Water-maze example protocol
This protocol is part of an example water-maze experiment for ANY-maze.
Under this protocol the experiment's treatment groups will be coded in the order in which they are entered (i.e. the first treatment group will be A, the second will be B etc.) and the animals will be assigned to the treatment groups in a 'cyclic' order. This will mean that the first animal will be assigned to the first treatment group, the next animal will be assigned to the next group, the third animal to the third group and so on until one animal has been assigned to each group. Then the process will 'cycle round' and the next animal will be assigned to the first group again, the second to the second and so on. For example, in an experiment with 3 groups of 4 animals this would mean that animals would be assigned as follows: A,B,C,A,B,C,A,B,C,A,B,C.
Video sources
| Water-maze | Images for this source are captured from the Water-maze example image interface. The source uses the entire captured image which measures 464 x 452 pixels. |
Apparatus
| Water-maze | The image of this apparatus is provided by the Water-maze video source. Figure 1 shows the apparatus map for this apparatus, the 'ruler' line shown in the map represents a real-world distance of 1500 millimetres. The user can control tests in this piece of apparatus using the computer's F5 function key. |
Figure 1. Water-maze apparatus map.
Zones
| NW | Figure 2 shows the area(s) in the apparatus which correspond to this zone. The system will consider that the animal is in this zone whenever its centre point is within the zone. While tracking the system will highlight this zone whenever it considers that the animal is within it. |
Figure 2. Position of the NW zone in the Water-maze.
| SW | Figure 3 shows the area(s) in the apparatus which correspond to this zone. The system will consider that the animal is in this zone whenever its centre point is within the zone. While tracking the system will highlight this zone whenever it considers that the animal is within it. |
Figure 3. Position of the SW zone in the Water-maze.
| NE | Figure 4 shows the area(s) in the apparatus which correspond to this zone. The system will consider that the animal is in this zone whenever its centre point is within the zone. While tracking the system will highlight this zone whenever it considers that the animal is within it. |
Figure 4. Position of the NE zone in the Water-maze.
| SE | Figure 5 shows the area(s) in the apparatus which correspond to this zone. The system will consider that the animal is in this zone whenever its centre point is within the zone. While tracking the system will highlight this zone whenever it considers that the animal is within it. |
Figure 5. Position of the SE zone in the Water-maze.
| Island | Figure 6 shows the area(s) in the apparatus which correspond to this zone. The system will consider that the animal is in this zone whenever its centre point is within the zone. While tracking the system will highlight this zone whenever it considers that the animal is within it. |
Figure 6. Position of the Island zone in the Water-maze.
Points
There are no points defined in this protocol.
Sequences
There are no sequences defined in this protocol.
Keys
There are no keys defined in this protocol.
Switches
There are no switches defined in this protocol.
Test settings and options
| Animal colour | The animals that will be tracked are lighter than the background of the apparatus. |
| Tracking the animal's head | The system will not attempt to track the animal's head. |
| Immobility detection | The system won't detect immobility while tracking the animal. |
| Automatic starting of tests | The system will automatically start tests when the experimenter leaves the camera's field of view. |
| What to record while testing | The system will record the animal's position at most 8 times per second. |
| What to display while testing | While tracking the system will display the time remaining in the test. It will also mark the position of the animal's centre of gravity. |
An indicator will be shown next to the position of the animal when:
| • | A 'key' activity is occurring |
| Advanced tracking options | The apparatus is not inside an enclosure. The animals will \i1 not \i0 be in the apparatus before the start of the test. |
Fields
| Sex | This field is used to record data for the animals and will accept any one of the following entries: Male,Female. |
| Weight | This field is used to record data for the animals and will accept a numeric entry in units of g. |
Stages
| Training | Test duration |
| Tests in this stage have a duration of 120 seconds (although tests may end before this if a test end action is triggered). |
| Number of trials |
| The stage will end after a maximum of 6 trials. The trials for an individual animal will end before this if the animal satisfies the following rule: |
| • | Entered island within 30s in 2 consecutive trials : The stage should end if the animal enters the Island zone within 30 seconds of the start of the test, in 2 consecutive trials. |
| Test order |
| In this stage the animals will be placed into groups of 2 and each group will be tested in all of its trials before the next group starts the stage. |
| Treated | Test duration |
| Tests in this stage have a duration of 120 seconds (although tests may end before this if a test end action is triggered). |
| Number of trials |
| The stage will end after each animal has had one trial. |
Events and actions
| Island entry | The event will be triggered if the animal enters the Island zone. When triggered this event will automatically be reset after all the effects of its activation have been processed. The event will only be processed in the following stages : |
| • | Training |
| • | Treated |
| End test | This action will occur when any of the following events becomes active : |
| • | Island entry |
The effect of this action is to :
| • | End the test but allow the user to force it to continue (if possible) |
Calculations
Number training trials
| This calculation is performed as follows: |
| Count { Test duration, Training : Trial 1 - Training : Last trial } |
| The result of this calculation will be quoted to 0 decimal places. |
Analysis
ANY-maze will use a level of significance of p ≤ 0.05 in all statistical tests.
In any analysis that the system performs within tests (i.e. through the use of the Segment measure) it will use a segment length of 30 seconds. That is to say that it will break the tests into equal length segments of this duration (although the last segment may be shorter) and analyse the results in each one.
In this protocol the test duration will be used as the latency for events which don't occur. For example, if the animal does not enter a particular zone during a test then the latency to the first entry to the zone will be the test's duration. This will apply to: Zone entries/exits which don't occur, key presses which don't occur and sequences which don't occur.
When calculating averages where the denominator of the calculation is zero (for example, the average speed in a zone that the animal never entered) the system will consider the result to be undefined. This will mean that, in these cases, the animal won't be included in the results for the measure which will cause the measure's N to be reduced.
Reports
| Test schedule report | This report will include columns for the following data : |
| • | Test number |
| • | Animal number |
| • | Treatment code |
| • | Stage |
| • | Trial number |
| • | The reason for test end |
| • | Status |
Animal and test details reports
| These reports will show results for the following measures : |
| • | Test duration |
| • | Total distance travelled |
| • | Overall average speed |
Standard result
| Limits |
| All the sections of this report will only include analysis of tests which match all the limits listed below: |
| • | Trial = Training last trial or Treated trial |
| Statistics section |
| This section will show the results of statistical analysis of the following measures: |
| • | Latency to first entry to the Island zone |
| The analysis will use Stage and Treatment as independent variables and will be performed using a 2-way ANOVA (see note 1). Where necessary this will be followed by post-hoc analysis using Student-Newman-Keuls test . |
| The section will also show the groups' Ns, means and standard deviations. |
| Note 1: Where necessary tests for repeated measures will be used. |
| Graph section |
| This section will contain column graphs showing the results of the following measures: |
| • | Latency to first entry to the Island zone |
| The columns will represent the mean values of the measures with error bars of standard deviation. The x-axis will show Stage and the graph(s) will include a different series for each Treatment. |
| Data page | The Data page will include columns for the following data : |
| • | Animal number |
| • | Treatment |
| • | Treatment code |
| • | Stage |
| • | Trial number |
| • | The reason for test end |
| • | Test time |
| • | Test date |
| • | Total distance travelled |
| • | Latency to first entry to the Island zone |