Toward an understanding of immune cell sociology: real-time monitoring of cytokine secretion at the single-cell level

Cytokine secretion researches have been a main focus of studies among the scientists in the recent decades for its outstanding contribution to clinical diagnostics. Localized surface plasmon resonance (LSPR) technology is one of the conventional methods utilized to analyze these issues, as it could provide fast, label-free and real-time monitoring of biomolecule binding events. However, numerous LSPR-based biosensors in the past are usually utilized to monitor the average performance of cell groups rather than single cells. Meanwhile, the complicated sensor structures will lead to the fabrication and economic budget problems. Thus, in this paper, we report a simple synergistic integration of the cell trapping of microwell chip and gold-capped nanopillar-structured cyclo-olefin-polymer (COP) film for single cell level Interleukin 6 (IL-6) detection. Here, in-situ cytokine secreted from the trapped cell can be directly observed and analyzed through the peak red-shift in the transmittance spectrum. The fabricated device also shows the potential to conduct the real-time monitoring which would greatly help us identify the viability and biological variation of the tested single cell.

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Method for real-time monitoring of protein degradation at the single cell level

BioTechniques ◽

10.2144/000112453 ◽

2007 ◽

Vol 42 (4) ◽

pp. 446-450 ◽

Cited By ~ 53

Author(s):

Lijuan Zhang ◽

Nadya G. Gurskaya ◽

Ekaterina M. Merzlyak ◽

Dmitry B. Staroverov ◽

Nikolay N. Mudrik ◽

...

Keyword(s):

Protein Degradation ◽

Single Cell ◽

Real Time ◽

Single Cell Level ◽

Real Time Monitoring ◽

Cell Level

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Inkjet Printing-assisted Single-Cell Microarray on a Hydrophobic Surface Chip for Real-Time Monitoring of Enzyme Kinetics at Single-cell Level

Talanta ◽

10.1016/j.talanta.2020.122019 ◽

2021 ◽

pp. 122019

Author(s):

Xuexia Lin ◽

Feixiang Fang ◽

Chenjing Wang ◽

Ranjith Kumar Kankala ◽

Shufeng Zhou

Keyword(s):

Enzyme Kinetics ◽

Single Cell ◽

Real Time ◽

Inkjet Printing ◽

Hydrophobic Surface ◽

Single Cell Level ◽

Real Time Monitoring ◽

Cell Level ◽

Cell Microarray

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Controlling gas/liquid exchange using microfluidics for real-time monitoring of flagellar length in living Chlamydomonas at the single-cell level

Lab on a Chip ◽

10.1039/c2lc40638a ◽

2012 ◽

Vol 12 (21) ◽

pp. 4516 ◽

Cited By ~ 13

Author(s):

Xiaoni Ai ◽

Qionglin Liang ◽

Minna Luo ◽

Kai Zhang ◽

Junmin Pan ◽

...

Keyword(s):

Single Cell ◽

Real Time ◽

Single Cell Level ◽

Real Time Monitoring ◽

Cell Level

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3P105 Real-time Monitoring of Membrane Receptor Internalization : Low-Invasive, Quantitative and Single-Cell Level Measurement by SECM(03. Membrane proteins,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

Seibutsu Butsuri ◽

10.2142/biophys.54.s266_3 ◽

2014 ◽

Vol 54 (supplement1-2) ◽

pp. S266

Author(s):

Yoshiharu Matsumae ◽

Yasufumi Takahashi ◽

Kosuke Ino ◽

Hitoshi Shiku ◽

Tomokazu Matsue

Keyword(s):

Membrane Proteins ◽

Single Cell ◽

Real Time ◽

Annual Meeting ◽

Receptor Internalization ◽

Single Cell Level ◽

Real Time Monitoring ◽

Cell Level ◽

Level Measurement ◽

Biophysical Society

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Advanced Genomics-Based Approaches for Defining Allograft Rejection With Single Cell Resolution

