RESUMO
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 single-cell volume. To showcase the unique advantages of combining Line Bessel light sheet imaging with AFM, we measured the forces exerted by a macrophage during FcɣR-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.
Assuntos
Macrófagos/química , Microscopia de Força Atômica/métodos , Microscopia de Fluorescência/métodos , Actinas/química , Actinas/metabolismo , Animais , Fenômenos Biomecânicos , Fluorescência , Células HeLa , Humanos , Macrófagos/citologia , Macrófagos/metabolismo , Camundongos , Fagocitose , Células RAW 264.7RESUMO
We investigated a case of maxillary sinus infection caused by the medusoid mutant of the basidiomycetous fungus Schizophyllum commune. Morphologies of both typical and medusoid forms of S. commune were determined. Prior reports of pathogenic homobasidiomycetes are discussed.