DanioScope is a software tool to investigate a scope of zebrafish embryo and larvae parameters. It includes features to measure embryo activity, cardiovascular measurements, morphology, and gut and blood flow. Measurements are based on video files and images, offering an easy-to-use and non-invasive way to study your zebrafish. Most importantly, DanioScope allows you to test many animals simultaneously, increasing the throughput of your research significantly, and ultimately saving you a lot of time when it comes to preparing your experiment, acquiring data, and analyzing the results. Moreover, automating your research with DanioScope means using a consistent measuring tool – eliminating inter- and intra-observer variability.
DanioScope software is available in combination with the ZEISS SteREO Discovery.V8 microscope to acquire crisp and detailed images and video files to use in the DanioScope software. For more information about this microscope, please visit www.zeiss.com.
Zebrafish embryos are very interesting for toxicology research because of the short time frame in which they develop into adults; the development of their internal organs starts to show in a matter of hours. Best of all, you can monitor it closely because they are completely transparent. Movements such as tail flicks, coilings, and convulsive behavior start as early as 17 hours after fertilization (hpf) and are important measurements in toxicology and pharmacology because they reflect the development of the nervous system and are highly susceptible to pharmacological manipulation.
With DanioScope you can use video files of one or multiple zebrafish embryos to study their activity. The software automatically recognizes the embryos and selects each embryo (in chorion) as a zone of interest in which the activity is automatically registered and measured. Of course you can also select the area(s) of interest in which you want to measure the activity yourself.
The software detects activity to measure the following parameters:
- Burst activity – percentage of time (from total measurement duration) the embryo was moving
- Inactivity – percentage of time of inactivity (100% - burst activity).
- Burst duration – total time spent active (sum of all movement durations)
- Inactivity duration – total time spent inactive
- Burst count – number of times the embryo moved
- Burst count / per minute – number of movements per minute
Compared to the manual method of visually counting the activity, DanioScope can save you 75% experiment time (average from beta testing*).
Depending on your medium, embryos might float to another location during the experiment. When this happens, you can redefine the position of the embryo and measure activity at this area for the selection duration of the video. Measurements for this embryo are averaged.
The heart is the first organ to develop in zebrafish, and development is similar to the human heart. The zebrafish cardiac system is assessed by measuring the heart rate or inter-beat interval. Traditional methods include manual counting (using a stopwatch), using a micro pressure system, Laser Doppler microscopy, or electrocardiograms. These methods are labor-intensive, time-consuming, and require specific training to perform.
In DanioScope, you can select one or more video files showing one or more larvae each. Then you indicate the heart area by drawing a rectangle around it for each larva. Within the area(s) of interest, activity is automatically monitored, and the number of beats per second (BPS) and beats per minute (BPM) are automatically calculated. With just the video of your larvae and DanioScope you can easily monitor the heartbeat; there is no need for fluorescence labelling or other techniques that require specific training. Beta testing* shows that DanioScope saves you at least half the time compared to manual methods.
In addition to heartbeat measurements, blood flow is another useful parameter in toxicology and other research areas. Malfunctioning of the heart might not always show in the blood flow measurements, and vice versa, depending on the cause, and so researcher often measure both.
Blood flow can be measured using hand counting methods that might also require fluorescence labeling techniques, but as mentioned above these methods are labor-intensive and time-consuming. DanioScope uses video analysis to measure the flow in a blood vessel. In addition it is also suited to measure gut flow in a similar fashion.
To measure flow, you select (a part of) the blood vessel or the part of the gut, from wall to wall, in your video image. A perfect circle defines the area in which flow is measured. Similarly to larvae heartbeat and embryo activity measurement, changes in pixels on a frame-by-frame basis is used to measure flow. The blood or gut flow is presented in a percentage (of activity within that area).
Besides dynamic measurements, DanioScope also allows you to easily monitor the morphology of your zebrafish. You can upload images or take snapshots from videos to monitor changes in morphology. With intuitive drawing tools you can define distances or areas you are interested in, and after calibration DanioScope will measure lengths and surfaces automatically. This way you can easily determine tail length, eye size, pericardial area, or any other measurement, because you can define your own. By images from different time point, you can easily monitor growth and malformations over time.