Time-resolved ICP-MS analysis of mineral element contents and distribution patterns in single cells

The Analyst ◽  
2015 ◽  
Vol 140 (2) ◽  
pp. 523-531 ◽  
Author(s):  
Hailong Wang ◽  
Bing Wang ◽  
Meng Wang ◽  
Lingna Zheng ◽  
Hanqing Chen ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cells ◽  
Distribution Patterns ◽  
Clinical Applications ◽  
Mineral Element ◽  
Time Resolved ◽  
Ms Analysis ◽  
Icp Ms ◽  
Element Contents

Novel single cell techniques are attracting growing interest for clinical applications, because they can elucidate the cellular diversity and heterogeneity instead of the average masked by bulk measurements.

Highly efficient single-cell analysis of microbial cells by time-resolved inductively coupled plasma mass spectrometry

2014 ◽  
Vol 29 (9) ◽  
pp. 1598-1606 ◽  
Author(s):  
Shin-ichi Miyashita ◽  
Alexander S. Groombridge ◽  
Shin-ichiro Fujii ◽  
Ayumi Minoda ◽  
Akiko Takatsu ◽  
...  
Keyword(s):  
Single Cell ◽  
Inductively Coupled Plasma ◽  
Single Cell Analysis ◽  
High Time Resolution ◽  
Time Resolved ◽  
Microbial Cells ◽  
Inductively Coupled ◽  
Highly Efficient ◽  
Icp Ms ◽  
Plasma Mass Spectrometry

Highly efficient single-cell elemental analysis of microbial cells was achieved using a developed ICP-MS system with approximately 100% cell introduction efficiency and high time resolution.

scholarly journals Massively parallel and time-resolved RNA sequencing in single cells with scNT-Seq

2021 ◽  
Author(s):  
Qi Qiu ◽  
Peng Hu ◽  
Hao Wu
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Temporal Dynamics ◽  
Single Cells ◽  
Chemical Conversion ◽  
Droplet Microfluidics ◽  
Massively Parallel ◽  
One Pot ◽  
Time Resolved ◽  
Rna Biogenesis

Abstract Single-cell RNA sequencing offers snapshots of whole transcriptomes but obscures the temporal dynamics of RNA biogenesis and decay. Here we present single-cell metabolically labeled new RNA tagging sequencing (scNT-Seq), a method for massively parallel analysis of newly-transcribed and pre-existing RNAs from the same cell. This droplet microfluidics-based method enables high-throughput chemical conversion on barcoded beads, efficiently marking newly-transcribed RNAs with T-to-C substitutions. The steps of the protocol are (1) metabolically labeling of cells with 4sU, (2) co-encapsulating individual cell with a barcoded oligo-dT primer coated bead in a nanoliter-scale droplet, (3) performing one-pot 4sU chemical conversion on pooled barcoded beads, and (4) reverse transcription, cDNA amplification, tagmentation, indexing PCR, and sequencing. scNT-Seq provides a broadly applicable strategy to investigate dynamic biological systems at single-cell resolution.

Forensic comparison of PVC tape backings using time resolved LA-ICP-MS analysis

2019 ◽  
Vol 12 ◽  
pp. 33-41 ◽  
Author(s):  
Felix Kuczelinis ◽  
Peter Weis ◽  
Nicolas-H. Bings
Keyword(s):  
Time Resolved ◽  
Ms Analysis ◽  
Icp Ms

scholarly journals Quantification of single-cell nanoparticle concentrations and the distribution of these concentrations in cell population

2014 ◽  
Vol 11 (94) ◽  
pp. 20131152 ◽  
Author(s):  
Jason T. Rashkow ◽  
Sunny C. Patel ◽  
Ryan Tappero ◽  
Balaji Sitharaman
Keyword(s):  
Single Cell ◽  
Inductively Coupled Plasma ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Large Population ◽  
Cellular Heterogeneity ◽  
Physical Parameters ◽  
Animal Cells ◽  
Promising Tool ◽  
Icp Ms

Quantification of nanoparticle uptake into cells is necessary for numerous applications in cellular imaging and therapy. Herein, synchrotron X-ray fluorescence (SXRF) microscopy, a promising tool to quantify elements in plant and animal cells, was employed to quantify and characterize the distribution of titanium dioxide (TiO 2 ) nanosphere uptake in a population of single cells. These results were compared with average nanoparticle concentrations per cell obtained by widely used inductively coupled plasma mass spectrometry (ICP-MS). The results show that nanoparticle concentrations per cell quantified by SXRF were of one to two orders of magnitude greater compared with ICP-MS. The SXRF results also indicate a Gaussian distribution of the nanoparticle concentration per cell. The results suggest that issues relevant to the field of single-cell analysis, the limitation of methods to determine physical parameters from large population averages leading to potentially misleading information and the lack of any information about the cellular heterogeneity are equally relevant for quantification of nanoparticles in cell populations.

