Dynamic Force Spectroscopy Analysis on the Redox States of Protein Disulphide Bonds

Ju, Lining Arnold

doi:10.1007/978-1-4939-9187-7_7

Cited by 12 publications

(14 citation statements)

References 29 publications

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“…2 a, top). After RBCs were injected into the chamber, a fashioned borosilicate micropipette was anchored on the micropipette holder and inserted into the chamber by tuning the micro-manipulator as previously described (Chen et al 2015 ; Ju 2019 ). The micropipette was then approached to grab the target RBC (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Tips of micropipettes were cut and fashioned to 1 µm diameters using a MicroForge (Model MF-900; Narishige International, USA) equipped with a 20 × eyepiece. For detailed operation procedures, please refer to the previously published protocols (Ju 2019 ).…”

Section: Methodsmentioning

confidence: 99%

“…This step can ensure the absence of air bubble inside the tube connecting the micropipette holder and the water reservoir. Then a fully filled borosilicate micropipette fabricated from the previous section was inserted into the holder tip and the holder screw was tightened (Ju 2019 ). Finally, due to the tightness of the micropipette–reservoir connection, the aspiration pressure of the borosilicate micropipette was finely controlled by adjusting the height of the connected water reservoir related to the tip of the micropipette in a stepwise manner.…”

Section: Methodsmentioning

confidence: 99%

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Fluorescence-coupled micropipette aspiration assay to examine calcium mobilization caused by red blood cell mechanosensing

et al. 2022

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Mechanical stimuli such as tension, compression, and shear stress play critical roles in the physiological functions of red blood cells (RBCs) and their homeostasis, ATP release, and rheological properties. Intracellular calcium (Ca2+) mobilization reflects RBC mechanosensing as they transverse the complex vasculature. Emerging studies have demonstrated the presence of mechanosensitive Ca2+ permeable ion channels and their function has been implicated in the regulation of RBC volume and deformability. However, how these mechanoreceptors trigger Ca2+ influx and subsequent cellular responses are still unclear. Here, we introduce a fluorescence-coupled micropipette aspiration assay to examine RBC mechanosensing at the single-cell level. To achieve a wide range of cell aspirations, we implemented and compared two negative pressure adjusting apparatuses: a homemade water manometer (− 2.94 to 0 mmH2O) and a pneumatic high-speed pressure clamp (− 25 to 0 mmHg). To visualize Ca2+ influx, RBCs were pre-loaded with an intensiometric probe Cal-520 AM, then imaged under a confocal microscope with concurrent bright-field and fluorescent imaging at acquisition rates of 10 frames per second. Remarkably, we observed the related changes in intracellular Ca2+ levels immediately after aspirating individual RBCs in a pressure-dependent manner. The RBC aspirated by the water manometer only displayed 1.1-fold increase in fluorescence intensity, whereas the RBC aspirated by the pneumatic clamp showed up to threefold increase. These results demonstrated the water manometer as a gentle tool for cell manipulation with minimal pre-activation, while the high-speed pneumatic clamp as a much stronger pressure actuator to examine cell mechanosensing directly. Together, this multimodal platform enables us to precisely control aspiration and membrane tension, and subsequently correlate this with intracellular calcium concentration dynamics in a robust and reproducible manner.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Fluorescence-coupled micropipette aspiration assay to examine calcium mobilization caused by red blood cell mechanosensing

et al. 2022

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“…Towards a further understanding of the structure–function relationship with respect to the N-AIM of VWF-A1, we employed the DFS technique—BFP to characterize the VWF-A1–GPIbα interaction in a bead–bead mode as previously established. 58 VWF-A1 was coated on a glass bead (probe) attached to the apex of a micropipette-aspirated red blood cell (RBC) ( Fig. 2a and b , left), and GPIbα was immobilized on another glass bead (Target) aspirated using an apposing micropipette ( Fig.…”

Section: Resultsmentioning

confidence: 99%

The N-terminal autoinhibitory module of the A1 domain in von Willebrand factor stabilizes the mechanosensor catch bond

Zhao

Wang

et al. 2022

RSC Chem. Biol.

