Single-cell PCR of genomic DNA enabled by automated single-cell printing for cell isolation

2015 ◽  
Vol 69 ◽  
pp. 301-306 ◽  
Author(s):  
F. Stumpf ◽  
J. Schoendube ◽  
A. Gross ◽  
C. Rath ◽  
S. Niekrawietz ◽  
...  
Keyword(s):  
Single Cell ◽  
Genomic Dna ◽  
Cell Isolation ◽  
Cell Printing ◽  
Single Cell Pcr

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 THE PHYLOGENETIC TREE OF Alexandrium Prorocentrum AND PSEUDO-NITZSCHIA OF HARMFUL AND TOXIC ALGAE IN VIETNAM COASTAL WATERS BASED ON SEQUENCES OF 18sRdna, Its1-5.8s-Its2 GENE FRAGMENTS AND SINGLE CELL -PER METHOD

1970 ◽  
Vol 32 (2) ◽  
pp. 203-218
Author(s):  
Dang Diem Hong ◽  
Ngo Thi Hoai Thu ◽  
Hoang Sy Nam ◽  
Hoang Minh Hien ◽  
Luyen Quoc Hai ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Nucleotide Sequence ◽  
Single Cell ◽  
Coastal Waters ◽  
18S Rdna ◽  
Genomic Dna ◽  
Nucleotide Sequence Analysis ◽  
Single Cell Pcr ◽  
Pcr Method ◽  
Molecular Biological Techniques

Molecular biological techniques support the identification of microalgae of Vietnam. Prorocentrum, Alexandrium and Pseudo-nitzschia are main harmful and toxic microalgal genera found in Vietnam coastal waters. The results of morphology and nucleotide sequence analysis of 18S rDNA and ITS 1-5.8S-ITS2 gene obtained from genomic DNA have shown that the Prorocentrum sp. 3 (isolated from Cat Ba, Hai Phong on October, 2004), Alexandrium sp. 5 (collected on October, 2004) and Pseudo-nitzschia sp. G3 (colleted in Do Son, Hai Phong on December, 2005) belonging to Prorocentrum mexicanum (the homological percent of 99.9% with sequencing of P mexicanum in Genbank ofY16232, AY886763), A. minutum (99.8% - AJ535388, DQ168664) and Pseudo-nitzschia pungens (98.8% - AY544769, DQ166533), respectively. The obtained results indicated that the exceptional fresh samples, for Prorocentrum genus which may be preserved at 25% ethanol, 4% formaldehyde, 1% glutaraldehyde, while Alexandrium genus ó at 25% ethanol, and Pseudo-nitzschia genus - 4% formaldehyde and 1% glutaraldehyde for two weeks have not effected on their analysis of sequences generated by Single Cell PCR method. In these studied samples, the nucleotide sequences obtained from genomic DNA and Single-cell PCR methods were the same with the homological percent more than 99%. Application of this method to samples collected from Phu Quoc Island, southern part of Vietnam, in 27-29, June, 2006 showed that Prorocentrum mican and P sigmoisdes were found.

A microfluidic device enabling deterministic single cell trapping and release

Lab on a Chip ◽  
2021 ◽  
Author(s):  
Huichao Chai ◽  
Yongxiang Feng ◽  
Fei Liang ◽  
Wenhui Wang
Keyword(s):  
Chemical Analysis ◽  
Single Cell ◽  
Microfluidic Device ◽  
Single Cells ◽  
Cell Isolation ◽  
Cell Trapping ◽  
Analysis Techniques ◽  
Single Cell Trapping ◽  
Single Cell Isolation ◽  
Made In

Successful single-cell isolation is a pivotal technique for subsequent biological and chemical analysis of single cells. Although significant advances have been made in single-cell isolation and analysis techniques, most passive...

