Mfp 3d bio software

Adult female flies were anesthetized and immobilized on 25-mm glass coverslips with a thin layer of vacuum grease ventral-side-up. The heart tube was exposed via microsurgery as previously described (Ocorr et al., 2007 (link)) with additional micropipette aspiration to remove all ventral tissue proximal to the conical chamber. Each coverslip is mounted on a Fluid Cell Lite coverslip holder (Asylum Research, Goleta, CA, USA) with 1 mL of hemolymph. Hearts are checked for regular contractions to ensure they are in good health and then resubmerged in 10 mm ethylene glycol tetraacetic acid (EGTA)-treated hemolymph to arrest contraction. Prior to indentation, probes were calibrated via thermal noise method in MFP-3D Bio software (Asylum Research). Nanoindentation was performed on an MFP-3D Bio Atomic Force Microscope (Asylum Research) mounted on a Ti-U fluorescent inverted microscope (Nikon Instruments, Melville, NY, USA) with 120 pN nm⁻¹ silicon nitride cantilevers with premounted, 2-μm radius borosilicate glass spheres (Novascan Technologies, Ames, IA, USA). All indentation curves were analyzed to calculate myocardial stiffness using previously published, automated software custom-written in MATLAB (MathWorks, Natick, MA, USA; Kaushik et al., 2011 (link), 2012 (link)).