1600A

Permeabilized Myocyte System – Microscope Mountable

The 1600A Permeabilized Myocyte Test System is designed to simplify the complex testing of contractile properties of skinned myocytes. A large cell mounting area provides ample room for cell attachment. Push button controlled motorized stages simplifies cell attachment to the high-speed servo motor (315C, 322C) and precision force transducer (400 series).

A temperature controlled 8-well bath plate allows for seamless transition of the myocyte between varying calcium concentrations. Direct mounting on an inverted microscope stage allows for observation and sarcomere spacing detection. An optional prism reticule can be included to provide accurate measurement of cell cross section.

With the 1600A, force, length and sarcomere spacing (with optional HVSL/VSL) can all be controlled and measured easily and repeatedly. This permits measurement of simple force-pCa to more complex mechanical characterization of the cell including kTr, length-tension, force-velocity and stiffness.

The system is truly turnkey, including components for data acquisition and analysis as well. Our Digital controller and Software Analysis package (600A) comes complete with a large library of standardized protocols to make the above characterizations straightforward. Choose the 1600A system for performance, precision and progress.

Inverted microscope with 803B Permeabilized Myocyte Apparatus

Inverted microscope with Permeabilized Myocyte Apparatus

Eight temperature controlled bath wells

803B direct mounting on inverted microscope

Direct mounting of apparatus on inverted microscope

complete test system for permeabilized myocytes
temperature controlled 8-well bath plate (sizes from 400µL to 1900µL)
large cell mounting area and motorized XYZ micro-positioning stages with computerized control
control and measure force, length and sarcomere spacing (with optional 900B/901C)

can be integrated with Aurora Scientific 900B/901C Sarcomere Length Software (VSL/HVSL)
simple mounting to an inverted microscope
measure force-pCA, kTR, stiffness, length-tension and force-velocity relationships
optional optical prism included for measurement of cell depth

Stories of Success

1600A – Turnkey Solution Aids Characterization of Cardiac Cell Mechanics in Heart Failure

CHALLENGE

Dr. Jonathan Kirk was working as a postdoctoral fellow in Dr. David Kass’ lab at Johns Hopkins University who had acquired his own custom built equipment for testing mechanics of heart failure in skinned cardiomyocytes. When Dr. Kirk secured a faculty position at Loyola University in Chicago, he did not have the means to fabricate his own custom apparatus and stages to characterize cardiomyocyte function. He also needed to find a way to integrate his own stepper motor for small-scale length changes in single cell preparations while measuring and monitoring sarcomere length.

SOLUTION

Aurora Scientific had worked closely with Dr. Kirk and Dr. Kass previously so we knew we would be able to provide him a complete solution for his new lab. The 1600A Permeabilized Myocyte System was the answer as it included our 803B 8-well skinned myocyte apparatus for various calcium concentrations, 3-axis motorized stages and a complete data acquisition and analysis system. Furthermore, Aurora Scientific provided our 400A series high-resolution force transducer and incorporated our video sarcomere length software (VSL) and camera for measuring and monitoring sarcomere spacing.

RESULTS

The permeabilized myocyte system was provided to Dr. Kirk, which was easily mounted on an inverted microscope and combined with motorized stages to deliver precise control of the transducer and motor for cell attachment and testing. Furthermore, the 8-well bath plate allowed for quick determination of force-pCA in these cells. Additionally, our data acquisition software and VSL enabled full integration of Dr. Kirk’s stepper motor for precise length changes, easily monitored using the VSL camera. The 1600A has provided Dr. Kirk with a system that will allow him to characterize mechanics of heart failure and discover the mechanisms involved, leading the way to the development of potentially ground-breaking treatments for humans with this condition.

