Single Cell Interrogation using Optofluidic Platforms for Systems Immunology

MRS Advances ◽  
2016 ◽  
Vol 1 (56) ◽  
pp. 3783-3788 ◽  
Author(s):  
Serap Aksu
Keyword(s):  
Immune System ◽  
Single Cell ◽  
High Throughput ◽  
Individual Cell ◽  
Cytokine Secretion ◽  
Time Dependent ◽  
Label Free ◽  
System Behavior ◽  
Systems Immunology

ABSTRACTThe main objective of this report is to demonstrate novel engineering technologies to investigate regulatory mechanisms of systems immunology in a time-dependent and high-throughput manner. Understanding of immune system behavior is crucial for accurate prognosis of infections and identification of diseases at early stage. An ultimate goal of biomedical engineering is to develop predictive models of immune system behavior in tissue, which necessitates a comprehensive map of dynamic (time-dependent) input-output relationships at the individual cell level. Traditionally, biochemical analysis on the cell signaling is obtained from bulky cell ensembles which average over relevant individual cell response. The response consists firstly of signaling protein (cytokine) secretions which are released during a disease state and which are used to activate the immune system to respond to the disease. We investigate the cytokine secretion dynamics of a single immune cell in response to the stimulant using automated and comprehensive optofluidic platforms. These platforms enable survival and manipulation of single cells in compartments having compatible sizes with cells as well as provide precise control over the type, dose and time-course of the stimulant. The cytokine secretion dynamics of single cell are typically explained by measuring the types, rates, frequencies and concentrations of various cytokines. For the quantitative measurements, label free localized surface plasmon resonance (LSPR) based biosensor can be integrated within the microfluidic device. Microfluidic channels can confine secreted cytokines in compartments, minimize dilution effects and increase detection sensitivity for label free plasmonic biosensing. The direct application of LSPR to in-situ live cell function analysis is still in its infancy and use of such in-situ, real time, and label free biodetection will effortlessly provide high-throughput quantitative bioanalysis for understanding immune system behavior.

scholarly journals Label-Free and Quantitative Dry Mass Monitoring for Single Cells during In Situ Culture

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1635
Author(s):  
Ya Su ◽  
Rongxin Fu ◽  
Wenli Du ◽  
Han Yang ◽  
Li Ma ◽  
...  
Keyword(s):  
Cell Growth ◽  
Single Cell ◽  
Hela Cells ◽  
Quantitative Measurement ◽  
Single Cells ◽  
Dry Mass ◽  
Quantitative Detection ◽  
Label Free ◽  
Mass Sensitivity

Quantitative measurement of single cells can provide in-depth information about cell morphology and metabolism. However, current live-cell imaging techniques have a lack of quantitative detection ability. Herein, we proposed a label-free and quantitative multichannel wide-field interferometric imaging (MWII) technique with femtogram dry mass sensitivity to monitor single-cell metabolism long-term in situ culture. We demonstrated that MWII could reveal the intrinsic status of cells despite fluctuating culture conditions with 3.48 nm optical path difference sensitivity, 0.97 fg dry mass sensitivity and 2.4% average maximum relative change (maximum change/average) in dry mass. Utilizing the MWII system, different intrinsic cell growth characteristics of dry mass between HeLa cells and Human Cervical Epithelial Cells (HCerEpiC) were studied. The dry mass of HeLa cells consistently increased before the M phase, whereas that of HCerEpiC increased and then decreased. The maximum growth rate of HeLa cells was 11.7% higher than that of HCerEpiC. Furthermore, HeLa cells were treated with Gemcitabine to reveal the relationship between single-cell heterogeneity and chemotherapeutic efficacy. The results show that cells with higher nuclear dry mass and nuclear density standard deviations were more likely to survive the chemotherapy. In conclusion, MWII was presented as a technique for single-cell dry mass quantitative measurement, which had significant potential applications for cell growth dynamics research, cell subtype analysis, cell health characterization, medication guidance and adjuvant drug development.

