ANY-Maze 1

ANY-Maze v. 5.2
This directional guide was made for ANY-Maze v. 5.2. Some information in here may apply to
future updates, but may not be entirely applicable. This guide does not support versions of
ANY-Maze earlier than 5.2.

IMPORTANT
****If you are reinstalling ANY-Maze on another computer - m ake sure you deactivate
the program on the old computer before reinstalling. You may have to contact
ANY-Maze support before you do this. Be sure you have the deactivation code, other
wise you will not be able to reinstall ANY-Maze. Previous information such as the
license and serial number of ANY-Maze is located in the ANY-Maze CD case****

Renew ANY-Maze updates and support every 12 months. See ANY-Maze website for details.

ANY Maze is like Microsoft Word or Excel in that you cannot do anything useful without
first opening or creating a file
In ANY Maze, each file equates to a single experiment and so the system tends to refer to
experiments rather than to files
ANY Maze is structured around 5 principal pages called: Protocol, Experiment, Tests,
Results and Data.
You can switch to any page at almost any time, even while testing, simply by clicking the
pages title in the page bar at the top of the screen.
A novice user may want to click Getting Started section of the ANY Maze help and
support centre - access this through the help menu.
To ask a question, contact techsupport@anymaze.com
If you are more of a visual learner, feel free to watch InsideScientifics two part webinar,
Animal Behavior Video Tracking Using ANY-maze Software, located on YouTube
(https://youtu.be/dQU5yfiDg1Y)
 ANY-Maze 2

Table of Contents

1. Introduction..1
2. Table of Contents.........2
3. Basic Setup....3,4
4. Exploring Advanced Features 1........5,6
a. Three Point Tracking (& Brief Novel Object Recognition and Sociability) ...5
b. Heat Maps & Track Plots......6
c. Virtual Switches.....6
5. Exploring Advanced Features 2 ....7
a. Precise Zone Entries....7
6. Exploring Advanced Features 3.8,9
a. Moveable Zones & Procedures...8,9
7. Exploring Advanced Features 4..10
a. Sequences & Calculations....10
8. Taking ANY-Maze Beyond Video Tracking 1..11-14
a. Inputs & Outputs...11-14
9. Taking ANY-Maze Beyond Video Tracking 2...15
a. Scoring Behaviors Manually..15
10. Analyzing Results & Measuring Data.16-18
a. Statistical Analysis, Graphs, and Track Plots...16
b. Transferring Data to Other Programs..17
c. Analysing Results Over Time...17
d. Accessing Raw Data...18
 ANY-Maze 3

Basic Setup

1. Open ANY Maze

2. To start new experiment, click New Empty Experiment
a. This will bring you to the protocol page
b. You must set up ALL bolded items on the left hand side.
3. Click Add Item to add video.
a. If you have a camera set up to ANY Maze, select that instead.
b. Once uploaded, name your video source.
4. Click Add Item and then New Apparatus
a. Name your apparatus
5. Use the rectangle, circle, or line tool to draw and select the area you
wish to track.
a. ANY Maze will track ONLY what is in the lines of your
drawing.
b. Use the green ruler tool (see right) to tell ANY Maze how big
the apparatus is. Make sure you know your EXACT
measurements before doing this.
6. If you wish to divide the apparatus into a grid, click grid settings and
set your preferences. Each section in the grid can be selected as a
zone.
7. Click Add Item and then select New Zone
a. Name zone and area(s) you want to measure. (ex. if you wish
to track whenever the animal enters the corners, click all
corners you wish to measure, they do not have to be
touching)
b. You can create multiple zones, but you must repeat step 7 for each different zone
you wish to track. (ex. If you have corners selected but wish to also track when the
animal is in the center, repeat step 7 and select the center instead)
c. Click highlight zone when animal is in it
8. Click Animal Color on the left hand side and answer the question. This tells ANY Maze
what to track. ANY Maze will NOT work if this is not answered.
9. Click Stages on the left hand side.
a. A stage is ANY Mazes way of dividing up your experiment into different sets of
trials that you are going to perform.
b. You dont have to have multiple stages if you dont want to. The default is one
stage and one trial. You can easily edit this.
c. Tell ANY Maze how long the test will last. ANY Maze can track each video up to 7
days long.
10. To add what you would like to see in your results, look at the far left hand column on the
Protocol page. This will have this option under Analysis and Results. If you do not see
this, scroll down.
 ANY-Maze 4