Select References

Alegre-Cebollada, Jorge et al. “S-glutathionylation of cryptic cysteines enhances titin elasticity by blocking protein folding.” Cell (2014) DOI: 10.1016/j.cell.2014.01.056
Krysiak et al. “Protein phosphatase 5 regulates titin phosphorylation and function at a sarcomere-associated mechanosensor complex in cardiomyocytes” Nature communications (2018) DOI: 10.1038/s41467-017-02483-3
Gonçalves-Rodrigues et al. “In Vitro Assessment of Cardiac Function Using Skinned Cardiomyocytes” Journal of Visualized Experiments (2020) DOI: 10.3791/60427
Pappritz et al. “Impact of syndecan-2-selected mesenchymal stromal cells on the early onset of diabetic cardiomyopathy in diabetic db/db mice” Frontiers in Cardiovascular Medicine (2021) DOI: 10.3389/fcvm.2021.632728
Pappritz et al. “Impact of syndecan-2-selected mesenchymal stromal cells on the early onset of diabetic cardiomyopathy in diabetic db/db mice” Frontiers in Cardiovascular Medicine (2021) DOI: 10.3389/fcvm.2021.632728
Kötter, Sebastian et al. “Human myocytes are protected from titin aggregation-induced stiffening by small heat shock proteins.” The Journal of Cell Biology (2014) DOI: 10.1083/jcb.201306077
Horiguchi, Hiroshi et al. “Fabrication and evaluation of reconstructed cardiac tissue and its application to bio-actuated microdevices.” IEEE Transactions on NanoBioscience (2009) DOI: 10.1109/TNB.2009.2035282
Kolijn et al. “Enhanced Cardiomyocyte Function in Hypertensive Rats With Diastolic Dysfunction and Human Heart Failure Patients After Acute Treatment With Soluble Guanylyl Cyclase (sGC) Activator” Frontiers in Physiology (2020) DOI: 10.3389/fphys.2020.00345
Chung, Eunhee and Gary M. Diffee. “Effect of aging on power output properties in rat skinned cardiac myocytes.” The Journals of Gerontology Series A: Biological Sciences and Medical Sciences (2011) DOI: 10.1093/gerona/glr150
Waddingham et al. “Diastolic dysfunction is initiated by cardiomyocyte impairment ahead of endothelial dysfunction due to increased oxidative stress and inflammation in an experimental prediabetes model” Journal of Molecular and Cellular Cardiology (2019) DOI: 10.1016/j.yjmcc.2019.10.005
Hamdani, Nazha et al. “Deranged myofilament phosphorylation and function in experimental heart failure with preserved ejection fraction.” Cardiovascular Research (2013) DOI: 10.1093/cvr/cvs353
Rivas-Pardo et al. “A HaloTag-TEV genetic cassette for mechanical phenotyping of proteins from tissues” Nature communications (2020) DOI: 10.1038/s41467-020-15465-9

System Components

803B: Permeabilized Myocyte Apparatus – Microscope Mountable

608C: LCD Monitor

A 22” widescreen LCD monitor providing high quality visualization of data collection

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Resources

Parts & Accessories

Model #	1600A
900B	Video Sarcomere Length (VSL) measurement system includes camera and VSL software that integrates with the 600A Digital Controller. The included USB camera operates at a frame rate of 80 frames per second (FPS) for full frames and up to a maximum of 1000 FPS for reduced frame sizes. The camera mounts via a C-mount adapter (not provided).
901B	High-Speed Video Sarcomere Length (HVSL) measurement system includes camera and HVSL software that integrates with the 600A Digital Controller. The included Gigabit Ethernet camera operates at a frame rate of 300 frames per second (FPS) for full frames and up to a maximum of 4700 FPS for reduced frame sizes. The camera mounts via a C-mount adapter (not provided).
830A	Half-rack adapter that allows half-rack instruments to be mounted in standard 19” racking. Works with models: 200B, 315C/322C, 400A series, 701C and 825A.
831A	Heavy gauge, steel desktop or shelf-top rack, with standard 19” width for any Aurora Scientific Instruments. Available in various heights. (6U high for 1200A and 1300A, 5U high for 1400A, 1500A and 1600A systems).