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“…Figure 1D demonstrates various applications of bio-AFM (Hoh and Hansma 1992;Shao and Yang 1995;Czajkowsky et al, 2000;Hansma 2001;Goldsbury and Scheuring 2002;Malkin et al, 2002;Alonso and Goldmann 2003;Besch et al, 2003;Gadegaard 2006;Shahin and Barrera 2008;Goldsbury et al, 2009;Ramachandran et al, 2011;Kreplak 2016;Dufrene et al, 2017;Braet and Taatjes 2018;Gao et al, 2018;Cheong et al, 2019;Nandi and Ainavarapu 2021), and we are about to summarize these in this review. Unlike the AFM imaging modes where the probe is scanned over the surface of the substrate, the cantilever tip is first approached toward the substrate until tip-sample contact happens and then retracted in AFM dynamic force spectroscopy (AFM-DFS) (Sulchek et al, 2005;Neuert et al, 2006;Thormann et al, 2006;Diezemann and Janshoff 2008;Alessandrini et al, 2012;Sengupta et al, 2014;Sluysmans et al, 2018;Ju 2019;Reiter-Scherer et al, 2019;Alhalhooly et al, 2021) and single-molecule force spectroscopy (AFM-SMFS) experiments. These two are similar methods with slight differences.…”

Section: Introductionmentioning

confidence: 99%

Biosensing, Characterization of Biosensors, and Improved Drug Delivery Approaches Using Atomic Force Microscopy: A Review

Sarkar

2022

Front. Nanotechnol.

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Since its invention, atomic force microscopy (AFM) has come forth as a powerful member of the “scanning probe microscopy” (SPM) family and an unparallel platform for high-resolution imaging and characterization for inorganic and organic samples, especially biomolecules, biosensors, proteins, DNA, and live cells. AFM characterizes any sample by measuring interaction force between the AFM cantilever tip (the probe) and the sample surface, and it is advantageous over other SPM and electron micron microscopy techniques as it can visualize and characterize samples in liquid, ambient air, and vacuum. Therefore, it permits visualization of three-dimensional surface profiles of biological specimens in the near-physiological environment without sacrificing their native structures and functions and without using laborious sample preparation protocols such as freeze-drying, staining, metal coating, staining, or labeling. Biosensors are devices comprising a biological or biologically extracted material (assimilated in a physicochemical transducer) that are utilized to yield electronic signal proportional to the specific analyte concentration. These devices utilize particular biochemical reactions moderated by isolated tissues, enzymes, organelles, and immune system for detecting chemical compounds via thermal, optical, or electrical signals. Other than performing high-resolution imaging and nanomechanical characterization (e.g., determining Young’s modulus, adhesion, and deformation) of biosensors, AFM cantilever (with a ligand functionalized tip) can be transformed into a biosensor (microcantilever-based biosensors) to probe interactions with a particular receptors of choice on live cells at a single-molecule level (using AFM-based single-molecule force spectroscopy techniques) and determine interaction forces and binding kinetics of ligand receptor interactions. Targeted drug delivery systems or vehicles composed of nanoparticles are crucial in novel therapeutics. These systems leverage the idea of targeted delivery of the drug to the desired locations to reduce side effects. AFM is becoming an extremely useful tool in figuring out the topographical and nanomechanical properties of these nanoparticles and other drug delivery carriers. AFM also helps determine binding probabilities and interaction forces of these drug delivery carriers with the targeted receptors and choose the better agent for drug delivery vehicle by introducing competitive binding. In this review, we summarize contributions made by us and other researchers so far that showcase AFM as biosensors, to characterize other sensors, to improve drug delivery approaches, and to discuss future possibilities.

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Dynamic Force Spectroscopy Analysis on the Redox States of Protein Disulphide Bonds

Cited by 12 publications

References 29 publications

Fluorescence-coupled micropipette aspiration assay to examine calcium mobilization caused by red blood cell mechanosensing

Fluorescence-coupled micropipette aspiration assay to examine calcium mobilization caused by red blood cell mechanosensing

The N-terminal autoinhibitory module of the A1 domain in von Willebrand factor stabilizes the mechanosensor catch bond

Biosensing, Characterization of Biosensors, and Improved Drug Delivery Approaches Using Atomic Force Microscopy: A Review

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