Combined Atomic Force Microscope and Volumetric Light Sheet System for Mechanobiology
Combined Atomic Force Microscope and Volumetric Light Sheet System for Mechanobiology
ABSTRACT The central goals of mechanobiology are to understand how cells generate force and how they respond to environmental mechanical stimuli. A full picture of these processes requires high-resolution, volumetric imaging with time-correlated force measurements. Here we present an instrument that combines an open-top, single-objective light sheet fluorescence microscope with an atomic force microscope (AFM), providing simultaneous volumetric imaging with high spatiotemporal resolution and high dynamic range force capability (10 pN – 100 nN). With this system we have captured lysosome trafficking, vimentin nuclear caging, and actin dynamics on the order of one second per volume. To showcase the unique advantages of combining Line Bessel light sheet imaging with AFM, we measured the forces exerted by a macrophage during Fc?R-mediated phagocytosis while performing both sequential two-color, fixed plane and volumetric imaging of F-actin. This unique instrument allows for a myriad of novel studies investigating the coupling of cellular dynamics and mechanical forces.
Falvo M.R.、Boyce M.、Superfine R.、Hobson C.M.、Hsiao J.P.、O?ˉBrien E.T. III、Watanabe T.、Grinstein S.、Nelsen E.、Hahn K.M.、Kern M.E.、Condon B.M.
Dept. of Physics and Astronomy, University of North Carolina at Chapel HillDept. of Biochemistry, Duke University School of MedicineDept. of Applied and Materials Science, University of North Carolina at Chapel HillDept. of Physics and Astronomy, University of North Carolina at Chapel HillDept. of Physics and Astronomy, University of North Carolina at Chapel HillDept. of Physics and Astronomy, University of North Carolina at Chapel HillDept. of Pharmacology, University of North Carolina at Chapel HillProgram in Cell Biology, Hospital for Sick ChildrenDept. of Physics and Astronomy, University of North Carolina at Chapel HillDept. of Pharmacology, University of North Carolina at Chapel HillDept. of Physics and Astronomy, University of North Carolina at Chapel HillDept. of Biochemistry, Duke University School of Medicine
生物科学研究方法、生物科学研究技术细胞生物学生物物理学
Falvo M.R.,Boyce M.,Superfine R.,Hobson C.M.,Hsiao J.P.,O?ˉBrien E.T. III,Watanabe T.,Grinstein S.,Nelsen E.,Hahn K.M.,Kern M.E.,Condon B.M..Combined Atomic Force Microscope and Volumetric Light Sheet System for Mechanobiology[EB/OL].(2025-03-28)[2025-05-11].https://www.biorxiv.org/content/10.1101/812396.点此复制
评论