Mechanical properties of cardiomyocytes were assessed using an IonOptix™ SoftEdge MyoCam® system (IonOptix Corporation, Milton, MA, USA). Cardiomyocytes were placed in a chamber mounted on the stage of an Olympus IX-70 microscope and superfused (~2 ml/min at 25 °C) with a KHB buffer containing 1 mM CaCl2. Myocytes were field stimulated at 0.5 Hz. Cell shortening and relengthening were assessed including peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR90) and maximal velocities of shortening/relengthening (± dL/dt) (22 (link)).
Softedge myocam system
Manufactured by IonOptix
Sourced in United States
The SoftEdge MyoCam system is a lab equipment product that provides high-resolution imaging and analysis capabilities for the study of muscle tissue mechanics. The system is designed to capture and analyze the contractile properties of muscle samples with precision and accuracy.
Automatically generated - may contain errors
Lab products found in correlation
Softedge software, by IonOptix (6 mentions)
Myocam camera, by IonOptix (6 mentions)
Ix70 microscope, by Olympus (3 mentions)
Dual excitation fluorescence photomultiplier system, by IonOptix (3 mentions)
Liberase blendzyme 4, by Roche (2 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Liberase dh, by Roche (1 mentions)
Fura 2 am, by Dojindo Laboratories (1 mentions)
Ix70 inverted microscope, by Olympus (1 mentions)
Ix 70, by Olympus (1 mentions)
Fluo 2am, by Thermo Fisher Scientific (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Sequoia c512, by Siemens (1 mentions)
Liberase blendzyme, by Roche (1 mentions)
Ls004188, by Worthington (1 mentions)
Ls004176, by Worthington (1 mentions)
Caspase assay kit, by Beyotime (1 mentions)
32 protocols using softedge myocam system
1
Mechanical Properties of Cardiomyocytes
2
Mechanical Properties of Cardiomyocytes
3
Sarcomere Contractility and Calcium Dynamics
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Our experiment used video-based sarcomere contractility and calcium recording module in a SoftEdge MyoCam System (IonOptix Corporation, Milton, MA, USA) to assess sarcomere shortening and Ca2+ transients, as previously reported [11 (link)]. Firstly, isolated ARVMs were incubated with Fura-2/AM (2 μM) for 15 min, then the cells were washed twice with solution A. Subsequently, the ARVMs were placed in a Warner chamber mounted on the stage of an inverted microscope (Olympus, IX-70) and perfused with different solutions at a rate of 1.5 mL/min. The experiment was grouped as follows: (a) control group: solution A for 30 min; (b) Sal A group: solution A with 1 μM Sal A for 10 min; (c) ATO group: solution A with 100 μM ATO for 20 min; and (d) ATO+Sal A group: solution A with 100 μM ATO for 20 min after solution A with 1 μM Sal A for 10 min. The ARVMs were allowed to stabilize for approximately 10 min before being analyzed. In our studies, 30–40 cells from 5 rats per group were obtained. The recording and analysis of data were conducted using IonWizard software (version 6.2.0.59, Milton, MA, USA).
4
Cardiomyocyte Contractile Capacity Evaluation
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Cardiomyocyte contractile capacity was evaluated with the assistance of a SoftEdge MyoCam system (IonOptix Corporation, Milton, MA, USA) coupled with an IX-70 Olympus microscope [32 ]. In brief, cells were paced at 0.5 Hz before mechanical properties were recorded including maximal velocities of shortening/relengthening (±dL/dt), peak shortening (PS), time-to-peak shortening (TPS), and time-to-90% relengthening (TR90).
5
Echocardiography and Cardiomyocyte Analyses in Mice
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Echocardiography was performed on all mice using an Esaote Twice System with an SL3116 transducer [38 (link)]. Two-dimensional and M-mode echocardiographic measurements were recorded with a VEVO 2100 high-resolution in vivo imaging system [39 (link)]. Cardiomyocyte shortening and relengthening assays were performed using a SoftEdge MyoCam system (IonOptix, Milton, MA), as described in a previous study [40 (link)]. Briefly, cardiomyocytes were isolated from reperfused hearts, and then single cardiomyocytes were observed under an inverted microscope. The time to peak shortening (TPS), time to 90% relengthening (TR90), maximal velocity of shortening (+dL/dt), and maximal velocity of relengthening (-dL/dt) were recorded [41 (link)].