a. You can then click on Results, reports, and data

b. under that tab there will be the Animal and test details reports. Click that.
c. Click on everything that you would like measured for your test.
d. Whatever you change in the protocol will also
affect the protocol for previous, current, and future
tests on the same file. ANY Maze will reanalyze
data so it includes everything you selected for all
tests.
11. You can now save this protocol as a protocol file and reuse
it whenever needed. Doing this, you would not need to set
up protocols every time. All you have to do is load the
protocol
12. Click on the Experiment header
a. Name your groups - this is next to the Treatment
column on the page. Add the number of animals in
the treatment. Repeat for however many
Treatment groups you want, i.e. Treatment 1, Treatment 2, etc.
b. After this, you are now ready to start performing tests
13. Click on the Tests header
a. On the left is your test report, showing how many tests you will perform and the
total number of animals from all of your groups, or
treatments.
14. Click Rewind and start video at test start under the small
dropdown button next to it next to the button.
15. Click the button to start the test.
16. You can switch to any page while the test is running.
However, there are some restrictions to this - you cannot
edit certain aspects of the test while it is running, (ex. you can not change the
definition of the zones) so make sure to double check that everything is correct before
starting
17. Once your test is finished running, point your cursor at the test number under the Test
column on the left of the screen and click. This will take you to the Test Details Report
18. The Test Details Report will show you the results.
a. If you would like to include more results than
what is shown, click on the light blue Results,
reports, and data underneath the Test
Results section.
b. Look back at Step #10 and follow the
directions.
c. If you return to the test page, it will include all
of your new selections.
 ANY-Maze 5

Exploring Advanced Features 1

Applicable to: Open Field, Object Recognition, Sociability, Home Cage, Three Point Tracking,
Heat Maps, Virtual Switches
No features in ANY Maze are apparatus specific

Three Point Tracking (& Brief Novel Object Recognition and Sociability)
1. Three point tracking is where you are not only tracking the animals center point, but
also the head and base of the tail.
2. Go to the Protocol tab and find the Tracking section on the far left hand column
a. Click Tracking the animals head and tail and answer the yes or no question.
3. You can then go under the Testing section on the Protocol page on that
same far left column, and click What to display while testing
a. You can then click mark the animals head and mark the animals
tail. You can also connect the animals points with lines.
4. Run the test again
5. This is useful for Novel Object Recognition.
a. You can put an object into the maze and then draw a zone on or around the
object, possibly leaving a few centimeters between the edge of the zone and the
edge of the object.
b. You can then ask ANY Maze how much time that animals head spends in that
zone, and that would be considered as investigation of the object.
c. This can also apply to Sociability testing.
6. You can then go back to Tests if you have already run the experiment and look at the test
results. Make sure you specified what you wanted the results to show before running the
test.
7. You can always go back and re-edit whatever you see fit after you have performed a test;
this includes adding or drawing new zones and whatever else is on the Protocol page.

CONTINUED
 ANY-Maze 6

Heat Maps & Track Plots

1. On the Tests tab, click Related Reports on the top bar of the screen, located
about midway.
a. This will give you a list of reports related to your current report
2. You can then click Tests track plot which displays a track plot or heat map of
the animals center or head position.
3. Once clicked, it will show you the animals track plot. You can then click Plot
Type at the same top bar as before
a. You can then select different options for the tracking location or change
it to a heat map.
b. NOTE that the heat map is not entirely accurate

Virtual Switches
Virtual switches do not have a particularly obvious name, so an example will be used
1. For speed, ANY Maze will track your animals around your maze by every picture it gets
from the cameras, which is about 30 positions per second.
a. It does not store the 30 positions per second, usually about 10, but you can
change that in the protocol.
i. This means you would have 600 positions for a 1 minute test, and ANY
Maze will use these to calculate how fast the animal is moving
ii. This will end up with 600 values for speed for each animal tested
2. Virtual switches takes these values and put them into a more analyzable form
3. To do this, go to the Protocol tab and click Add Item at the top bar.
a. Then, click New Virtual Switch
b. Name your switch
4. Find the bar that says -unspecified-, which should be about a third of the way down the
page. This tells ANY Maze when you would like the virtual switch to turn on.
a. Select what is appropriate for your test
5. Once selected, more options will show up, showing the list of possible measures that can
be chosen.
6. Then, at the bottom of the page, you can select certain values and limits to create
thresholds
7. Make sure you have selected what results you have wanted to display for this, otherwise
they will not appear. (see step 10 on Basic Setup, page 2).
 ANY-Maze 7

Exploring Advanced Features 2

Precise Zone Entries

Applicable to: Elevated Plus-Maze, Zero-Maze, T-Maze, Y-Maze, etc.

