Knowledge Base

Install DanioScope

Make sure that the USB hardware key is not plugged in your computer before installing the software.

1. Insert the DanioScope Installation USB stick into one of your computer's USB ports.
2. Browse to the USB stick contents and double-click Setup.exe.
3. In the Welcome screen, click Next>.
4. Read the license agreement and select I accept, then click Next>.
5. Click Install.
6. Click Finish to complete installation.

To Start DanioScope

1. Before you start DanioScope, insert your license key into a USB port of your computer.
2. To start DanioScope, on the Apps screen under Noldus choose DanioScope.

Your First DanioScope Experiment

Create a New Experiment

To create an experiment from scratch, choose File > New. Give a name you want to give to the experiment, and click Create.

• To create an experiment from a pre-existing template, choose File > New from Template. Select a template from the Template list.
• The default location is C:\Users\Public\Documents\Noldus\DanioScope\Experiments. To change the location for all the next experiments, choose File > Preferences.

The Experiment's Basic Properties

Measurements

Choose Setup > Experiment Settings.

Choose the measurements you want to take:

• Activity — To measure activity bursts of the embryo.
• Cardiology — To measure heart rate.
• Flow — To measure fluid movement rate in blood vessels or the gut.
• Morphology — To measure body parts and organs. Click the Add button to define additional measurements.

Independent Variables

You can associate your measurements with one or more variables important for analysis, named independent variables.

Choose Setup > Independent Variables. Two variables, Compound and Time Point, are predefined. Choose which variable you want to associate with measurements. Click Values to edit the possible variable values. To add a new variable, click the Add Variable button.

Record Media Files

• We advise to name video and image files in such a way they suggest which subject(s) and time points are recorded. This may be very useful when copying files from one computer to another.
• Test the format of your video before recording videos of your experiment. To test a video format, in the Acquisition screen load the video file and play it back. For the list of supported formats, see the Questions and Answers in the DanioScope Help.
• Automatic detection of embryos is only possible when the video is not distorted relative to the original aspect ratio of the camera image. If the aspect ratio of the video file is distorted, convert the video file to one of the formats compatible with DanioScope, and make sure to adjust the video aspect ratio.

Activity

Aim

To record and analyze bursts of movement in zebrafish embryos.

Procedure

1. Choose Acquisition > Activity.
2. Click the Add Videos button and load one or more video files.
3. Select a video file and position the video to the first frame where you want to start measuring.
4. Click the Detect Embryos button.
5. Select the values of the independent variables that apply for that video. Each detected embryo is marked with a circle and a code Sn / M1 (Subject/Measurement), where n is a progressive number.
6. Click the > button and repeat the procedure for the next video file.
7. Click the Acquire All button.

To Check Activity Data

Click one of the rows in the results table and click the Activity Settings button. In the window that appears, play the video and check that the Activity (%) peaks correspond to the actual behavior of the embryos. If necessary, adjust the Activity onset and Activity offset settings and click Apply.

• If DanioScope cannot find the embryos, you can define them manually. For details, see Add a measurement in the DanioScope Help.
• If an embryo shows spatial displacement in the video, for more accurate measurement of activity, you can only analyze the part of the video where the embryo stays in the same place. For this and other tips, see Questions and Answers in the DanioScope Help.

Cardiology

Aim

To analyze the heart area in zebrafish larvae, and estimate the heart rate.

Procedure

1. Choose Acquisition > Cardiology.
2. Click the Add Videos button and load one or more video files.
3. Select one of the video files in the list.
4. Choose one of the drawing tools and draw a shape around the heart of one of the subjects.
5. Enter the number of subjects in the current video and click Add.
6. Place a heart area over each heart in the image. Repeat the procedure for other videos.
7. Click the Acquire All button.

For more information, see Cardiology in the DanioScope Help.

Flow

Aim

To estimate flow of fluids in specific areas of interest, like a blood vessel or a gut section. Note: Flow does not give results in physical units like l/s.

Procedure

1. Choose Acquisition > Flow.
2. Click the Add Videos button and load one or more video files.
3. Select one of the video files in the list.
4. Choose a drawing tool and draw a shape over the blood vessel or gut in such a way that it fits exactly inside the vessel/gut section.
5. Release the mouse button and assign the shape to an existing subject S01, S02, etc., or add a new subject if you have drawn the shape on another larva in the image.
6. Repeat steps 4-5 for other areas, subjects and videos.
7. Click the Acquire All button.

For more information, see Flow in the DanioScope Help.

Source: EthoVision XT 18 - THC - Trial and Hardware Control, Noldus Information Technology