Advanced Optogenetic-Based Biosensing and Related Biomaterials

The ability to stimulate mammalian cells with light, brought along by optogenetic control, has significantly broadened our understanding of electrically excitable tissues. Backed by advanced (bio)materials, it has recently paved the way towards novel biosensing concepts supporting bio-analytics applications transversal to the main biomedical stream. The advancements concerning enabling biomaterials and related novel biosensing concepts involving optogenetics are reviewed with particular focus on the use of engineered cells for cell-based sensing platforms and the available toolbox (from mere actuators and reporters to novel multifunctional opto-chemogenetic tools) for optogenetic-enabled real-time cellular diagnostics and biosensor development. The key advantages of these modified cell-based biosensors concern both significantly faster (minutes instead of hours) and higher sensitivity detection of low concentrations of bioactive/toxic analytes (below the threshold concentrations in classical cellular sensors) as well as improved standardization as warranted by unified analytic platforms. These novel multimodal functional electro-optical label-free assays are reviewed among the key elements for optogenetic-based biosensing standardization. This focused review is a potential guide for materials researchers interested in biosensing based on light-responsive biomaterials and related analytic tools.

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Comparison of Leading Biosensor Technologies to Measure Endothelial Adhesion, Barrier Properties, and Responses to Cytokines in Real-Time

Proceedings ◽

10.3390/iecb2020-07036 ◽

2020 ◽

Vol 60 (1) ◽

pp. 62

Author(s):

James J. W. Hucklesby ◽

Akshata Anchan ◽

Simon J. O’Carroll ◽

Catherine E. Angel ◽

E. Scott Graham

Keyword(s):

Real Time ◽

Barrier Properties ◽

Microvascular Endothelial Cell ◽

Label Free ◽

Adhesion Barrier ◽

Impedance Sensing ◽

Non Invasive ◽

Reduced Frequency ◽

Cell Substrate ◽

Higher Sensitivity

Electric Cell-substrate Impedance Sensing (ECIS), xCELLigence and cellZscope are commercially available instruments which are able to measure the impedance of cellular monolayers continuously and with high precision. The small currents used allow the label-free, real-time monitoring of the cells in a non-invasive manner. Despite the widespread use of these systems individually, direct comparisons between the systems have not been published. In order to compare the sensitivity of the instruments, the responses of the brain microvascular endothelial cell line hCMVEC to the inflammatory cytokines TNFα and IL1β were measured on all three instruments simultaneously. All three instruments showed transient decreases, followed by prolonged increases in impedance. Although xCELLigence could detect these changes, it was unable to determine which component of the barrier was affected. In contrast, ECIS and cellZscope were both able to attribute responses to particular barrier components, and ECIS had a higher sensitivity than cellZscope. Finally, as cellZscope uses Transwells, it allows access to the basolateral compartment, an important advantage of this technology. Furthermore, although xCELLigence readings are equivalent to ECIS, the reduced frequency range greatly limits interpretation. This work demonstrates that instruments must be carefully selected in order to ensure that they are appropriate for the experimental questions being asked.

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Label-Free, Real-Time Measurement of Metabolism of Adherent and Suspended Single Cells by In-Cell Fourier Transform Infrared Microspectroscopy

International Journal of Molecular Sciences ◽

10.3390/ijms221910742 ◽

2021 ◽

Vol 22 (19) ◽

pp. 10742

Author(s):

Tommaso Vannocci ◽

Luca Quaroni ◽

Antonio de Riso ◽

Giulia Milordini ◽

Magda Wolna ◽

...

Keyword(s):

Real Time ◽

Mammalian Cells ◽

Single Cells ◽

Adherent Cells ◽

Label Free ◽

Ir Microscopy ◽

Infrared Microspectroscopy ◽

Real Time Measurement ◽

Fourier Transform Infrared Microspectroscopy ◽

Molecular Components

We used infrared (IR) microscopy to monitor in real-time the metabolic turnover of individual mammalian cells in morphologically different states. By relying on the intrinsic absorption of mid-IR light by molecular components, we could discriminate the metabolism of adherent cells as compared to suspended cells. We identified major biochemical differences between the two cellular states, whereby only adherent cells appeared to rely heavily on glycolytic turnover and lactic fermentation. We also report spectroscopic variations that appear as spectral oscillations in the IR domain, observed only when using synchrotron infrared radiation. We propose that this effect could be used as a reporter of the cellular conditions. Our results are instrumental in establishing IR microscopy as a label-free method for real-time metabolic studies of individual cells in different morphological states, and in more complex cellular ensembles.

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A label-free fluorescent aptasensor based on HCR and G-quadruplex DNAzymes for the detection of prostate-specific antigen

The Analyst ◽

10.1039/d0an02188a ◽

2021 ◽

Author(s):

Ruirui Zhao ◽

Lu Zhao ◽

Haidi Feng ◽

Xiaoliang Chen ◽

Huilin Zhang ◽

...

Keyword(s):

Prostate Specific Antigen ◽

Specific Antigen ◽

Label Free ◽

Fluorescence Sensing ◽

Sensing Platforms ◽

G Quadruplex ◽

Fluorescent Aptasensor

Fluorescence sensing platforms based on HCR and G-quadruplex DNAzyme amplification strategies for the detection of prostate-specific antigen.