1. Following the same protocol as the Basic Setup (see page 3), we will examine the zones
we have set up.
2. Each Zone has an Option called Zone Entry Settings
a. This determines when an animal is actually in a
particular zone.
b. You can choose the center (default), head (good for
novel object recognition), or entire body of the
animal.
c. For the entire body, you can input what percentage of
the animal you consider precise zone entries.
i. 80-85% of the animal is
usually considered 4 paws in the zone.
3. ANY Maze will consider even a slight change in the 80-85% zone entry the animal makes
either an entry or an exit. For example, if the
animal moves back to 84% of its body in the zone,
ANY Maze counts this as an exit. To human
observers, this would not be correct. To fix this, we
must set up parameters.
a. 75% is the example given (image right). This
means at least 75% of the animal must be in
the zone for it to count as an entry. Once
this value is below 75%, it is now an exit.
Once more, this value can be changed.
b. A 10% difference between the minimum
entry and exit is seen to be ideal.
c. Remember that it is always good to double
check that all entries and exits are correct.
Hand scoring and double checking results is extremely important.
 ANY-Maze 8

Exploring Advanced Features 3

Moveable Zones & Procedures

Applicable to: Morris Water-Maze, Radial Arm Maze, Y-Maze, T-Maze
1. We will be looking at moving platform zones in a water maze as an example. Following
the same protocol as the Basic Setup (see page 3), we will examine the zones we have set
up.
2. Add a new zone and name it
3. The list box (see right) has a default choice of Position of the zone remains the same in
all tests. This can be changed.
4. We can select Position varies within (and possibly
between) the animals This means that between
animals and between different trials the position of
the zone could move.
a. We now have to tell ANY Maze what
potential positions these are
5. Click Add Item and Add Zone Position
a. Name your zone position. Cardinal directions
are common names.
b. Do this as many times as needed.
6. Click Add Item and New Procedure
a. Name the procedure
7. Create the procedure by dragging statements over to the far right box. In this example,
we are using the Wait Until statement
 ANY-Maze 9

8. Once Statements have been dragged over, click on the Events tab. This tells ANY Maze
what it is waiting for. In this example, it is waiting for an event related to zones. Click the
zones option, specifically the platform zone. ANY-Maze reads this code from top to
bottom - make sure it
is in the correct order
you want.
9. For this example, we are
interested in the Animal
enters the platform zone
a. Click the purple
Animal enters the
platform zone
button and drag it
over, releasing it on
the Wait until
button. These two
should then combine
10. Go back to Statements
a. Click and drag the green Action button underneath the Wait until buttons.
11. Go to the Actions tab at the top. This is next to
the Events tab.
a. Click and drag the End the test button
onto the Actions button.
b. This tells ANY Maze that the test will end
once the animal enters the platform zone,
however, it needs a reason to end it.
12. Click the pencil on the actions button.
a. A box will pop up saying Create Test and
Reason
b. The name will be the name of the procedure by
default.
c. We enter a reason for the procedure ending because ANY Maze will give you an
analysis as to why the test ended. If the animal found the platform, it will explain
that it ended there because of that reason. However, if the animal did not find the
platform and the time of the test ran out, it will give that
reason in the results and analysis.
13. When the Test is about to be performed, go to the Tests tab. At the
top of the video, there will be a row of text.
a. This will ask you to click on the platform zones position.
b. Tests will not start unless you give the whereabouts of the
platforms position.
14. Remember to always click the Rewind and start video at test start
option next to the play button before performing the test.
 ANY-Maze 10

Exploring Advanced Features 4

Sequences & Calculations

Applicable to: Y-Maze, T-Maze, Radial Arm Maze
In this example, we will be sequencing and calculating spontaneous alternations. Spontaneous
alternations is a behavioral test for measuring the willingness of animals to explore new environments.
Rodents typically prefer to investigate a new arm of the maze rather than returning to one that was
previously visited. Every time this happens, we count this as an spontaneous alternation. ANY Maze can
turn this into a percentage. For a Y-Maze, each arm of the maze can be named A, B, and C, or 1, 2, or 3,
depending on your preference. For these tests, you may want to use the Precise Zone Entries on page 7.