Frontiers in Immunology ◽

10.3389/fimmu.2021.750754 ◽

2021 ◽

Vol 12 ◽

Author(s):

Tiffany Shi ◽

Krishna Roskin ◽

Brian M. Baker ◽

E. Steve Woodle ◽

David Hildeman

Keyword(s):

Single Cell ◽

Allograft Rejection ◽

Immune Cell ◽

Transplant Rejection ◽

Solid Organ Transplantation ◽

Genomic Analysis ◽

Solid Organ ◽

Single Cell Level ◽

Cell Level ◽

Network Analyses

Solid organ transplant recipients require long-term immunosuppression for prevention of rejection. Calcineurin inhibitor (CNI)-based immunosuppressive regimens have remained the primary means for immunosuppression for four decades now, yet little is known about their effects on graft resident and infiltrating immune cell populations. Similarly, the understanding of rejection biology under specific types of immunosuppression remains to be defined. Furthermore, development of innovative, rationally designed targeted therapeutics for mitigating or preventing rejection requires a fundamental understanding of the immunobiology that underlies the rejection process. The established use of microarray technologies in transplantation has provided great insight into gene transcripts associated with allograft rejection but does not characterize rejection on a single cell level. Therefore, the development of novel genomics tools, such as single cell sequencing techniques, combined with powerful bioinformatics approaches, has enabled characterization of immune processes at the single cell level. This can provide profound insights into the rejection process, including identification of resident and infiltrating cell transcriptomes, cell-cell interactions, and T cell receptor α/β repertoires. In this review, we discuss genomic analysis techniques, including microarray, bulk RNAseq (bulkSeq), single-cell RNAseq (scRNAseq), and spatial transcriptomic (ST) techniques, including considerations of their benefits and limitations. Further, other techniques, such as chromatin analysis via assay for transposase-accessible chromatin sequencing (ATACseq), bioinformatic regulatory network analyses, and protein-based approaches are also examined. Application of these tools will play a crucial role in redefining transplant rejection with single cell resolution and likely aid in the development of future immunomodulatory therapies in solid organ transplantation.

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Real-time quantification of protein expression at the single-cell level via dynamic protein synthesis translocation reporters

Nature Communications ◽

10.1038/ncomms11304 ◽

2016 ◽

Vol 7 (1) ◽

Cited By ~ 36

Author(s):

Delphine Aymoz ◽

Victoria Wosika ◽

Eric Durandau ◽

Serge Pelet

Keyword(s):

Protein Synthesis ◽

Protein Expression ◽

Single Cell ◽

Real Time ◽

Single Cell Level ◽

Cell Level

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Measurement of Molecular Mobility in Mammalian Cells

Advances in Bioengineering ◽

10.1115/imece2004-61508 ◽

2004 ◽

Author(s):

Alptekin Aksan ◽

Mehmet Toner

Keyword(s):

Single Cell ◽

Real Time ◽

Chemical Reaction ◽

Molecular Mobility ◽

Mammalian Cells ◽

Free Water ◽

Single Cell Level ◽

Extracellular Water ◽

Cell Level

Preservation of mammalian cells requires establishing a reversible stasis condition by reducing the intra/extracellular molecular mobility ensuring reduced chemical reaction and deterioration rates. Molecular mobility may be reduced by various techniques. For example, in cryopreservation, mobility within and surrounding the cell is reduced through freezing the free water that constitutes 70–90% of the cell’s composition. In dried-state preservation applied successfully to preserve seeds, pharmacological materials and foodstuff (mimicking the anhydrobiosis phenomenon seen in nature), reduction in molecular mobility is established by removing intra/extracellular water. Certain carbohydrates (such as trehalose and sucrose) can be artificially uploaded into mammalian cells to replace the removed water and to form an intra/extracellular glass. In this research, a fluorescent rotor is utilized to determine the changes in intracellular molecular mobility during carbohydrate uploading of mammalian cells. It was shown that using this technique, it is feasible to make real-time mobility measurements at a single cell level.

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