FOV) of ∼10 cm2 in vitro. The immediate impact of this lensfree on chip cell counting approach is its improved speed, significantly larger field-of-view and simplified design that permits considerable miniaturization of the entire cell counting device.

scholarly journals A versatile automated high-throughput drug screening platform for zebrafish embryos

2020 ◽  
Author(s):  
Alexandra Lubin ◽  
Jason Otterstrom ◽  
Yvette Hoade ◽  
Ivana Bjedov ◽  
Eleanor Stead ◽  
...  
Keyword(s):  
High Throughput ◽  
Drug Screening ◽  
Imaging System ◽  
Cell Types ◽  
Wide Range ◽  
Stem And Progenitor Cells ◽  
Multiple Cell ◽  
Flexible Imaging ◽  
Automated Screening ◽  
Screening Platform

AbstractZebrafish provide a unique opportunity for drug screening in living animals, with the fast developing, transparent embryos allowing for relatively high throughput, microscopy-based screens. However, the limited availability of rapid, flexible imaging and analysis platforms has limited the use of zebrafish in drug screens. We have developed a easy-to-use, customisable automated screening procedure suitable for high-throughput phenotype-based screens of live zebrafish. We utilised the WiScan®Hermes High Content Imaging System to rapidly acquire brightfield and fluorescent images of embryos, and the WiSoft®Athena Zebrafish Application for analysis, which harnesses an Artificial Intelligence-driven algorithm to automatically detect fish in brightfield images, identify anatomical structures, partition the animal into regions, and exclusively select the desired side-oriented fish. Our initial validation combined structural analysis with fluorescence images to enumerate GFP-tagged haematopoietic stem and progenitor cells in the tails of embryos, which correlated with manual counts. We further validated this system to assess the effects of genetic mutations and x-ray irradiation in high content using a wide range of assays. Further, we performed simultaneous analysis of multiple cell types using dual fluorophores in high throughput. In summary, we demonstrate a broadly applicable and rapidly customisable platform for high content screening in zebrafish.

scholarly journals Actin-binding compounds, previously discovered by FRET-based high-throughput screening, differentially affect skeletal and cardiac muscle

2020 ◽  
Vol 295 (41) ◽  
pp. 14100-14110 ◽  
Author(s):  
Piyali Guhathakurta ◽  
Lien A. Phung ◽  
Ewa Prochniewicz ◽  
Sarah Lichtenberger ◽  
Anna Wilson ◽  
...  
Keyword(s):  
Cardiac Muscle ◽  
High Throughput ◽  
High Throughput Screening ◽  
Cell Types ◽  
Actin Binding ◽  
Time Resolved ◽  
Biochemical Assays ◽  
Numerous Cell ◽  
Cardiac Muscles ◽  
And Function

Actin's interactions with myosin and other actin-binding proteins are essential for cellular viability in numerous cell types, including muscle. In a previous high-throughput time-resolved FRET (TR-FRET) screen, we identified a class of compounds that bind to actin and affect actomyosin structure and function. For clinical utility, it is highly desirable to identify compounds that affect skeletal and cardiac muscle differently. Because actin is more highly conserved than myosin and most other muscle proteins, most such efforts have not targeted actin. Nevertheless, in the current study, we tested the specificity of the previously discovered actin-binding compounds for effects on skeletal and cardiac α-actins as well as on skeletal and cardiac myofibrils. We found that a majority of these compounds affected the transition of monomeric G-actin to filamentous F-actin, and that several of these effects were different for skeletal and cardiac actin isoforms. We also found that several of these compounds affected ATPase activity differently in skeletal and cardiac myofibrils. We conclude that these structural and biochemical assays can be used to identify actin-binding compounds that differentially affect skeletal and cardiac muscles. The results of this study set the stage for screening of large chemical libraries for discovery of novel compounds that act therapeutically and specifically on cardiac or skeletal muscle.

scholarly journals A customizable microscopy system for the automated quantification and characterization of multiple adherent cell types: an alternative to flow cytometry