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Label-free real-time monitoring of the BCR-triggered activation of primary human B cells modulated by the simultaneous engagement of inhibitory receptors

Biosensors and Bioelectronics ◽

10.1016/j.bios.2021.113469 ◽

2021 ◽

pp. 113469

Author(s):

Kristof Kliment ◽

Inna Szekacs ◽

Beatrix Peter ◽

Anna Erdei ◽

Istvan Kurucz ◽

...

Keyword(s):

B Cells ◽

Real Time ◽

Label Free ◽

Inhibitory Receptors ◽

Real Time Monitoring ◽

Human B Cells

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Real-time label-free detection of DNA hybridization using a functionalized graphene field effect transistor: a theoretical study

Journal of Nanoparticle Research ◽

10.1007/s11051-021-05295-1 ◽

2021 ◽

Vol 23 (8) ◽

Author(s):

Sheida Bagherzadeh-Nobari ◽

Reza Kalantarinejad

Keyword(s):

Real Time ◽

Dna Hybridization ◽

Field Effect ◽

Field Effect Transistor ◽

Theoretical Study ◽

Functionalized Graphene ◽

Label Free ◽

Graphene Field Effect Transistor ◽

Label Free Detection ◽

Effect Transistor

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Real‐Time and Label‐Free Measurement of Deubiquitinase Activity with a MspA Nanopore

ChemBioChem ◽

10.1002/cbic.202100345 ◽

2021 ◽

Author(s):

Spencer A. Shorkey ◽

Jiale Du ◽

Ryan Pham ◽

Eric R. Strieter ◽

Min Chen

Keyword(s):

Real Time ◽

Label Free

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xCELLanalyzer: A Framework for the Analysis of Cellular Impedance Measurements for Mode of Action Discovery

SLAS DISCOVERY Advancing Life Sciences ◽

10.1177/2472555218819459 ◽

2019 ◽

Vol 24 (3) ◽

pp. 213-223 ◽

Cited By ~ 1

Author(s):

Raimo Franke ◽

Bettina Hinkelmann ◽

Verena Fetz ◽

Theresia Stradal ◽

Florenz Sasse ◽

...

Keyword(s):

Data Analysis ◽

Bioactive Compounds ◽

Mode Of Action ◽

Mammalian Cells ◽

Cellular Response ◽

Label Free ◽

Synthesis Inhibitor ◽

Analysis Pipeline ◽

Bioactive Natural Products ◽

Data Analysis Pipeline

Mode of action (MoA) identification of bioactive compounds is very often a challenging and time-consuming task. We used a label-free kinetic profiling method based on an impedance readout to monitor the time-dependent cellular response profiles for the interaction of bioactive natural products and other small molecules with mammalian cells. Such approaches have been rarely used so far due to the lack of data mining tools to properly capture the characteristics of the impedance curves. We developed a data analysis pipeline for the xCELLigence Real-Time Cell Analysis detection platform to process the data, assess and score their reproducibility, and provide rank-based MoA predictions for a reference set of 60 bioactive compounds. The method can reveal additional, previously unknown targets, as exemplified by the identification of tubulin-destabilizing activities of the RNA synthesis inhibitor actinomycin D and the effects on DNA replication of vioprolide A. The data analysis pipeline is based on the statistical programming language R and is available to the scientific community through a GitHub repository.

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Modifications of the PAMONO-Sensor Help to Size and Quantify Low Number of Individual Biological and Non-Biological Nano-Particles

Engineering Proceedings ◽

10.3390/i3s2021dresden-10136 ◽

2021 ◽

Vol 6 (1) ◽

pp. 26

Author(s):

Rahat Morad Talukder ◽

Al Shahriar Hossain Rakib ◽

Julija Skolnik ◽

Zohair Usfoor ◽

Katharina Kaufmann ◽

...

Keyword(s):

Surface Plasmon Resonance ◽

Surface Plasmon ◽

Plasmon Resonance ◽

Nano Particles ◽

Label Free ◽

Specific Detection ◽

Virus Like Particles ◽

Low Concentrations ◽

Challenging Tasks ◽

Image Area

In a series of recently published works, we demonstrated that the plasmon-assisted microscopy of nano-objects (PAMONO) technique can be successfully employed for the sizing and quantification of single viruses, virus-like particles, microvesicles and charged non-biological particles. This approach enables label-free, but specific detection of biological nano-vesicles. Hence, the sensor, which was built up utilizing plasmon-assisted microscopy, possesses relative versatility and it can be used as a platform for cell-based assays. However, one of the challenging tasks for such a sensor was the ability to reach a homogeneous illumination of the whole surface of the gold sensor slide. Moreover, in order to enable the detection of even relatively low concentrations of nano-particles, the focused image area had to be expanded. Both tasks were solved via modifications of previously described PAMONO-sensor set ups. Taken together, our latest findings can help to develop a research and diagnostic platform based on the principles of the surface plasmon resonance (SPR)-assisted microscopy of nano-objects.

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