Measuring Ventral Nerve Cord Stiffness in Live Flat-dissected Drosophila Embryos by Atomic Force Microscopy

Overview

Journal STAR Protoc

Publisher Cell Press

Specialties Biology
Biomedical Engineering
Science

Date 2023 Jan 3

PMID 36595903

Authors

Katerina Karkali

Ignasi Jorba

Daniel Navajas

Enrique Martin-Blanco

Affiliations

Soon will be listed here.

Abstract

Drosophila is an amenable system for addressing the mechanics of morphogenesis. We describe a workflow for characterizing the mechanical properties of its ventral nerve cord (VNC), at different developmental stages, in live, flat-dissected embryos employing atomic force microscopy (AFM). AFM is performed with spherical probes, and stiffness (Young's modulus) is calculated by fitting force curves with Hertz's contact model. For complete details on the use and execution of this protocol, please refer to Karkali et al. (2022).

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References

Rickert C, Kunz T, Harris K, Whitington P, Technau G . Labeling of single cells in the central nervous system of Drosophila melanogaster. J Vis Exp. 2013; (73):e50150. PMC: 3622092. DOI: 10.3791/50150. View

Alcaraz J, Otero J, Jorba I, Navajas D . Bidirectional mechanobiology between cells and their local extracellular matrix probed by atomic force microscopy. Semin Cell Dev Biol. 2017; 73:71-81. DOI: 10.1016/j.semcdb.2017.07.020. View

Schillers H, Rianna C, Schape J, Luque T, Doschke H, Walte M . Standardized Nanomechanical Atomic Force Microscopy Procedure (SNAP) for Measuring Soft and Biological Samples. Sci Rep. 2017; 7(1):5117. PMC: 5505948. DOI: 10.1038/s41598-017-05383-0. View

Rico F, Roca-Cusachs P, Gavara N, Farre R, Rotger M, Navajas D . Probing mechanical properties of living cells by atomic force microscopy with blunted pyramidal cantilever tips. Phys Rev E Stat Nonlin Soft Matter Phys. 2005; 72(2 Pt 1):021914. DOI: 10.1103/PhysRevE.72.021914. View

Brent J, Werner K, McCabe B . Drosophila larval NMJ dissection. J Vis Exp. 2009; (24). PMC: 2762896. DOI: 10.3791/1107. View

Gavara N . Combined strategies for optimal detection of the contact point in AFM force-indentation curves obtained on thin samples and adherent cells. Sci Rep. 2016; 6:21267. PMC: 4759531. DOI: 10.1038/srep21267. View

Jorba I, Menal M, Torres M, Gozal D, Pinol-Ripoll G, Colell A . Ageing and chronic intermittent hypoxia mimicking sleep apnea do not modify local brain tissue stiffness in healthy mice. J Mech Behav Biomed Mater. 2017; 71:106-113. DOI: 10.1016/j.jmbbm.2017.03.001. View

Jorba I, Beltran G, Falcones B, Suki B, Farre R, Garcia-Aznar J . Nonlinear elasticity of the lung extracellular microenvironment is regulated by macroscale tissue strain. Acta Biomater. 2019; 92:265-276. PMC: 6701712. DOI: 10.1016/j.actbio.2019.05.023. View

Oliveira V, Uriarte J, Falcones B, Jorba I, Zin W, Farre R . Biomechanical Response of Lung Epithelial Cells to Iron Oxide and Titanium Dioxide Nanoparticles. Front Physiol. 2019; 10:1047. PMC: 6707084. DOI: 10.3389/fphys.2019.01047. View

10.

Crews S . Embryonic CNS Development: Neurogenesis, Gliogenesis, Cell Fate, and Differentiation. Genetics. 2019; 213(4):1111-1144. PMC: 6893389. DOI: 10.1534/genetics.119.300974. View

11.

Karkali K, Tiwari P, Singh A, Tlili S, Jorba I, Navajas D . Condensation of the Drosophila nerve cord is oscillatory and depends on coordinated mechanical interactions. Dev Cell. 2022; 57(7):867-882.e5. DOI: 10.1016/j.devcel.2022.03.007. View

12.

Andreu I, Luque T, Sancho A, Pelacho B, Iglesias-Garcia O, Melo E . Heterogeneous micromechanical properties of the extracellular matrix in healthy and infarcted hearts. Acta Biomater. 2014; 10(7):3235-42. DOI: 10.1016/j.actbio.2014.03.034. View

13.

Kuntz S, Eisen M . Drosophila embryogenesis scales uniformly across temperature in developmentally diverse species. PLoS Genet. 2014; 10(4):e1004293. PMC: 3998915. DOI: 10.1371/journal.pgen.1004293. View

14.

Sirghi L, Kylian O, Gilliland D, Ceccone G, Rossi F . Cleaning and hydrophilization of atomic force microscopy silicon probes. J Phys Chem B. 2006; 110(51):25975-81. DOI: 10.1021/jp063327g. View

15.

Hermanowicz P, Sarna M, Burda K, Gabrys H . AtomicJ: an open source software for analysis of force curves. Rev Sci Instrum. 2014; 85(6):063703. DOI: 10.1063/1.4881683. View

16.

Jorba I, Uriarte J, Campillo N, Farre R, Navajas D . Probing Micromechanical Properties of the Extracellular Matrix of Soft Tissues by Atomic Force Microscopy. J Cell Physiol. 2016; 232(1):19-26. DOI: 10.1002/jcp.25420. View

17.

Alcaraz J, Buscemi L, Grabulosa M, Trepat X, Fabry B, Farre R . Microrheology of human lung epithelial cells measured by atomic force microscopy. Biophys J. 2003; 84(3):2071-9. PMC: 1302775. DOI: 10.1016/S0006-3495(03)75014-0. View