<i>In vivo</i>adhesion force measurements of<i>Chlamydomonas</i>on model substrates

Kreis, Christian Titus; Grangier, Alice; Bäumchen, Oliver

doi:10.1039/c8sm02236d

Cited by 32 publications

(29 citation statements)

References 59 publications

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“…A thin slab of silicon cut from a wafer (type P/Bor, orientation 100 , resistivity 120 Ωcm, unilateral polished, Si-Mat, Germany) has been glued onto a custom-made stainless steel substrate holder with PDMS. Substrate [30,33]. The micropipette cantilever, furnished with a tube and syringe filled with deionised water, was mounted onto a 3axis piezo-driven manual micromanipulator (PCS-5400, Burleigh, Thorlabs, USA) and inserted into the liquid cell.…”

Section: Methodsmentioning

confidence: 99%

“…In principle, forces can be measured simultaneously through the deflection of the micropipette. However, previous works utilizing such micropipette force sensors [30][31][32][33][34][35][36] were limited to quasistatic measurements. In this case the damping of the micropipette cantilever by hydrodynamic drag forces remains negligible, and forces can be directly recovered from the cantilever deflection and its static spring constant.…”

Section: Introductionmentioning

confidence: 99%

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Dynamic force measurements on swimming Chlamydomonas cells using micropipette force sensors

Böddeker

Karpitschka

Kreis

et al. 2020

J. R. Soc. Interface.

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Flagella and cilia are cellular appendages that inherit essential functions of microbial life including sensing and navigating the environment. In order to propel a swimming microorganism they displace the surrounding fluid by means of periodic motions, while precisely-timed modulations of their beating patterns enable the cell to steer towards or away from specific locations. Characterizing the dynamic forces, however, is challenging and typically relies on indirect experimental approaches. Here, we present direct in vivo measurements of the dynamic forces of motile Chlamydomonas reinhardtii cells in controlled environments. The experiments are based on partially aspirating a living microorganism at the tip of a micropipette force sensor and optically recording the micropipette's position fluctuations with high temporal and sub-pixel spatial resolution. Spectral signal analysis allows for isolating the cell-generated dynamic forces associated to the periodic motion of the flagella from background noise. We provide an analytic elasto-hydrodynamic model for the micropipette force sensor and describe how to obtain the micropipette's full frequency response function from a dynamic force calibration. Using this approach, we find dynamic forces during the free swimming activity of individual Chlamydomonas reinhardtii cells of 23 ± 5 pN resulting from the coordinated flagellar beating with a frequency of 51 ± 6 Hz. This dynamic micropipette force sensor (DMFS) technique generalises the applicability of micropipettes as force sensors from static to dynamic force measurements, yielding a force sensitivity in the piconewton range. In addition to measurements in bulk liquid environment, we study the dynamic forces of the biflagellated microswimmer in the vicinity of a solid/liquid interface. As we gradually decrease the distance of the swimming microbe to the interface, we measure a significantly enhanced force transduction at distances larger than the maximum extend of the beating flagella, highlighting the importance of hydrodynamic interactions for scenarios in which flagellated microorganisms encounter surfaces.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Dynamic force measurements on swimming Chlamydomonas cells using micropipette force sensors

Böddeker

Karpitschka

Kreis

et al. 2020

J. R. Soc. Interface.

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“…Here we did not observe the same behavior for active particles. This might be because i) the backflow occurs outside the Plateau borders, ii) of effects such as boundary-flagella interactions 37 that may influence the swimming efficiency of the algae or iii) of the hydrodynamics of the particle -liquid interface, which may be very different for passive particles and algae. Any (or all) of these might then compensate for the backflow effect causing a linear dependence of the velocity in the transition regime.…”

Section: Discussionmentioning

confidence: 99%

Chlamydomonas reinhardtii swimming in the Plateau borders of 2D foams

et al. 2021

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“… 10 , 11 To detach the cells efficiently and to minimize the stress on the cells, a variety of synthetic surface coatings with stimuli-responsive adhesion properties has been developed including pH responsive surfaces as well as magnetic or charge/electro-sensitive surfaces. 12 , 13 …”

Section: Introductionmentioning

confidence: 99%

Adhesion of Epithelial Cells to PNIPAm Treated Surfaces for Temperature-Controlled Cell-Sheet Harvesting

Kim

Witt

Oswald

et al. 2020

ACS Appl. Mater. Interfaces

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Stimuli responsive polymer coatings are a common motive for designing surfaces for cell biological applications. In the present study, we have characterized temperature dependent adhesive properties of poly( N -isopropylacrylamide) (PNIPAm) microgel coated surfaces (PMS) using various atomic force microscopy based approaches. We imaged and quantified the material properties of PMS upon a temperature switch using quantitative AFM imaging but also employed single-cell force spectroscopy (SCFS) before and after decreasing the temperature to assess the forces and work of initial adhesion between cells and PMS. We performed a detailed analysis of steps in the force–distance curves. Finally, we applied colloid probe atomic force microscopy (CP-AFM) to analyze the adhesive properties of two major components of the extracellular matrix to PMS under temperature control, namely collagen I and fibronectin. In combination with confocal imaging, we could show that these two ECM components differ in their detachment properties from PNIPAm microgel films upon cell harvesting, and thus gained a deeper understanding of cell-sheet maturation and harvesting process and the involved partial ECM dissolution.

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In vivoadhesion force measurements ofChlamydomonason model substrates

Abstract: A universal adhesion mechanism allows Chlamydomonas to effectively colonize abiotic surfaces, as evidenced by in vivo adhesion force measurements.

Cited by 32 publications

References 59 publications

Dynamic force measurements on swimming Chlamydomonas cells using micropipette force sensors

Dynamic force measurements on swimming Chlamydomonas cells using micropipette force sensors

Chlamydomonas reinhardtii swimming in the Plateau borders of 2D foams

Adhesion of Epithelial Cells to PNIPAm Treated Surfaces for Temperature-Controlled Cell-Sheet Harvesting

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