scholarly journals Single Cell Analysis of Neutrophils NETs by Microscopic LSPR Imaging System

Micromachines ◽  
2019 ◽  
Vol 11 (1) ◽  
pp. 52 ◽  
Author(s):  
Riyaz Ahmad Mohamed Ali ◽  
Daiki Mita ◽  
Wilfred Espulgar ◽  
Masato Saito ◽  
Masayuki Nishide ◽  
...  
Keyword(s):  
Single Cell ◽  
High Throughput Screening ◽  
Single Cell Analysis ◽  
Imaging System ◽  
Minimum Cost ◽  
Great Sensitivity ◽  
Monitoring Method ◽  
Cell Monitoring ◽  
Single Cell Isolation ◽  
Significant Release

A simple microengraving cell monitoring method for neutrophil extracellular traps (NETs) released from single neutrophils has been realized using a polydimethylsiloxane (PDMS) microwell array (MWA) sheet on a plasmon chip platform. An imbalance between NETs formation and the succeeding degradation (NETosis) are considered associated with autoimmune disease and its pathogenesis. Thus, an alternative platform that can conduct monitoring of this activity on single cell level at minimum cost but with great sensitivity is greatly desired. The developed MWA plasmon chips allow single cell isolation of neutrophils from 150 µL suspension (6.0 × 105 cells/mL) with an efficiency of 36.3%; 105 microwells with single cell condition. To demonstrate the utility of the chip, trapped cells were incubated between 2 to 4 h after introducing with 100 nM phorbol 12-myristate 13-acetate (PMA) before measurement. Under observation using a hyperspectral imaging system that allows high-throughput screening, the neutrophils stimulated by PMA solution show a significant release of fibrils and NETs after 4 h, with observed maximum areas between 314–758 µm2. An average absorption peak wavelength shows a redshift of Δλ = 1.5 nm as neutrophils release NETs.

scholarly journals Advances in Single-Cell Printing

Micromachines ◽  
2022 ◽  
Vol 13 (1) ◽  
pp. 80
Author(s):  
Xiaohu Zhou ◽  
Han Wu ◽  
Haotian Wen ◽  
Bo Zheng
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Three Dimensional ◽  
Cell Isolation ◽  
Special Focus ◽  
Cell Analysis ◽  
Cell Array ◽  
Cell Printing ◽  
Recent Developments ◽  
Single Cell Isolation

Single-cell analysis is becoming an indispensable tool in modern biological and medical research. Single-cell isolation is the key step for single-cell analysis. Single-cell printing shows several distinct advantages among the single-cell isolation techniques, such as precise deposition, high encapsulation efficiency, and easy recovery. Therefore, recent developments in single-cell printing have attracted extensive attention. We review herein the recently developed bioprinting strategies with single-cell resolution, with a special focus on inkjet-like single-cell printing. First, we discuss the common cell printing strategies and introduce several typical and advanced printing strategies. Then, we introduce several typical applications based on single-cell printing, from single-cell array screening and mass spectrometry-based single-cell analysis to three-dimensional tissue formation. In the last part, we discuss the pros and cons of the single-cell strategies and provide a brief outlook for single-cell printing.

scholarly journals Methods and sensors for functional genomic studies of cell-cycle transitions in single cells

2020 ◽  
Vol 52 (10) ◽  
pp. 468-477
Author(s):  
Alexander C. Zambon ◽  
Tom Hsu ◽  
Seunghee Erin Kim ◽  
Miranda Klinck ◽  
Jennifer Stowe ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Single Cell ◽  
Single Cell Analysis ◽  
Imaging System ◽  
Single Cells ◽  
Cell Types ◽  
Cell Analysis ◽  
Mcf7 Cells ◽  
Genomic Studies ◽  
Cell Subpopulations

