Knowledge Base

Data Management

Template Experiments

EthoVision contains a user-guided set-up with templates for several test paradigms (water maze, plus maze, 96-well plate, etc.). Many basic settings are pre-set, such as arena definition, detection settings, and parameters for analysis.

DanioVision Experiments

If you use DanioVision™, a system specifically designed for observing and analyzing zebrafish larvae behavior, you can select the DanioVision experiment option. The automated arena detection detects individual wells in standardized well-plates automatically and optimizes detection with a few mouse clicks.

Good Laboratory Practice

For those conducting Quality Assurance experiments, particularly in environments adhering to Good Laboratory Practice (GLP) standards, EthoVision offers specialized options to meet these rigorous requirements. This ensures that your experiments not only yield reliable data but also comply with industry standards for quality and reproducibility.

Flexibility

For researchers seeking maximum flexibility in their experimental design, the "New from scratch" option allows for complete customization of the experiment setup. This feature is ideal for innovative research that goes beyond standard protocols.

Setup

Experiment Options

• Tracking can be done live or from a video file.
• Use digital or analog cameras (but not a combination).
• Connect external data acquisition systems.
• Track in up to 99 arenas with up to 16 subjects per arena.
• Center point, center of gravity, nose and tail base tracking; color or color marker tracking.
• Activity analysis – a non-tracking option analyzing overall activity per arena based on pixel changes.
• Distance in mm, cm, m, or inches. Time in ms, s, min, or hours. Rotation in degrees, radians, gradians, or rotations.

Manual Scoring

The Manual Event Recorder is a built-in coding system which allows you to define a list of behaviors that you can score manually during and after data acquisition (live or offline).

• Start-stop behaviors – behaviors that are independent from each other.
• Point events – behaviors without a duration.
• Mutually exclusive behaviors – activating one automatically stops the other.

Contour-based Body Point Detection

• Any species - Default – Analyzes the contour to assign nose-point and tail-base.
• Adult Fish - For occlusions – Track zebrafish and similar fish when viewed from above.
• All species - For occlusions – No shape assumptions; for insects, crustaceans, large animals, or side-view fish tracking (center point only).

Contour adjustments: Erosion (decrease 1-10 pixels) and Dilation (increase 1-10 pixels).

Detection Settings

• DanioVision – Automatic detection method tailored specifically to larvae.
• Gray scaling – Pixels with gray value between thresholds as possible object.
• Static subtraction – Differences between reference image and current image.
• Dynamic subtraction – Like static, but updates reference on each sample.
• Differencing – Statistical comparison using variance in contrast.

Trial List

Trials organized in a cross table with system and user-defined variables. Schedule trials based on pre-recorded video files for batch acquisition. Import trials from previous experiments.

Arena Settings

Define arenas, zones, cumulative zones, hidden zones, overlapping zone layers, and points of interest. Unlimited zones per arena, any shape. Calibration enables velocity and distance calculations.

Multiple Arena Definition

For multi-well plates or groups of arenas, use the multiple arena set-up functionality. Rotate, scale, transpose, resize, and reposition arenas simultaneously. In DanioVision experiments, automated arena detection detects wells automatically.

Arena/Zone Auto-subdivision

Automated subdivision of circular and rectangular arenas into multiple equal zones with auto-labeling.

Multi Camera Set-up

Combine input from multiple cameras to track from multiple arenas without placing them all under one camera.

Trial Control Options

Program EthoVision to automatically start and stop tracking based on parameters. External command options can start other programs when conditions become true. With Trial & Hardware Control, define complete test protocols with real-time analysis. Conditions are applied to each arena individually.

Tracking Methods (Body Point Estimate)

The tracking method specifies how body points are estimated once the subject is detected. EthoVision offers the following acquisition methods:

1. Acquire data live – Live tracking requires no disk space for video storage, but creates no video backup either. Maximum trial duration (as tested) is 72 hours.
2. Acquire data live and record video – Live tracking while EthoVision records a backup video to an MPEG4 file. Maximum trial duration (as tested) is 72 hours depending on hardware.
3. Longer trials – To acquire trials longer than mentioned above, you can split your multi-day testing into multiple trials.
4. Record video and acquire data – EthoVision records a video to an MPEG4 file for data acquisition later. (Recommended if the computer is not fast enough for live acquisition and recording simultaneously.)
5. Acquire data from existing video files – Track data from video recorded with programs other than EthoVision.
6. Acquire data from a batch of existing video files at once – Track data from a series of pre-recorded video files.

In options 2–5, the video file is available for post-acquisition visualization. If you choose option 3, 4, or 5, it is possible to analyze samples at a rate faster than the actual sample rate. EthoVision supports MPEG2, MPEG4 and H.264 AVC video formats.

Series of Trials from Live Video Feed

You can use a live video feed to acquire a series of trials. You can predefine start & stop conditions and an inter-trial interval.

Data Acquisition

Acquire Additional Behavioral Data

You can score behaviors manually, adding scores live (while the test is running) or offline (by reviewing the video file). In addition, EthoVision is able to automatically recognize ten rat or mouse behaviors.

Acquire Data with DAQ System

External data acquired with a separate DAQ system can be synchronized with the tracking data. When you simultaneously acquire tracking and physiological data, EthoVision sends out a synchronization signal to the external DAQ system. After acquisition, you can import the physiological data and the two data streams are synchronized.

EthoVision offers import profiles for a number of DAQ systems, such as DSI's Ponemah. You can import signals sampled at a constant rate.

Source: EthoVision XT 19 Technical Specifications - Fish, Noldus Information Technology