Single-cell level point mutation analysis of circulating tumor cells through droplet microfluidics

2021 ◽  
Author(s):  
Shihui Qiu ◽  
Chuanjie Shen ◽  
Xiaoyu Jian ◽  
Yunxing Lu ◽  
Zhaoduo Tong ◽  
...  
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Point Mutation ◽  
Circulating Tumor Cells ◽  
Mutation Analysis ◽  
Droplet Microfluidics ◽  
Single Cell Level ◽  
Cell Level

scholarly journals Genomic Analysis of Circulating Tumor Cells at the Single-Cell Level

2020 ◽  
Vol 22 (6) ◽  
pp. 770-781 ◽  
Author(s):  
Shan Lu ◽  
Chia-Jung Chang ◽  
Yinghui Guan ◽  
Edith Szafer-Glusman ◽  
Elizabeth Punnoose ◽  
...  
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Genomic Analysis ◽  
Single Cell Level ◽  
Cell Level

A microfluidic platform for high-purity separating circulating tumor cells at the single-cell level

Talanta ◽  
2019 ◽  
Vol 200 ◽  
pp. 169-176 ◽  
Author(s):  
Kun Wang ◽  
Lin Zhou ◽  
Simin Zhao ◽  
Zule Cheng ◽  
Shihui Qiu ◽  
...  
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
High Purity ◽  
Single Cell Level ◽  
Cell Level ◽  
Microfluidic Platform

scholarly journals Heterogeneity of PIK3CA mutational status at the single cell level in circulating tumor cells from metastatic breast cancer patients

2014 ◽  
Vol 9 (4) ◽  
pp. 749-757 ◽  
Author(s):  
Marta Pestrin ◽  
Francesca Salvianti ◽  
Francesca Galardi ◽  
Francesca De Luca ◽  
Natalie Turner ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Metastatic Breast Cancer ◽  
Single Cell ◽  
Cancer Patients ◽  
Tumor Cells ◽  
Metastatic Breast ◽  
Single Cell Level ◽  
Breast Cancer Patients ◽  
Cell Level ◽  
Mutational Status

scholarly journals Combined Molecular Genetic and Cytogenetic Analysis from Single Cells after Isothermal Whole-Genome Amplification

2011 ◽  
Vol 57 (7) ◽  
pp. 1032-1041 ◽  
Author(s):  
Thomas Kroneis ◽  
Jochen B Geigl ◽  
Amin El-Heliebi ◽  
Martina Auer ◽  
Peter Ulz ◽  
...  
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Whole Genome Amplification ◽  
Single Cells ◽  
Molecular Genetic ◽  
Target Cells ◽  
Whole Genome ◽  
Single Cell Level ◽  
Cell Level ◽  
Genome Amplification

BACKGROUND Analysis of chromosomal aberrations or single-gene disorders from rare fetal cells circulating in the blood of pregnant women requires verification of the cells' genomic identity. We have developed a method enabling multiple analyses at the single-cell level that combines verification of the genomic identity of microchimeric cells with molecular genetic and cytogenetic diagnosis. METHODS We used a model system of peripheral blood mononuclear cells spiked with a colon adenocarcinoma cell line and immunofluorescence staining for cytokeratin in combination with DNA staining with the nuclear dye TO-PRO-3 in a preliminary study to define candidate microchimeric (tumor) cells in Cytospin preparations. After laser microdissection, we performed low-volume on-chip isothermal whole-genome amplification (iWGA) of single and pooled cells. RESULTS DNA fingerprint analysis of iWGA aliquots permitted successful identification of all analyzed candidate microchimeric cell preparations (6 samples of pooled cells, 7 samples of single cells). Sequencing of 3 single-nucleotide polymorphisms was successful at the single-cell level for 20 of 32 allelic loci. Metaphase comparative genomic hybridization (mCGH) with iWGA products of single cells showed the gains and losses known to be present in the genomic DNA of the target cells. CONCLUSIONS This method may be instrumental in cell-based noninvasive prenatal diagnosis. Furthermore, the possibility to perform mCGH with amplified DNA from single cells offers a perspective for the analysis of nonmicrochimeric rare cells exhibiting genomic alterations, such as circulating tumor cells.

scholarly journals Development of an optimal protocol for molecular profiling of tumor cells in pleural effusions at single‐cell level

2021 ◽  
Author(s):  
Ikuko Takeda Nakamura ◽  
Masachika Ikegami ◽  
Nobuhiko Hasegawa ◽  
Takuo Hayashi ◽  
Toshihide Ueno ◽  
...  
Keyword(s):  
Single Cell ◽  
Tumor Cells ◽  
Molecular Profiling ◽  
Single Cell Level ◽  
Pleural Effusions ◽  
Cell Level ◽  
Optimal Protocol

scholarly journals “Antibiotic inhibition of bacteria growth in droplets reveals heteroresistance pattern at the single cell level”

2018 ◽  
Author(s):  
Ott Scheler ◽  
Karol Makuch ◽  
Pawel R. Debski ◽  
Michal Horka ◽  
Artur Ruszczak ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Single Cell ◽  
Agar Plate ◽  
Droplet Microfluidics ◽  
Resistance Pattern ◽  
Single Cell Level ◽  
Cell Level ◽  
High Throughput Analysis ◽  
Bacteria Growth ◽  
Drug Resistance Profile

Heteroresistance is a phenomenon where isogenic bacteria population exhibits a diverse antibiotic resistance pattern at sub-population or single cell level. The sub-populations with higher resistance can remain undetected with conventional diagnostics which makes them subsequently harder to treat. Such surviving phenotypically heterogeneous sub-populations are also a potential hotbed for novel mutations, thus increasing the resistance permanently in bacteria. Droplet microfluidics gives tools for high-throughput analysis of bacteria and their response to antibiotics at single cell level, which is difficult to obtain with traditional agar plate technologies. In here we show for the first time the precise digital quantification of drug resistance profile in isogenic population at single cell level. We also see that the inhibiting amount of drug per bacteria remains quite stable regardless of bacteria density. Interestingly, the bacteria clump together preferably near these sub-inhibitory conditions. The technology and findings we describe here provide novel quantitative insight into the heteroresistance which is a key step in understanding the pathways leading to drug resistance. This knowledge is crucial in the context of global drug resistance threat as it can help us to find tools to prevent further escalation of drug resistance.

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