scholarly journals Sequentially addressable dielectrophoretic array for high-throughput sorting of large-volume biological compartments

2020 ◽  
Vol 6 (22) ◽  
pp. eaba6712 ◽  
Author(s):  
A. Isozaki ◽  
Y. Nakagawa ◽  
M. H. Loo ◽  
Y. Shibata ◽  
N. Tanaka ◽  
...  
Keyword(s):  
Single Cell ◽  
High Speed ◽  
Single Cell Analysis ◽  
Droplet Microfluidics ◽  
Cell Analysis ◽  
Green Biotechnology ◽  
Speed Flow ◽  
Droplet Sorting ◽  
On Chip

Droplet microfluidics has become a powerful tool in precision medicine, green biotechnology, and cell therapy for single-cell analysis and selection by virtue of its ability to effectively confine cells. However, there remains a fundamental trade-off between droplet volume and sorting throughput, limiting the advantages of droplet microfluidics to small droplets (<10 pl) that are incompatible with long-term maintenance and growth of most cells. We present a sequentially addressable dielectrophoretic array (SADA) sorter to overcome this problem. The SADA sorter uses an on-chip array of electrodes activated and deactivated in a sequence synchronized to the speed and position of a passing target droplet to deliver an accumulated dielectrophoretic force and gently pull it in the direction of sorting in a high-speed flow. We use it to demonstrate large-droplet sorting with ~20-fold higher throughputs than conventional techniques and apply it to long-term single-cell analysis of Saccharomyces cerevisiae based on their growth rate.

scholarly journals Optimized double emulsion flow cytometry with high-throughput single droplet isolation

2019 ◽  
Author(s):  
Kara K. Brower ◽  
Catherine Carswell-Crumpton ◽  
Sandy Klemm ◽  
Bianca Cruz ◽  
Gaeun Kim ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Single Cell Analysis ◽  
Double Emulsion ◽  
Droplet Microfluidics ◽  
Cell Analysis ◽  
Single Droplet ◽  
Emulsion Droplets ◽  
Droplet Sorting

Droplet microfluidics has made large impacts in diverse areas such as enzyme evolution, chemical product screening, polymer engineering, and single-cell analysis. However, while droplet reactions have become increasingly sophisticated, phenotyping droplets by a fluorescent signal and sorting them to isolate variants-of-interest remains a field-wide bottleneck. Here, we present an optimized double emulsion workflow, sdDE-FACS, that enables high-throughput phenotyping, selection, and sorting of droplets using standard flow cytometers. Using a 130 μm nozzle, we demonstrate robust post-sort recovery of intact droplets, with little to no shear-induced droplet breakage, at high sort frequency (12-14 kHz) across two industry-standard FACS instruments. We report the first quantitative plate statistics for double emulsion droplet isolation and demonstrate single droplet recovery with >70% efficiency. In addition, we establish complete downstream recovery of nucleic acids from single, sorted double emulsion droplets, an advance in droplet sorting comparable with the capabilities of single-cell FACS. This work resolves several hurdles in the field of high-throughput droplet analysis and paves the way for a variety of new droplet assays, including rare variant isolation and multiparameter single-cell analysis, marrying the full power of flow cytometry with droplet microfluidics.

scholarly journals More than efficacy revealed by single-cell analysis of antiviral therapeutics

2019 ◽  
Author(s):  
Wu Liu ◽  
Mehmet U. Caglar ◽  
Zhangming Mao ◽  
Andrew Woodman ◽  
Jamie J. Arnold ◽  
...  
Keyword(s):  
Single Cell ◽  
Drug Targets ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Principal Component ◽  
Viral Population ◽  
Cell Analysis ◽  
Toxic Dose ◽  
Time Required

SUMMARYDevelopment of antiviral therapeutics emphasizes minimization of the effective dose and maximization of the toxic dose, first in cell culture and later in animal models. Long-term success of an antiviral therapeutic is determined not only by its efficacy but also by the duration of time required for drug-resistance to evolve. We have developed a microfluidic device comprised of ~6000 wells, with each well containing a microstructure to capture single cells. We have used this device to characterize enterovirus inhibitors with distinct mechanisms of action. In contrast to population methods, single-cell analysis reveals that each class of inhibitor interferes with the viral infection cycle in a manner that can be distinguished by principal component analysis. Single-cell analysis of antiviral candidates reveals not only efficacy but also properties of the members of the viral population most sensitive to the drug, the stage of the lifecycle most affected by the drug, and perhaps even if the drug targets an interaction of the virus with its host.

scholarly journals Massively parallel quantification of phenotypic heterogeneity in single cell drug responses