6
Isolation and Characterization of Adult Mouse Cardiomyocytes
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After echocardiographic evaluation, the ventricles were immediately removed from the hearts of WT, HE, and HO mice under anesthesia and were digested with collagenase buffer. Adult mouse cardiomyocyte isolation was performed by the described methods (Ackers‐Johnson et al., 2016 (link); Jiang et al., 2020 (link)). The only cardiomyocytes chosen for usage were those with rod shapes and distinct edges. An inverted microscope (IX‐70, Olympus) was utilized to visualize cardiomyocytes, and a SoftEdge MyoCam system (IonOptix Corporation) was utilized to assess their mechanical characteristics. The following indicators were utilized to quantify cardiomyocyte contraction and relengthening profiles: resting cell length; peak shortening (PS, normalized to the resting cell length); time‐to‐PS (TPS); maximal rates of shortening (+dL/dt) and relengthening (−dL/dt) and time‐to‐90% relengthening (TR90).
7
Isolation and Functional Analysis of Cardiomyocytes
8
Measuring Intracellular Calcium Dynamics
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NRCMs were plated on coverslips in 3-cm dishes for routine cultivation. After carrying out all intervention protocols, NRCMs were eluted with Hanks Balanced Salt Solution (HBSS), and then incubated with 5 μM/mL Fura 2-AM (Dojindo Laboratories, Kumamoto Japan) in DMSO and diluted in HBSS for 30 min at 37 °C followed by another HBSS elution. NRCMs on coverslips were then sent for calcium transient recording using a dual-excitation fluorescence photomultiplier system (IonOptix, Milton, MA, USA). Briefly, the NRCMs were continuously covered with warm oxygenated Tyrode's solution (140 mM NaCl, 1 mM MgCl2, 6 mM KCl, 10 mM glucose, 2 mM CaCl2, and 5 mM HEPES, pH 7.4) [35 (link)]. Fluorescence intensity at 510 nm was detected with alternate scanning by fluorescence emissions at 340 nm and 380 nm. Intracellular Ca2+ levels and the intracellular Ca2+ transient decay time constant were recorded and analyzed by the SoftEdge MyoCam® system (IonOptix).
9
Cardiomyocyte Mechanical Properties Assessment
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The mechanical properties of myocytes were assessed using a SoftEdge Myocam system (IonOptix, Milton, MA). SoftEdge software was used to capture changes in cardiomyocyte length during shortening and re-lengthening. The myocytes were photographed with a MyoCam camera and displayed on a computer monitor. Cell shortening and re-lengthening were assessed using the following indices: peak shortening (PS), the amplitude myocytes shortened on electrical stimulation, which is indicative of peak ventricular contractility; time-to-PS (TPS), the duration of myocyte shortening, which indicates duration of contraction; time-to-90% relengthening (TR90), which represents the duration of cardiomyocyte relaxation (90% was used rather than 100% to avoid a noisy signal at baseline concentration), and maximal velocity of shortening (+ dL/dt) and relengthening (− dL/dt), the maximal slope (derivative) of the shortening and relengthening phases, which indicate the maximal velocities of rise and fall of ventricular pressure.
10
Assessment of Cardiomyocyte Mechanics and Calcium Signaling
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Mechanical properties of cardiomyocytes were assessed using a Softedge MyoCam system (IonOptix Corporation, Milton, MA, USA) equipped with an IX-70 Olympus inverted microscope. Cardiomyocytes were electrically stimulated at 0.5 Hz in a contractile buffer containing NaCl 135 mM, KCl 4.0 mM, CaCl2 1.0 mM, MgCl 1.0 mM, glucose 10 mM and HEPES 10 mM. Cell shortening was assessed including peak shortening (PS), maximal velocity of shortening (+dL/dt), maximal velocity of re-lengthening (−dL/dt), time-to-PS (TPS), and time-to-90% re-lengthening (TR90). For intracellular Ca2+ recording, cardiomyocytes were loaded with Fura-2/AM (0.5 μM) for 10 min, and fluorescence measurements were recorded with a dual-excitation fluorescence photomultiplier system (IonOptix). To assess intracellular Ca2+ signaling, cells were exposed to light emitted by a 75-W lamp and passed through 360 nm or a 380 nm filter, while being stimulated to contract at 0.5 Hz. Fluorescence emissions were detected between 480 and 520 nm and the alterations in fura-2 fluorescence intensity (FFI) were quantitated from the FFI ratio at 360 nm to 380 nm. Fluorescence decay time was assessed as an indicator of intracellular Ca2+ clearing [57 (link), 61 (link)].
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