Much of our understanding of the regulatory mechanisms governing the cell cycle in mammals has relied heavily on methods that measure the aggregate state of a population of cells. While instrumental in shaping our current understanding of cell proliferation, these approaches mask the genetic signatures of rare subpopulations such as quiescent (G0) and very slowly dividing (SD) cells. Results described in this study and those of others using single-cell analysis reveal that even in clonally derived immortalized cancer cells, ∼1–5% of cells can exhibit G0 and SD phenotypes. Therefore to enable the study of these rare cell phenotypes we established an integrated molecular, computational, and imaging approach to track, isolate, and genetically perturb single cells as they proliferate. A genetically encoded cell-cycle reporter (K67p-FUCCI) was used to track single cells as they traversed the cell cycle. A set of R-scripts were written to quantify K67p-FUCCI over time. To enable the further study G0 and SD phenotypes, we retrofitted a live cell imaging system with a micromanipulator to enable single-cell targeting for functional validation studies. Single-cell analysis revealed HT1080 and MCF7 cells had a doubling time of ∼24 and ∼48 h, respectively, with high duration variability in G1 and G2 phases. Direct single-cell microinjection of mRNA encoding (GFP) achieves detectable GFP fluorescence within ∼5 h in both cell types. These findings coupled with the possibility of targeting several hundreds of single cells improves throughput and sensitivity over conventional methods to study rare cell subpopulations.

scholarly journals Single-cell analysis of intestinal immune cells during helminth infection

2019 ◽  
Author(s):  
Laura Ferrer-Font ◽  
Palak Mehta ◽  
Phoebe Harmos ◽  
Alfonso Schmidt ◽  
Kylie M Price ◽  
...  
Keyword(s):  
Immune Responses ◽  
Single Cell ◽  
Immune Cells ◽  
Helminth Infection ◽  
Immune Cell ◽  
Single Cell Analysis ◽  
Cell Analysis ◽  
Mucus Production ◽  
Single Cell Isolation

AbstractSingle cell isolation from helminth infected intestines has been notoriously difficult, due to the strong anti-parasite type 2 immune responses that drive mucus production, tissue remodeling and immune cell infiltration. Through the systematic optimization of a standard intestinal digestion protocol, we were able to isolate millions of immune cells from heavily infected tissues. Using this protocol, we validated many hallmarks of anti-parasite immunity and analyzed immune cells from the lamina propria and granulomas during helminth development, as well as acute and chronic worm infection.

scholarly journals Cyto-Mine: An Integrated, Picodroplet System for High-Throughput Single-Cell Analysis, Sorting, Dispensing, and Monoclonality Assurance

2020 ◽  
Vol 25 (2) ◽  
pp. 177-189 ◽  
Author(s):  
Dimitris Josephides ◽  
Serena Davoli ◽  
William Whitley ◽  
Raphael Ruis ◽  
Robert Salter ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Line ◽  
High Throughput ◽  
Cell Sorting ◽  
High Throughput Screening ◽  
Single Cell Analysis ◽  
Microtiter Plate ◽  
Cell Analysis ◽  
Fully Integrated ◽  
Clonal Cell Lines

The primary goal of bioprocess cell line development is to obtain high product yields from robustly growing and well-defined clonal cell lines in timelines measured in weeks rather than months. Likewise, high-throughput screening of B cells and hybridomas is required for most cell line engineering workflows. A substantial bottleneck in these processes is detecting and isolating rare clonal cells with the required characteristics. Traditionally, this was achieved by the resource-intensive method of limiting dilution cloning, and more recently aided by semiautomated technologies such as cell sorting (e.g., fluorescence-activated cell sorting) and colony picking. In this paper we report on our novel Cyto-Mine Single Cell Analysis and Monoclonality Assurance System, which overcomes the limitations of current technologies by screening hundreds of thousands of individual cells for secreted target proteins, and then isolating and dispensing the highest producers into microtiter plate wells (MTP). The Cyto-Mine system performs this workflow using a fully integrated, microfluidic Cyto-Cartridge. Critically, all reagents and Cyto-Cartridges used are animal component-free (ACF) and sterile, thus allowing fast, robust, and safe isolation of desired cells.

Author response for "Immunocyte single cell analysis of vaccine‐induced narcolepsy"

2020 ◽  
Author(s):  
Alexander Lind ◽  
Falastin Salami ◽  
Anne‐Marie Landtblom ◽  
Lars Palm ◽  
Åke Lernmark ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cell Analysis ◽  
Author Response ◽  
Cell Analysis
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