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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.

Included Components:

  • 901-CAM: HVSL Camera
  • 911A: HVSL Software
  • 901-CAL: Microscope Calibration Slide for 900B/901B
  • USB 3.0 Cable

System Variants:

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

  • high performance at high resolution, 300 frames per second at 1280×1024 pixels
  • dynamic tracking of region of interest – keeps the same sarcomeres in the region of interest as the tissue moves and contracts
  • calculates sarcomere length accurately even with small sample regions and poor image quality
  • real-time sarcomere length measurement from digital video camera
  • easy calibration of two optical magnifications using two different methods: Field of View or Calibrated Length
  • complete package includes – HVSL Software, CMOS camera, all required cables, calibration slide.  Install software on 600A PC or purchase standalone PC
  • Record and Playback functions that allow streaming to disk at full camera speed (up to 3,000 frames per second) and calculation of sarcomere length from image files
  • Scope function provides real-time scrolling data display, stores up to 2 hours of sarcomere length data, can also log data to disk at up to 1,000 samples per second
  • SL control, image capture and SL data output can all be controlled by 600A Digital Controller
  • two computationally accurate and extremely fast length calculation methods: Autocorrelation Function (ACF)/Sine-fit and Fast Fourier Transform (FFT)/curve fit

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.

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