Interrogating the variation of element masses and distribution patterns in single cells using ICP-MS with a high efficiency cell introduction system

2016 ◽  
Vol 409 (5) ◽  
pp. 1415-1423 ◽  
Author(s):  
Hailong Wang ◽  
Meng Wang ◽  
Bing Wang ◽  
Lingna Zheng ◽  
Hanqing Chen ◽  
...  
Keyword(s):  
High Efficiency ◽  
Single Cells ◽  
Distribution Patterns ◽  
Icp Ms

scholarly journals Time-resolved assessment of single-cell protein secretion by sequencing

2021 ◽  
Author(s):  
Tongjin Wu ◽  
Howard John Womersley ◽  
Jiehao Wang ◽  
Jonathan Adam Scolnick ◽  
Lih Feng Cheow
Keyword(s):  
Single Cell ◽  
Protein Secretion ◽  
Single Cells ◽  
Transcript Abundance ◽  
Cellular Protein ◽  
Single Cell Protein ◽  
Cytokine Secretion ◽  
Cell Protein ◽  
Time Resolved ◽  
Tnf Α

Secreted proteins play critical roles in cellular communication and functional orchestration. Methods enabling concurrent measurement of cellular protein secretion, phenotypes and transcriptomes are still unavailable. Here, we describe time-resolved assessment of protein secretion from single cells by sequencing (TRAPS-seq). Released proteins are trapped onto cell surface via affinity matrices, and the captured analytes together with phenotypic markers can be probed by oligonucleotide-barcoded antibodies and simultaneously sequenced with transcriptomes. We used TRAPS-seq to interrogate secretion dynamics of pleiotropic cytokines (IFN-γ, IL-2 and TNF-α) of early activated human T lymphocytes, unraveling limited correlation between cytokine secretion and its transcript abundance with regard to timing and strength. We found that early central memory T cells with CD45RA expression (TCMRA) are the most effective responders in multiple cytokine secretion, and polyfunctionality involves unique yet dynamic combinations of gene signatures over time. TRAPS-seq presents a useful tool for cellular indexing of secretions, phenotypes, and transcriptomes at single-cell resolution.

scholarly journals Extraction of nuclei from archived post-mortem tissues for single-nucleus sequencing applications

2020 ◽  
Author(s):  
Malosree Maitra ◽  
Corina Nagy ◽  
Yu Chang Wang ◽  
Camila Nascimento ◽  
Matthew Suderman ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cells ◽  
Clinical Applications ◽  
Post Mortem ◽  
Rna Seq ◽  
High Quality ◽  
Limiting Step ◽  
Single Nucleus ◽  
Rna And Dna ◽  
Variable Quality

Abstract Single-cell and single-nucleus sequencing techniques are a burgeoning field with various biological, biomedical, and clinical applications. Numerous high and low-throughput methods have been developed for sequencing the RNA and DNA content of single cells. However, for all these methods the key requirement is high quality input of a single-cell or single-nucleus suspension. Preparing such a suspension is the limiting step when working with fragile, archived tissues of variable quality. This hurdle can prevent such tissues from being extensively investigated with single-cell technologies. We describe a protocol for preparing single-nucleus suspensions within the span of a few hours that reliably works for multiple post-mortem and archived tissues types using standard lab equipment. Moreover, these preparations are compatible with single-nucleus RNA-seq and ATAC-seq using the 10X Genomics’ Chromium system.

scholarly journals HyPR-seq: Single-cell quantification of chosen RNAs via hybridization and sequencing of DNA probes

2020 ◽  
Vol 117 (52) ◽  
pp. 33404-33413
Author(s):  
Jamie L. Marshall ◽  
Benjamin R. Doughty ◽  
Vidya Subramanian ◽  
Philine Guckelberger ◽  
Qingbo Wang ◽  
...  
Keyword(s):  
Single Cell ◽  
Single Cells ◽  
High Sensitivity ◽  
Dna Probes ◽  
Cellular Heterogeneity ◽  
Marker Genes ◽  
Time Resolved ◽  
Rna Profiling ◽  
Cell Quantification ◽  
Low Sensitivity

Single-cell quantification of RNAs is important for understanding cellular heterogeneity and gene regulation, yet current approaches suffer from low sensitivity for individual transcripts, limiting their utility for many applications. Here we present Hybridization of Probes to RNA for sequencing (HyPR-seq), a method to sensitively quantify the expression of hundreds of chosen genes in single cells. HyPR-seq involves hybridizing DNA probes to RNA, distributing cells into nanoliter droplets, amplifying the probes with PCR, and sequencing the amplicons to quantify the expression of chosen genes. HyPR-seq achieves high sensitivity for individual transcripts, detects nonpolyadenylated and low-abundance transcripts, and can profile more than 100,000 single cells. We demonstrate how HyPR-seq can profile the effects of CRISPR perturbations in pooled screens, detect time-resolved changes in gene expression via measurements of gene introns, and detect rare transcripts and quantify cell-type frequencies in tissue using low-abundance marker genes. By directing sequencing power to genes of interest and sensitively quantifying individual transcripts, HyPR-seq reduces costs by up to 100-fold compared to whole-transcriptome single-cell RNA-sequencing, making HyPR-seq a powerful method for targeted RNA profiling in single cells.

scholarly journals Time-resolved ICP-MS Measurement: a New Method for Elemental and Multiparametric Analysis of Single Cells

2014 ◽  
Vol 30 (2) ◽  
pp. 219-224 ◽  
Author(s):  
Shin-ichi MIYASHITA ◽  
Alexander S. GROOMBRIDGE ◽  
Shin-ichiro FUJII ◽  
Akiko TAKATSU ◽  
Koichi CHIBA ◽  
...  
Keyword(s):  
Single Cells ◽  
New Method ◽  
Time Resolved ◽  
Multiparametric Analysis ◽  
Icp Ms
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