scholarly journals Integration of an In Situ MALDI-Based High-Throughput Screening Process: A Case Study with Receptor Tyrosine Kinase c-MET

2017 ◽  
Vol 22 (10) ◽  
pp. 1203-1210 ◽  
Author(s):  
Katrin Beeman ◽  
Jens Baumgärtner ◽  
Manuel Laubenheimer ◽  
Karlheinz Hergesell ◽  
Martin Hoffmann ◽  
...  
Keyword(s):  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Maldi Ms ◽  
Kinase Assay ◽  
Label Free ◽  
Screening Process ◽  
Label Free Detection ◽  
Ic50 Values

Mass spectrometry (MS) is known for its label-free detection of substrates and products from a variety of enzyme reactions. Recent hardware improvements have increased interest in the use of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS for high-throughput drug discovery. Despite interest in this technology, several challenges remain and must be overcome before MALDI-MS can be integrated as an automated “in-line reader” for high-throughput drug discovery. Two such hurdles include in situ sample processing and deposition, as well as integration of MALDI-MS for enzymatic screening assays that usually contain high levels of MS-incompatible components. Here we adapt our c-MET kinase assay to optimize for MALDI-MS compatibility and test its feasibility for compound screening. The pros and cons of the Echo (Labcyte) as a transfer system for in situ MALDI-MS sample preparation are discussed. We demonstrate that this method generates robust data in a 1536-grid format. We use the MALDI-MS to directly measure the ratio of c-MET substrate and phosphorylated product to acquire IC50 curves and demonstrate that the pharmacology is unaffected. The resulting IC50 values correlate well between the common label-based capillary electrophoresis and the label-free MALDI-MS detection method. We predict that label-free MALDI-MS-based high-throughput screening will become increasingly important and more widely used for drug discovery.

scholarly journals High-throughput, label-free, single-cell photoacoustic microscopy of intratumoral metabolic heterogeneity

2019 ◽  
Vol 3 (5) ◽  
pp. 381-391 ◽  
Author(s):  
Pengfei Hai ◽  
Toru Imai ◽  
Song Xu ◽  
Ruiying Zhang ◽  
Rebecca L. Aft ◽  
...  
Keyword(s):  
Single Cell ◽  
High Throughput ◽  
Photoacoustic Microscopy ◽  
Label Free ◽  
Metabolic Heterogeneity

Marker-free detection of progenitor cell differentiation by analysis of Brownian motion in micro-wells

2015 ◽  
Vol 7 (2) ◽  
pp. 178-183 ◽  
Author(s):  
Farzad Sekhavati ◽  
Max Endele ◽  
Susanne Rappl ◽  
Anna-Kristina Marel ◽  
Timm Schroeder ◽  
...  
Keyword(s):  
Brownian Motion ◽  
Cell Differentiation ◽  
Single Cell ◽  
High Throughput ◽  
Progenitor Cell ◽  
Single Cell Level ◽  
Label Free ◽  
Cell Level ◽  
Marker Free

The analysis of Brownian motion is a sensitive and robust tool for a label-free high-throughput investigation of cell differentiation at the single-cell level.

A high-throughput flow cytometry-on-a-CMOS platform for single-cell dielectric spectroscopy at microwave frequencies

Lab on a Chip ◽  
2018 ◽  
Vol 18 (14) ◽  
pp. 2065-2076 ◽  
Author(s):  
Jun-Chau Chien ◽  
Ali Ameri ◽  
Erh-Chia Yeh ◽  
Alison N. Killilea ◽  
Mekhail Anwar ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Dielectric Properties ◽  
Single Cell ◽  
High Throughput ◽  
Dielectric Spectroscopy ◽  
Free Flow ◽  
Label Free ◽  
Microwave Frequencies

This work presents a microfluidics-integrated label-free flow cytometry-on-a-CMOS platform for the characterization of the cytoplasm dielectric properties at microwave frequencies.

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