901D: High-Speed Video Sarcomere Length
The High-Speed Video Sarcomere Length (HVSL) system provides high-speed, accurate sarcomere length measurement from a live video image. Our 901D HVSL system uses a high-resolution, USB 3.0 camera to provide full-frame images (1280×1024) at up to 300 frames per second. This frame rate increases to 3,000 frames per second when the camera is windowed down to 16 pixel lines.
Our software allows dynamic tracking of sarcomere length within the image which ensures that, as the tissue moves and contracts, the same sarcomeres will always be used for the calculation. Image capture and sarcomere length data can be controlled by our 600A Digital Controller software which runs on a real-time Linux operating system.
Unique Record and Playback functions allow high-speed streaming to disk and permit calculation of sarcomere length from image files. The Analysis function graphically displays sarcomere length and velocity over time.
The software includes a Scope function that provides a scrolling data display of sarcomere length with time. Researchers can accurately and easily record up to 2 hours of sarcomere length data. The Scope display allows freeze-framing at any time. In addition to scrolling through the sarcomere history, Scope also provides SL data logging to disk at rates of up to 1,000 samples per second.
The program includes two computationally accurate and fast length calculation methods, Autocorrelation Function (ACF)/Sine-fit and Fast Fourier Transform (FFT)/curve fit.
Components
901-CAM: HVSL Camera
911A: HVSL Software
901-CAL: Microscope Calibration Slide for 900B/901B
USB 3.0 Cable
Variant
901D-SA High-Speed Video Sarcomere Length Standalone System:
Standalone high-speed video sarcomere length system that includes Linux PC, high-speed camera, software and all cables
Stories of Success
900B – Sarcomere Length in Zebrafish Larvae
CHALLENGE
In 2011 Dr. Dennis Claflin approached us to try out our 900B Video Sarcomere Length system. Dr. Claflin was a long time user of Aurora Scientific equipment and an expert in single fiber measurements. He had been using a simple laser diffraction method to quantify SL in his permeabilized fibers, however, this method was not compatible with another one of his chambers.
This chamber would be mounted on an inverted microscope and used to test the mechanical forces of single, living zebrafish larvae. Because of their relative ease of rearing and their transparent body, these larvae can be used as an inexpensive model for a variety of human myopathies.
SOLUTION
Since our 900B Video Sarcomere Length system uses a C-mount camera, it is compatible with virtually all inverted microscopes. Additionally, there are no complicated procedures for calibration of different tissues. A simple reticule to correlate camera pixels to distance and 5 minute calibration was all that was required. As long as the sample presents a remotely visible sarcomere pattern the software would converge to the dominant solution. All of these attributes made the 900B the right choice to study zebrafish larvae.
RESULTS
The video sarcomere length system was put to work on a variety of different tissues such as lumbrical muscle and ultimately zebrafish. The camera proved quite successful for setting resting SL in the zebrafish larvae despite the overlapping of tissue and skin. The lab’s results using these methods have been extensively published and were recently featured in a JoVE video publication.
Select References
- Adao, Rui et al. “Urocortin-2 improves right ventricular function and attenuates pulmonary arterial hypertension.” Cardiovascular Research (2018) DOI: 10.1093/cvr/cvy076
- Tonino, Paulo et al. “The giant protein titin regulates the length of the striated muscle thick filament.” Nature communications (2017) DOI: 10.1038/s41467-017-01144-9
- Tomalka, Andre et al. “The active force-length relationship is invisible during extensive eccentric contractions in skinned skeletal muscle fibres.” Proceedings of the Royal Society B (2017) DOI: 10.1098/rspb.2016.2497
- Noonan et al. “Paraspinal Muscle Contractile Function is Impaired in the ENT1-deficient Mouse Model of Progressive Spine Pathology” Spine (2021) DOI: 10.1097/BRS.0000000000003882
- Noonan et al. “Paraspinal Muscle Contractile Function is Impaired in the ENT1-deficient Mouse Model of Progressive Spine Pathology” Spine (2021) DOI: 10.1097/BRS.0000000000003882
- Brynnel et al. “Downsizing the molecular spring of the giant protein titin reveals that skeletal muscle titin determines passive stiffness and drives longitudinal hypertrophy” eLife (2018) DOI: 10.7554/eLife.40532
- Radke, Michael H. et al. “Deleting Full Length Titin Versus the Titin M-Band Region Leads to Differential Mechanosignaling and Cardiac Phenotypes.” Circulation (2019) DOI: 10.1161/CIRCULATIONAHA.118.037588
- Land, Sander et al. “A model of cardiac contraction based on novel measurements of tension development in human cardiomyocytes.” Journal of Molecular and Cellular Cardiology (2017) DOI: 10.1016/j.yjmcc.2017.03.008
- Bagni, M. Angela et al “The effects of fatigue and oxidation on contractile function of intact muscle fibers and myofibrils isolated from the mouse diaphragm.” Scientific Reports (2019) DOI: 10.1038/s41598-019-39353-5
- Kazmierczak K. et al. “Slow-twitch skeletal muscle defects accompany cardiac dysfunction in transgenic mice with a mutation in the myosin regulatory light chain.” The FASEB Journal (2019) DOI: 10.1096/fj.201801402R
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Learn MoreSpecifications
| Software Specifications | 901D |
|---|---|
| Operating System | Linux |
| Image Functions | Brightness, Contrast, Region of Interest (ROI), Offset |
| ROI Display | Width and Height: Readout in microns or pixels Coordinate of lower left corner of ROI: pixels |
| Camera ROI Function | Sets camera Region of Interest (ROI) |
| Calibration | Field of View (FOV) or Calibration Length |
| SL Calculation Algorithm | Fast Fourier Transform/Curve Fit, Autocorrelation Function/Sine-Fit |
| Scope Function | Scrolling display of sarcomere length vs time |
| Record Function | Stores images, calculated SL values or both to disk |
| Playback Function | Playback data, calculate SL, and create AVI movie files |
| Analysis | Plots Sarcomere Length and Velocity vs Time |
| Camera Specifications | 901D |
|---|---|
| Type | CMOS Mono |
| Resolution | 1936 x 1216 (full resolution) 1920 x 1200 (default resolution) |
| Pixel Size [µm] | 3.45 x 3.45 |
| Optical Format | 1/2.3″ |
| Frame Rate | Full Frame (1920 x 1200): 164 frames per second Maximum (Reduced ROI 1920 x 8): 3,000 frames per second |
| Lens Mount | C mount |
| Interface | USB 3.0 |
| Power Required | USB Connection |
| Power Consumption [W] | 2.8 @ 5VDC |
| Weight [g] | 85 |
| Dimensions [mm] | 42.8L x 29W x 29H |