scholarly journals Simultaneous in vivo time-lapse stiffness mapping and fluorescence imaging of developing tissue

2018 ◽  
Author(s):  
Amelia J. Thompson ◽  
Iva K. Pillai ◽  
Ivan B. Dimov ◽  
Christine E. Holt ◽  
Kristian Franze
Keyword(s):  
Fluorescence Imaging ◽  
Temporal Dynamics ◽  
Time Lapse ◽  
Tissue Mechanics ◽  
Tissue Stiffness ◽  
Time Resolved ◽  
Force Microscopy ◽  
Atomic Force ◽  
Spatio Temporal

AbstractTissue mechanics is important for development; however, the spatio-temporal dynamics of in vivo tissue stiffness is still poorly understood. We here developed tiv-AFM, combining time-lapse in vivo atomic force microscopy with upright fluorescence imaging of embryonic tissue, to show that in the developing Xenopus brain, a stiffness gradient evolves over time because of differential cell proliferation. Subsequently, axons turn to follow this gradient, underpinning the importance of time-resolved mechanics measurements.

scholarly journals Rapid changes in tissue mechanics regulate cell behaviour in the developing embryonic brain

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Amelia J Thompson ◽  
Eva K Pillai ◽  
Ivan B Dimov ◽  
Sarah K Foster ◽  
Christine E Holt ◽  
...  
Keyword(s):  
Temporal Dynamics ◽  
Time Lapse ◽  
Time Frame ◽  
Tissue Mechanics ◽  
Tissue Stiffness ◽  
Time Resolved ◽  
Local Tissue ◽  
Close Relationship ◽  
Spatio Temporal

Tissue mechanics is important for development; however, the spatio-temporal dynamics of in vivo tissue stiffness is still poorly understood. We here developed tiv-AFM, combining time-lapse in vivo atomic force microscopy with upright fluorescence imaging of embryonic tissue, to show that during development local tissue stiffness changes significantly within tens of minutes. Within this time frame, a stiffness gradient arose in the developing Xenopus brain, and retinal ganglion cell axons turned to follow this gradient. Changes in local tissue stiffness were largely governed by cell proliferation, as perturbation of mitosis diminished both the stiffness gradient and the caudal turn of axons found in control brains. Hence, we identified a close relationship between the dynamics of tissue mechanics and developmental processes, underpinning the importance of time-resolved stiffness measurements.

scholarly journals Predicting local tissue mechanics using immunohistochemistry

2018 ◽  
Author(s):  
David E. Koser ◽  
Emad Moeendarbary ◽  
Stefanie Kuerten ◽  
Kristian Franze
Keyword(s):  
Nervous Tissue ◽  
Tissue Mechanics ◽  
Tissue Stiffness ◽  
Force Microscopy ◽  
Local Tissue ◽  
Atomic Force ◽  
Local Stiffness ◽  
Sophisticated Technology

AbstractLocal tissue stiffness provides an important signal to which cells respond in vivo. However, assessing tissue mechanics is currently challenging and requires sophisticated technology. We here developed a model quantitatively predicting nervous tissue stiffness heterogeneities at cellular resolution based on cell density, myelin and GFAP fluorescence intensities. These histological parameters were identified by a correlation analysis of atomic force microscopy-based elasticity maps of spinal cord sections and immunohistochemical stainings. Our model provides a simple tool to estimate local stiffness distributions in nervous tissue, and it can easily be expanded to other tissue types, thus paving the way for studies of the role of mechanical signals in development and pathology.

scholarly journals Dynamics of cell wall elasticity pattern shapes the cell during yeast mating morphogenesis

Open Biology ◽  
2016 ◽  
Vol 6 (9) ◽  
pp. 160136 ◽  
Author(s):  
Björn Goldenbogen ◽  
Wolfgang Giese ◽  
Marie Hemmen ◽  
Jannis Uhlendorf ◽  
Andreas Herrmann ◽  
...  
Keyword(s):  
Cell Wall ◽  
Cell Shape ◽  
Experimental Approach ◽  
Time Resolved ◽  
Force Microscopy ◽  
Cell Wall Elasticity ◽  
Atomic Force ◽  
Stiff Material ◽  
Mating Projection

The cell wall defines cell shape and maintains integrity of fungi and plants. When exposed to mating pheromone, Saccharomyces cerevisiae grows a mating projection and alters in morphology from spherical to shmoo form. Although structural and compositional alterations of the cell wall accompany shape transitions, their impact on cell wall elasticity is unknown. In a combined theoretical and experimental approach using finite-element modelling and atomic force microscopy (AFM), we investigated the influence of spatially and temporally varying material properties on mating morphogenesis. Time-resolved elasticity maps of shmooing yeast acquired with AFM in vivo revealed distinct patterns, with soft material at the emerging mating projection and stiff material at the tip. The observed cell wall softening in the protrusion region is necessary for the formation of the characteristic shmoo shape, and results in wider and longer mating projections. The approach is generally applicable to tip-growing fungi and plants cells.

