950A: Calcium Fluorescence System

The 950A calcium fluorescence system enables physiology researchers to easily measure intracellular calcium along with tissue length, force and sarcomere length. Powerful twitch-based software controls all instruments and collects and analyzes the data. The entire system can be automated to perform entire experiments in a repeatable fashion.

The 955A FluoroTrack software controls the IonOptix fluorescence photometry equipment and collects the fluorescence signals from the photomultiplier. Test protocols written using the 600A Protocol Editor control the entire experiment including the stimulator, high-speed length controller, force transducer, apparatus, fluorescence equipment and sarcomere length measurement. Data collection from all of these instruments is stored in a single time-stamped file. This highly integrated system allows a single computer to control and store data from many different instruments including those from Aurora Scientific, IonOptix and Sutter (DG-4 light source).

Powerful Twitch Analysis provides an easy-to-use analysis and data export capability. Length, Force, SL, Ratio and Calcium data streams can be analyzed to provide average twitch data and statistics.

  • integrates with various Aurora Scientific systems to provide simultaneous acquisition of fluorescence photometry with length, force, and sarcomere length

  • controls all Aurora Scientific instruments plus light sources from major manufacturers (IonOptix HyperSwitch and Sutter Lambda DG-4)

  • complete fluorescence photometry system includes hardware and software

  • twitch based data acquisition and analysis ideal for cardiac studies, includes stimulator control

  • system can be configured to work with any indicator fluorophore (BCECF, Fluo-3/4, Fura-2, Indo-1)

  • two operating modes for Fura-2 – interleaved and interpolated numerator

  • calculate ratio or calcium (Ca2+) concentration at up to 1000 samples per second

Components

955A: FluoroTrack – Calcium Fluorescence Software
960A: FluoroDAQ Data Acquisition Controller
961A: HyperSwitch Dual Excitation Light Source
962A: Cell Framing Adapter

962B: CFA Option D: Dual Emission
963A: Photomultiplier Subsystem
964-L: Microscope Components – Leica
964-N: Microscope Components – Nikon

964-O: Microscope Components – Olympus
964-Z: Microscope Components – Zeiss
965A Option S: 75W Xenon Light Source

Stories of Success

950A – Software Module Helps Synchronize Cutting Edge Data In Myocyte Research

CHALLENGE

In late 2013 Dr. Chris Ward of the University of Maryland was working on improving experimental techniques for measuring force, SL and calcium release in single, living myocytes from the FDB muscle.  Dr. Ward had been a long time user of our equipment and was already using our 600A and HVSL software programs to record force and SL.  However, he had been using another lab built solution to measure calcium and was searching for a way to easily synchronize all three measurements.

SOLUTION

The Aurora Scientific FluoroTrack software proved to be the missing piece.  FluoroTrack allowed real-time monitoring of calcium transients and integrated seamlessly with the existing HVSL and 600A programs. In addition, Aurora Scientific made minor customizations to be compatible with Dr. Ward’s existing light source and calcium measurement instruments.

RESULTS

After making these configurations Dr. Ward was able to begin collecting synchronized data almost immediately. This integration provided a single, time-stamped file for easy collection and analysis of force, SL and calcium from FDB cells. The data collected were presented at a Biophysical society meeting some short 2 months later and has gone on to be published.  The system is still an integral part of Dr. Ward’s research and the software continues to evolve and be improved thanks to feedback from Dr. Ward’s lab and other researchers.

Select References

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Specifications

Software Specifications 950A
Operating System Linux – RTAI, real-time
Companion Software 600A, 602A, 802D, 820A, 900B, 901C
Shutter Control Remote control of shutter
Stimulation Control Manual or Automatic stimulation control – frequency (Hz. or BPM), pulse
width, pulse duration
Data Channels Recorded Numerator, Denominator, Ratio, Ca2+, Length, Force, SL, Stimulation Timing
Background Light Measurement Simple, easy-to-use measurement of background light level
Scrolling Data Display Real-time data output of chosen channels. Displays up to 4 hours of data –
zoom function, time base control, pause function
Data Tags Create data tags with mouse or F-key, create tags during data collection or
after, place tags at chosen time
Twitch Analysis Displays Time Series, Twitch Based, Data Records in table view, Tags in table view,
Tags shown on Time Series and Twitch Based views
Twitch Analysis Easily choose twitches of interest
Twitch Analysis – calculates average twitch parameters, calculates twitch
statistics: baseline, peak, peak ht, peak %, peak ht%, max dX/dt, min dX/dt, T
dX/dt max, T dX/dt min, T peak, T 10% rise, T 50% rise, T 90% rise, dT 10-
90%, T 90% fall, T 50% fall, T 10% fall, dT 90-10%
Export Analysis Creates ASCII data files includes: time series, twitches, averages, statistics

Resources

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