2018 ◽  
Author(s):  
Vishwaratn Asthana ◽  
Yuqi Tang ◽  
Adam Ferguson ◽  
Pallavi Bugga ◽  
Anantratn Asthana ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Tumor Cells ◽  
Dynamic Range ◽  
Low Cost ◽  
Cell Types ◽  
Cell Counting ◽  
Adherent Cell ◽  
Optical Cell ◽  
Essential Components ◽  
Multiple Cell

Cell quantification assays are essential components of most biological and clinical labs. However, many currently available quantification assays, including flow cytometry and commercial cell counting systems, suffer from unique drawbacks that limit their overall efficacy. In order to address the shortcomings of traditional quantification assays, we have designed a robust, low-cost, automated optical cell cytometer that quantifies individual cells in a multiwell plate using tools readily available in most labs. Plating and subsequent quantification of various dilution series using the automated optical cytometer demonstrates the single-cell sensitivity, near-perfect R2 accuracy, and greater than 5-log dynamic range of our system. Further, the optical cytometer is capable of obtaining absolute counts of multiple cell types in one well as part of a co-culture setup. To demonstrate this ability, we recreated an experiment that assesses the tumoricidal properties of primed macrophages on co-cultured tumor cells as a proof-of-principle test. The results of the experiment reveal that primed macrophages display enhanced cytotoxicity towards tumor cells while simultaneously losing the ability to proliferate, an example of a dynamic interplay between two cell populations that our optical cytometer is successfully able to elucidate.

scholarly journals A customizable microscopy system for the automated quantification and characterization of multiple adherent cell types: an alternative to flow cytometry

2018 ◽  
Author(s):  
Vishwaratn Asthana ◽  
Yuqi Tang ◽  
Adam Ferguson ◽  
Pallavi Bugga ◽  
Anantratn Asthana ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Tumor Cells ◽  
Dynamic Range ◽  
Low Cost ◽  
Cell Types ◽  
Cell Counting ◽  
Adherent Cell ◽  
Optical Cell ◽  
Essential Components ◽  
Multiple Cell

Cell quantification assays are essential components of most biological and clinical labs. However, many currently available quantification assays, including flow cytometry and commercial cell counting systems, suffer from unique drawbacks that limit their overall efficacy. In order to address the shortcomings of traditional quantification assays, we have designed a robust, low-cost, automated optical cell cytometer that quantifies individual cells in a multiwell plate using tools readily available in most labs. Plating and subsequent quantification of various dilution series using the automated optical cytometer demonstrates the single-cell sensitivity, near-perfect R2 accuracy, and greater than 5-log dynamic range of our system. Further, the optical cytometer is capable of obtaining absolute counts of multiple cell types in one well as part of a co-culture setup. To demonstrate this ability, we recreated an experiment that assesses the tumoricidal properties of primed macrophages on co-cultured tumor cells as a proof-of-principle test. The results of the experiment reveal that primed macrophages display enhanced cytotoxicity towards tumor cells while simultaneously losing the ability to proliferate, an example of a dynamic interplay between two cell populations that our optical cytometer is successfully able to elucidate.

scholarly journals A versatile, automated and high-throughput drug screening platform for zebrafish embryos

Biology Open ◽  
2021 ◽  
Author(s):  
Alexandra Lubin ◽  
Jason Otterstrom ◽  
Yvette Hoade ◽  
Ivana Bjedov ◽  
Eleanor Stead ◽  
...  
Keyword(s):  
High Throughput ◽  
Drug Screening ◽  
Imaging System ◽  
Cell Types ◽  
Wide Range ◽  
Stem And Progenitor Cells ◽  
Multiple Cell ◽  
Flexible Imaging ◽  
Automated Screening ◽  
Screening Platform