scholarly journals Mechanisms of hematin crystallization and inhibition by the antimalarial drug chloroquine

2015 ◽  
Vol 112 (16) ◽  
pp. 4946-4951 ◽  
Author(s):  
Katy N. Olafson ◽  
Megan A. Ketchum ◽  
Jeffrey D. Rimer ◽  
Peter G. Vekilov
Keyword(s):  
Antimalarial Drug ◽  
Molecular Mechanisms ◽  
Physiological Concentration ◽  
Time Resolved ◽  
Force Microscopy ◽  
Atomic Structures ◽  
Future Design ◽  
Surface Sites ◽  
Atomic Force

Hematin crystallization is the primary mechanism of heme detoxification in malaria parasites and the target of the quinoline class of antimalarials. Despite numerous studies of malaria pathophysiology, fundamental questions regarding hematin growth and inhibition remain. Among them are the identity of the crystallization medium in vivo, aqueous or organic; the mechanism of crystallization, classical or nonclassical; and whether quinoline antimalarials inhibit crystallization by sequestering hematin in the solution, or by blocking surface sites crucial for growth. Here we use time-resolved in situ atomic force microscopy (AFM) and show that the lipid subphase in the parasite may be a preferred growth medium. We provide, to our knowledge, the first evidence of the molecular mechanisms of hematin crystallization and inhibition by chloroquine, a common quinoline antimalarial drug. AFM observations demonstrate that crystallization strictly follows a classical mechanism wherein new crystal layers are generated by 2D nucleation and grow by the attachment of solute molecules. We identify four classes of surface sites available for binding of potential drugs and propose respective mechanisms of drug action. Further studies reveal that chloroquine inhibits hematin crystallization by binding to molecularly flat {100} surfaces. A 2-μM concentration of chloroquine fully arrests layer generation and step advancement, which is ∼104× less than hematin’s physiological concentration. Our results suggest that adsorption at specific growth sites may be a general mode of hemozoin growth inhibition for the quinoline antimalarials. Because the atomic structures of the identified sites are known, this insight could advance the future design and/or optimization of new antimalarials.

Structural Evolution and Alignment of Cylinder-Forming PS-b-PEP Thin Films in Confinement Studied by Time-Lapse Atomic Force Microscopy

2004 ◽  
Vol 854 ◽  
Author(s):  
Qin Zheng ◽  
Dong-Chan Lee ◽  
Luping Yu ◽  
S. J. Sibener
Keyword(s):  
Thin Films ◽  
Atomic Force Microscopy ◽  
High Temperature ◽  
Intermediate State ◽  
Structural Evolution ◽  
Time Lapse ◽  
Time Resolved ◽  
Force Microscopy ◽  
Atomic Force ◽  
Sample Spot

ABSTRACTWe have utilized time-resolved high-temperature atomic force microscopy (AFM) to investigate the mechanism by which topographic templates induce alignment of cylinder-forming diblock copolymer thin films. By tracking the same sample spot during thermal annealing, we observed that the structural evolution and alignment of thin films in confinement involve an intermediate state with disordered morphology and the evolution and annihilation of disclination quadrupoles guided by the channel edges, which ultimately lead to the essentially perfect alignment of cylindrical microdomains.

Rapid, Refined, and Robust Method for Expression, Purification, and Characterization of Recombinant Human Amyloid-beta M1-42

2019 ◽  
Author(s):  
Priya Prakash ◽  
Travis Lantz ◽  
Krupal P. Jethava ◽  
Gaurav Chopra
Keyword(s):  
Amyloid Beta ◽  
Western Blots ◽  
Force Microscopy ◽  
E Coli ◽  
Atomic Force ◽  
Set Up ◽  
Short Time

Amyloid plaques found in the brains of Alzheimer’s disease (AD) patients primarily consists of amyloid beta 1-42 (Ab42). Commercially, Ab42 is synthetized using peptide synthesizers. We describe a robust methodology for expression of recombinant human Ab(M1-42) in Rosetta(DE3)pLysS and BL21(DE3)pLysS competent E. coli with refined and rapid analytical purification techniques. The peptide is isolated and purified from the transformed cells using an optimized set-up for reverse-phase HPLC protocol, using commonly available C18 columns, yielding high amounts of peptide (~15-20 mg per 1 L culture) in a short time. The recombinant Ab(M1-42) forms characteristic aggregates similar to synthetic Ab42 aggregates as verified by western blots and atomic force microscopy to warrant future biological use. Our rapid, refined, and robust technique to purify human Ab(M1-42) can be used to synthesize chemical probes for several downstream in vitro and in vivo assays to facilitate AD research.

