Custom AFM for X-ray beamlines:<i>in situ</i>biological investigations under physiological conditions

Gumí-Audenis, Berta; Carlà, Francesco; Vitorino, Miguel V.; Panzarella, A.; Porcar, Lionel; Boilot, M.; Guerber, Sylvain; Bernard, Pascal; Rodrigues, Mário S.; Sanz, Fausto; Giannotti, Marina I.; Costa, Luca

doi:10.1107/s1600577515016318

Cited by 15 publications

(13 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Consequently, if reasonable protein binding is observed on HOPG, it could be recommended to use hydrophobic surfaces for imaging X-ray exposed molecules, and thus detect easily the presence of radiation damage by looking at variation of protein binding on HOPG. Such apparent change in molecular surface properties has been already observed in case of lipid model membranes deposited onto silicon substrates in an in-situ X-ray - AFM combined experiment [63]. Both X-ray Reflectometry and AFM showed a deacrease of the membranes surface coverage after exposure to X-ray.…”

Section: Discussionsupporting

confidence: 58%

Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules

et al. 2016

View full text Add to dashboard Cite

BackgroundSynchrotron radiation facilities are pillars of modern structural biology. Small-Angle X-ray scattering performed at synchrotron sources is often used to characterize the shape of biological macromolecules. A major challenge with high-energy X-ray beam on such macromolecules is the perturbation of sample due to radiation damage.ResultsBy employing atomic force microscopy, another common technique to determine the shape of biological macromolecules when deposited on flat substrates, we present a protocol to evaluate and characterize consequences of radiation damage. It requires the acquisition of images of irradiated samples at the single molecule level in a timely manner while using minimal amounts of protein. The protocol has been tested on two different molecular systems: a large globular tetremeric enzyme (β-Amylase) and a rod-shape plant virus (tobacco mosaic virus). Radiation damage on the globular enzyme leads to an apparent increase in molecular sizes whereas the effect on the long virus is a breakage into smaller pieces resulting in a decrease of the average long-axis radius.ConclusionsThese results show that radiation damage can appear in different forms and strongly support the need to check the effect of radiation damage at synchrotron sources using the presented protocol.

show abstract

Section: Discussionsupporting

confidence: 58%

Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Recently, a fast AFM has been developed and successfully tested in a synchrotron beamline, extending the capabilities to biological applications [105]. In particular, simple DOPC and DPPC SLBs were first studied using the XR-AFM setup, which allowed us to evaluate radiation damage.…”

Section: Forthcoming Steps: Coupling Afm With X-ray Techniquesmentioning

confidence: 99%

“…Red : 2nd XRR; ( B ) AFM images of DPPC bilayers: ( left ) before being exposed to XR, ( right ) after being damaged by the XR beam during the acquisition of the 1st XRR (22.5 keV). Adapted with permission from [105]; ( C ) XRR curves on DPPC bilayers. Blue : 1st XRR experimental data.…”

Section: Figurementioning

confidence: 99%

Structure and Nanomechanics of Model Membranes by Atomic Force Microscopy and Spectroscopy: Insights into the Role of Cholesterol and Sphingolipids

et al. 2016

View full text Add to dashboard Cite

Biological membranes mediate several biological processes that are directly associated with their physical properties but sometimes difficult to evaluate. Supported lipid bilayers (SLBs) are model systems widely used to characterize the structure of biological membranes. Cholesterol (Chol) plays an essential role in the modulation of membrane physical properties. It directly influences the order and mechanical stability of the lipid bilayers, and it is known to laterally segregate in rafts in the outer leaflet of the membrane together with sphingolipids (SLs). Atomic force microscope (AFM) is a powerful tool as it is capable to sense and apply forces with high accuracy, with distance and force resolution at the nanoscale, and in a controlled environment. AFM-based force spectroscopy (AFM-FS) has become a crucial technique to study the nanomechanical stability of SLBs by controlling the liquid media and the temperature variations. In this contribution, we review recent AFM and AFM-FS studies on the effect of Chol on the morphology and mechanical properties of model SLBs, including complex bilayers containing SLs. We also introduce a promising combination of AFM and X-ray (XR) techniques that allows for in situ characterization of dynamic processes, providing structural, morphological, and nanomechanical information.

show abstract

“…To study biological material under physiological conditions, we have recently developed an X-AFM capable of observing the effects of radiation damage and performing AFM coupled to X-ray reflectivity on hydrated biological samples (Gumí-Audenis et al, 2015). In this work, we extend the concept of an in situ synchrotron X-AFM to the normal-incidence geometry.…”

Section: Introductionmentioning

confidence: 99%

Anin situatomic force microscope for normal-incidence nanofocus X-ray experiments

Vitorino

Fuchs

Dane

et al. 2016

J Synchrotron Radiat

Self Cite

View full text Add to dashboard Cite

A compact high-speed X-ray atomic force microscope has been developed for in situ use in normal-incidence X-ray experiments on synchrotron beamlines, allowing for simultaneous characterization of samples in direct space with nanometric lateral resolution while employing nanofocused X-ray beams. In the present work the instrument is used to observe radiation damage effects produced by an intense X-ray nanobeam on a semiconducting organic thin film. The formation of micrometric holes induced by the beam occurring on a timescale of seconds is characterized.

show abstract

Custom AFM for X-ray beamlines:in situbiological investigations under physiological conditions

Abstract: The performance of a custom atomic force microscope for grazing-incidence X-ray experiments on hydrated soft and biological samples is presented.

Cited by 15 publications

References 28 publications

Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules

Combined small angle X-ray solution scattering with atomic force microscopy for characterizing radiation damage on biological macromolecules

Structure and Nanomechanics of Model Membranes by Atomic Force Microscopy and Spectroscopy: Insights into the Role of Cholesterol and Sphingolipids

Anin situatomic force microscope for normal-incidence nanofocus X-ray experiments

Contact Info

Product

Resources

About