Application Of Isoraft Single Cell Isolation For Analysis Of Pediatric Bal Macrophages

2014 ◽  
Vol 133 (2) ◽  
pp. AB142
Author(s):  
Neil Alexis ◽  
Heather Wells ◽  
Yogesh Saini ◽  
Louisa Brighton ◽  
Nancy Allbritton ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Isolation ◽  
Single Cell Isolation

Grids from bands, or bands from grids? An examination of the effects of single unit contamination on grid cell firing fields

2016 ◽  
Vol 115 (2) ◽  
pp. 992-1002 ◽  
Author(s):  
Z. Navratilova ◽  
K. B. Godfrey ◽  
B. L. McNaughton
Keyword(s):  
Single Cell ◽  
Grid Cell ◽  
Cell Isolation ◽  
Recording Technology ◽  
Firing Patterns ◽  
Limiting Step ◽  
Isolation Methods ◽  
Single Cell Isolation ◽  
Cell Firing ◽  
Cell Data

Neural recording technology is improving rapidly, allowing for the detection of spikes from hundreds of cells simultaneously. The limiting step in multielectrode electrophysiology continues to be single cell isolation. However, this step is crucial to the interpretation of data from putative single neurons. We present here, in simulation, an illustration of possibly erroneous conclusions that may be reached when poorly isolated single cell data are analyzed. Grid cells are neurons recorded in rodents, and bats, that spike in equally spaced locations in a hexagonal pattern. One theory states that grid firing patterns arise from a combination of band firing patterns. However, we show here that summing the grid firing patterns of two poorly resolved neurons can result in spurious band-like patterns. Thus, evidence of neurons spiking in band patterns must undergo extreme scrutiny before it is accepted. Toward this aim, we discuss single cell isolation methods and metrics.

Clonal Isolation of Hybridomas by Manual Single-Cell Isolation

Hybridoma ◽  
1983 ◽  
Vol 2 (2) ◽  
pp. 231-234 ◽  
Author(s):  
R.J. SIJENS ◽  
A.A.M. THOMAS ◽  
A. JACKERS ◽  
A. BOEYÉ
Keyword(s):  
Single Cell ◽  
Cell Isolation ◽  
Single Cell Isolation

scholarly journals Characterization of CRISPR/Cas9 RANKL knockout mesenchymal stem cell clones based on single-cell printing technology and emulsion coupling assay as a low-cellularity workflow for single-cell cloning

2020 ◽  
Author(s):  
Tobias Groß ◽  
Csaba Jeney ◽  
Darius Halm ◽  
Günter Finkenzeller ◽  
G. Björn Stark ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Line ◽  
Large Scale ◽  
Single Cells ◽  
Protein Detection ◽  
Cell Line Development ◽  
Cell Printing ◽  
Cell Clones ◽  
The University

AbstractThe homogeneity of the genetically modified single-cells is a necessity for many applications such as cell line development, gene therapy, and tissue engineering and in particular for regenerative medical applications. The lack of tools to effectively isolate and characterize CRISPR/Cas9 engineered cells is considered as a significant bottleneck in these applications. Especially the incompatibility of protein detection technologies to confirm protein expression changes without a preconditional large-scale clonal expansion, creates a gridlock in many applications. To ameliorate the characterization of engineered cells, we propose an improved workflow, including single-cell printing/isolation technology based on fluorescent properties with high yield, a genomic edit screen (surveyor assay), mRNA rtPCR assessing altered gene expression and a versatile protein detection tool called emulsion-coupling to deliver a high-content, unified single-cell workflow. The workflow was exemplified by engineering and functionally validating RANKL knockout immortalized mesenchymal stem cells showing altered bone formation capacity of these cells. The resulting workflow is economical, without the requirement of large-scale clonal expansions of the cells with overall cloning efficiency above 30% of CRISPR/Cas9 edited cells. Nevertheless, as the single-cell clones are comprehensively characterized at an early, highly parallel phase of the development of cells including DNA, RNA, and protein levels, the workflow delivers a higher number of successfully edited cells for further characterization, lowering the chance of late failures in the development process.Author summaryI completed my undergraduate degree in biochemistry at the University of Ulm and finished my master's degree in pharmaceutical biotechnology at the University of Ulm and University of applied science of Biberach with a focus on biotechnology, toxicology and molecular biology. For my master thesis, I went to the University of Freiburg to the department of microsystems engineering, where I developed a novel workflow for cell line development. I stayed at the institute for my doctorate, but changed my scientific focus to the development of the emulsion coupling technology, which is a powerful tool for the quantitative and highly parallel measurement of protein and protein interactions. I am generally interested in being involved in the development of innovative molecular biological methods that can be used to gain new insights about biological issues. I am particularly curious to unravel the complex and often poorly understood protein interaction pathways that are the cornerstone of understanding cellular functionality and are a fundamental necessity to describe life mechanistically.

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