scholarly journals Biomechanics of Neutrophil Tethers

Life ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 515
Author(s):  
Andrea Cugno ◽  
Alex Marki ◽  
Klaus Ley
Keyword(s):  
Cell Activation ◽  
Optical Trap ◽  
Biomechanical Properties ◽  
Force Microscopy ◽  
Advantages And Disadvantages ◽  
Atomic Force ◽  
Microfluidic Flow ◽  
Biomembrane Force Probe ◽  
Membrane Reservoir

Leukocytes, including neutrophils, which are propelled by blood flow, can roll on inflamed endothelium using transient bonds between selectins and their ligands, and integrins and their ligands. When such receptor–ligand bonds last long enough, the leukocyte microvilli become extended and eventually form thin, 20 m long tethers. Tether formation can be observed in blood vessels in vivo and in microfluidic flow chambers. Tethers can also be extracted using micropipette aspiration, biomembrane force probe, optical trap, or atomic force microscopy approaches. Here, we review the biomechanical properties of leukocyte tethers as gleaned from such measurements and discuss the advantages and disadvantages of each approach. We also review and discuss viscoelastic models that describe the dependence of tether formation on time, force, rate of loading, and cell activation. We close by emphasizing the need to combine experimental observations with quantitative models and computer simulations to understand how tether formation is affected by membrane tension, membrane reservoir, and interactions of the membrane with the cytoskeleton.

scholarly journals Silane-Coating Strategy for Titanium Functionalization Does Not Impair Osteogenesis In Vivo

Materials ◽  
2021 ◽  
Vol 14 (7) ◽  
pp. 1814
Author(s):  
Plinio Mendes Senna ◽  
Carlos Fernando de Almeida Barros Mourão ◽  
Rafael Coutinho Mello-Machado ◽  
Kayvon Javid ◽  
Pietro Montemezzi ◽  
...  
Keyword(s):  
Bone Formation ◽  
Photoelectron Spectroscopy ◽  
Titanium Surface ◽  
Rabbit Model ◽  
Biologically Active ◽  
Force Microscopy ◽  
Atomic Force ◽  
Silane Coating ◽  
Titanium Screws

Silane-coating strategy has been used to bind biological compounds to the titanium surface, thereby making implant devices biologically active. However, it has not been determined if the presence of the silane coating itself is biocompatible to osseointegration. The aim of the present study was to evaluate if silane-coating affects bone formation on titanium using a rabbit model. For this, titanium screw implants (3.75 by 6 mm) were hydroxylated in a solution of H2SO4/30% H2O2 for 4 h before silane-coating with 3-aminopropyltriethoxysilane (APTES). A parallel set of titanium screws underwent only the hydroxylation process to present similar acid-etched topography as a control. The presence of the silane on the surface was checked by x-ray photoelectron spectroscopy (XPS), with scanning electron microscopy (SEM) and atomic force microscopy (AFM). A total of 40 titanium screws were implanted in the tibia of ten New Zealand rabbits in order to evaluate bone-to-implant contact (BIC) after 3 weeks and 6 weeks of healing. Silane-coated surface presented higher nitrogen content in the XPS analysis, while micro- and nano-topography of the surface remained unaffected. No difference between the groups was observed after 3 and 6 weeks of healing (p > 0.05, independent t-test), although an increase in BIC occurred over time. These results indicate that silanization of a titanium surface with APTES did not impair the bone formation, indicating that this can be a reliable tool to anchor osteogenic molecules on the surface of implant devices.

scholarly journals Review of time-resolved non-contact electrostatic force microscopy techniques with applications to ionic transport measurements

2019 ◽  
Vol 10 ◽  
pp. 617-633 ◽  
Author(s):  
Aaron Mascaro ◽  
Yoichi Miyahara ◽  
Tyler Enright ◽  
Omur E Dagdeviren ◽  
Peter Grütter
Keyword(s):  
Atomic Force Microscopy ◽  
Electrostatic Force ◽  
Oscillation Period ◽  
Electrostatic Force Microscopy ◽  
Time Varying ◽  
Time Resolved ◽  
Battery Materials ◽  
Force Microscopy ◽  
Atomic Force ◽  
Microscopy Techniques

Recently, there have been a number of variations of electrostatic force microscopy (EFM) that allow for the measurement of time-varying forces arising from phenomena such as ion transport in battery materials or charge separation in photovoltaic systems. These forces reveal information about dynamic processes happening over nanometer length scales due to the nanometer-sized probe tips used in atomic force microscopy. Here, we review in detail several time-resolved EFM techniques based on non-contact atomic force microscopy, elaborating on their specific limitations and challenges. We also introduce a new experimental technique that can resolve time-varying signals well below the oscillation period of the cantilever and compare and contrast it with those previously established.

Sign in / Sign up

Export Citation Format

Share Document