Zebrafish provide a unique opportunity for drug screening in living animals, with the fast developing, transparent embryos allowing for relatively high-throughput, microscopy-based screens. However, the limited availability of rapid, flexible imaging and analysis platforms has limited the use of zebrafish in drug screens. We have developed an easy-to-use, customisable automated screening procedure suitable for high-throughput phenotype-based screens of live zebrafish. We utilised the WiScan® Hermes High Content Imaging System to rapidly acquire brightfield and fluorescent images of embryos, and the WiSoft® Athena Zebrafish Application for analysis, which harnesses an Artificial Intelligence-driven algorithm to automatically detect fish in brightfield images, identify anatomical structures, partition the animal into regions, and exclusively select the desired side-oriented fish. Our initial validation combined structural analysis with fluorescence images to enumerate GFP-tagged haematopoietic stem and progenitor cells in the tails of embryos, which correlated with manual counts. We further validated this system to assess the effects of genetic mutations and x-ray irradiation in high content using a wide range of assays. Further, we performed simultaneous analysis of multiple cell types using dual fluorophores in high throughput. In summary, we demonstrate a broadly applicable and rapidly customisable platform for high-content screening in zebrafish.

Some Quantification Problems in Parallel EELS Images

1990 ◽  
Vol 48 (2) ◽  
pp. 36-37
Author(s):  
G. Botton ◽  
G. L’Espérance ◽  
M.D. Ball ◽  
C.E. Gallerneault
Keyword(s):  
Electron Microscope ◽  
Imaging System ◽  
Signal To Noise Ratio ◽  
Scanning Transmission Electron Microscope ◽  
Acquisition Time ◽  
Thickness Variation ◽  
Transmission Electron ◽  
Distribution Of Elements ◽  
Scanning Transmission ◽  
Present Distribution

The recently developed parallel electron energy loss spectrometers (PEELS) have led to a significant reduction in spectrum acquisition time making EELS more useful in many applications in material science. Dwell times as short as 50 msec per spectrum with a PEELS coupled to a scanning transmission electron microscope (STEM), can make quantitative EEL images accessible. These images would present distribution of elements with the high spatial resolution inherent to EELS. The aim of this paper is to briefly investigate the effect of acquisition time per pixel on the signal to noise ratio (SNR), the effect of thickness variation and crystallography and finally the energy stability of spectra when acquired in the scanning mode during long periods of time.The configuration of the imaging system is the following: a Gatan PEELS is coupled to a CM30 (TEM/STEM) electron microscope, the control of the spectrometer and microscope is performed through a LINK AN10-85S MCA which is interfaced to a IBM RT 125 (running under AIX) via a DR11W line.

Challenges and Issues in the Design of Micro-Machined Antennas - A Review

2020 ◽  
Vol 13 ◽  
Author(s):  
Ashish Kumar ◽  
Amar Partap Singh Pharwaha
Keyword(s):  
Patch Antenna ◽  
Ground Plane ◽  
Substrate Material ◽  
Review Article ◽  
High Index ◽  
Performance Characteristics ◽  
Machining Process ◽  
Patch Antennas ◽  
Micro Machining ◽  
The Impact

Background: Patch antennas are composed of the substrate material with patch and ground plane on the both sides of the substrate. The dimensions and performance characteristics of the antenna are highly influenced by the choice of the appropriate substrate depending upon the value of their dielectric constant. Generally, low index substrate materials are used to design the patch antenna but there are also some of the applications, which require the implementation of patch antenna design on high index substrate like silicon and gallium arsenide. Objective: The objective of this article is to review the design of antennas developed on high index substrate and the problems associated with the use of these materials as substrate. Also, main challenges and solutions have been discussed to improve the performance characteristics while using the high index substrates. Method: The review article has divided into various sections including the solution of the problems associated with the high index substrates in the form of micro-machining process. Along with this, types of micro machining and their applications have discussed in detail. Results: This review article investigates the various patch antennas designed with micro-machining technology and also discusses the impact of micro-machining process on the performance parameters of the patch antennas designed on high index substrates. Conclusion: By using the micro-machining process, the performance of patch antenna improves drastically but fabrication and tolerances at such minute structures is very tedious task for the antenna designers.

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.

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