2020 ◽  
Author(s):  
Benjamin B. Yellen ◽  
Jon S. Zawistowski ◽  
Eric A. Czech ◽  
Caleb I. Sanford ◽  
Elliott D. SoRelle ◽  
...  
Keyword(s):  
Machine Learning ◽  
Acute Myeloid Leukemia ◽  
Single Cell ◽  
Myeloid Leukemia ◽  
Single Cell Analysis ◽  
Phenotypic Heterogeneity ◽  
Massively Parallel ◽  
Cell Analysis ◽  
On Chip ◽  
Acute Myeloid

AbstractSingle cell analysis tools have made significant advances in characterizing genomic heterogeneity, however tools for measuring phenotypic heterogeneity have lagged due to the increased difficulty of handling live biology. Here, we report a single cell phenotyping tool capable of measuring image-based clonal properties at scales approaching 100,000 clones per experiment. These advances are achieved by exploiting a novel flow regime in ladder microfluidic networks that, under appropriate conditions, yield a mathematically perfect cell trap. Machine learning and computer vision tools are used to control the imaging hardware and analyze the cellular phenotypic parameters within these images. Using this platform, we quantified the responses of tens of thousands of single cell-derived acute myeloid leukemia (AML) clones to targeted therapy, identifying rare resistance and morphological phenotypes at frequencies down to 0.05%. This approach can be extended to higher-level cellular architectures such as cell pairs and organoids and on-chip live-cell fluorescence assays.

(Invited) On-Chip Characterization of Microcapsules Using a Capacitive Sensor for Microencapsulation and Single-Cell Analysis Applications

2021 ◽  
Vol MA2021-01 (60) ◽  
pp. 1603-1603
Author(s):  
Sajjad Janfaza ◽  
Seyedehhamideh Razavi ◽  
Arash Dalili ◽  
Mina Hoorfar
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Capacitive Sensor ◽  
Cell Analysis ◽  
On Chip

MAGNETOPHORETIC CIRCUIT BIOCOMPATIBILITY

2020 ◽  
Vol 20 (07) ◽  
pp. 2050050
Author(s):  
ROOZBEH ABEDINI-NASSAB
Keyword(s):  
Shear Stress ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Cell Behavior ◽  
Cell Analysis ◽  
Applied Shear Stress ◽  
Cell Handling ◽  
Magnetic Labeling ◽  
Long Term Studies

Recently, we introduced magnetophoretic circuits, composed of overlaid magnetic and metallic layers, as a novel single-cell analysis (SCA) tool. We showed the ability of these circuits in organizing large single-particle and particle-pair arrays. Assembling the cells in microarrays is performed with the ultimate goal of running temporal phenotypic analyses. However, for long-term studies, a suitable microenvironment for the cells to normally grow and differentiate is needed. Towards this goal, in this study, we run required biocompatibility tests, based on which we make the magnetophoretic-based microchip a suitable home for the cells to grow. The results confirm the ability of these chips in cell handling and show no unwanted cell behavior alteration due to the applied shear stress on them, the magnetic labeling, or the microenvironment. After this achievement, this tool would be ready for running important single-cell studies in oncology, virology, and medicine.

High-Throughput, Long-Term Imaging of Salmonella Infecting Macrophages in a Micro-Environmental System

2006 ◽  
Author(s):  
Shih-Hui Chao ◽  
Tim J. Strovas ◽  
Ting-She M. Wang ◽  
Kendan A. Jones-Isaac ◽  
Susan L. Fink ◽  
...  
Keyword(s):  
Single Cell ◽  
Real Time ◽  
Laser Scanning ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Cellular Functions ◽  
Laser Scanning Confocal Microscope ◽  
History Of ◽  
Multiple Variables

Real-time single cell analysis is necessary to understand dynamic cellular functions in time and space. Such analyses require the simultaneous measurement of multiple variables in real-time, due to heterogeneity in cellular populations. We report the application of using a micro-environmental chamber on an automatic laser scanning confocal microscope to observe murine macrophage cells in incubation conditions for more than 18 hours. The motorized stage of the microscope was programmed to scan through pre-defined monitoring locations to increase the observation throughput. The acquired images were post-processed to extract the information of each cell. In contrast to current single-cell technologies, such as fluorescence-activated cell sorter (FACS) based systems, the reported architecture records the history of the physiological responses of individual cells.

scholarly journals Development of Droplet Microfluidics Enabling High-Throughput Single-Cell Analysis

Molecules ◽  
2016 ◽  
Vol 21 (7) ◽  
pp. 881 ◽  
Author(s):  
Na Wen ◽  
Zhan Zhao ◽  
Beiyuan Fan ◽  
Deyong Chen ◽  
Dong Men ◽  
...  
Keyword(s):  
Single Cell ◽  
High Throughput ◽  
Single Cell Analysis ◽  
Droplet Microfluidics ◽  
Cell Analysis
Sign in / Sign up

Export Citation Format

Share Document