1. Click on the Protocol tab and click sequences on the far right column
2. To calculate spontaneous alternations, scroll down to the calculations tab, this will be at
the bottom
3. Click Add Item and Add Calculation
a. You will be asked to name and then input an equation to find your calculation,
make sure you are extremely
specific and use parentheses.
Here is the equation for
spontaneous alterations on a Y
Maze (right)
4. Scroll down to Analysis and Results and click Animal and Test Detail
Reports
a. Scroll down to the bottom to Calculation Results and click the
calculation you just created.
5. To add a sequence, click Add Item and select New Sequence
a. Name your sequence. Be sure to create and name all possible sequence
combinations separately.
b. For a Y-Maze, you would have 6 different sequences ex. ABC, ACB, etc.
6. Click Add Item and then New Sequence Step
a. This is where we will select the zones for entry. If we start off with
ABC, we input A where there is a selection that says The following
zone.
b. Create a step for every arm the maze has for each sequence, so for
sequence ABC, step 1 is A, step 2 is B, and step 3 is C
7. Repeat for each sequence you have. Be sure there arent any repeats.
8. You can now run your test. Once finished, you can look at your results.
 ANY-Maze 11

Taking ANY-Maze Beyond Video Tracking 1

Inputs & Outputs

In this sense, an input is something like a lever an animal can press or a photobeam it could break. An
output could be something like a lamp that we could turn on or a shocker to give an aversive stimulus to
an animal. Outputs typically need to be controlled, which will bring us back to the procedures on page 8.
In this tutorial, we will be using levers, switches, pellet dispensers, and shockers as examples.

1. Open your experiment.

2. First, change the protocol mode. To do this,
go to the Protocol tab and look at the first list
box.
a. The default choice is video tracking
mode. Click the box.
b. In the Standard Modes section, select Video Tracking Mode with
Input/Output
3. Set up your experiment normally. See the Basic Setup section on
pages 3 & 4 for reference.
4. Go back to the Protocol when finished.
5. Select Add Item and then New Input Item. Select New
on/off input This is used for an on/off switch.
6. Scroll down to Keys and I/O on the far left hand column.
a. Under On/Off Inputs there should be your new switch.
b. Name your switch and select what would trigger this
input
7. To create an output, select Add Item and select New
Output Item, then New Output Switch
a. Output switches can be used for things such as a pellet
dispenser
8. Name your output switch.
9. Select what the output switch should do when activated. In this case, we
will be using Pulse the Output on the Following Duration for whatever
amount of time needed
10. We can add a shocker into our protocol too by going to Add Item,
New Output Item, then New Shocker
a. Name your shocker.
b. Specify the duration of your shock and the intensity of the shock in
milliamps. The example to the right is exaggerated - do not
shock your animal for this long
11. We now need ANY-Maze to understand the logic of inputs/outputs by
using procedures.
 ANY-Maze 12

12. Scroll down to the Testing section on the Protocol page, and select
Procedures
a. Set up your procedures. Look at page 8 for reference.
b. In this example, we will have 2 procedures, pictured right
c. Building procedures are much like lego blocks - here is the
example of the lever press below.

13. In the picture above, take note that the light orange repeat block wraps around the light
red wait until and the pink set blocks. This is crucial to having the procedure work
correctly. Statements should always be wrapped around events, actions, and variables.
a. In the green lever press count block, you can see there is a + and then a
number. All this does is simply adds up how many time the animal presses the
lever in increments of one.
14. The pink block that says lever press count is called a variable. To add a variable, click
the X Create Variable button at the top of the page.
15. Another example is the pellet dispenser zone entry procedure, pictured on the next page
 ANY-Maze 13

16. In this example, we see the light blue If block. What it is saying is if the lever press
count is zero, meaning that the animal did not press the lever, then activate the shock
floor. Otherwise, or else, meaning that the animal did press it more than zero times,
activate the pellet dispenser.
a. The last pink set block says to reset that number to zero, negating whether the
lever was pressed or not, and then the process repeats all over again, thanks to
the repeat/forever block that encloses it.
17. We are now done telling ANY-Maze the logic it needs to run the test and the procedures.
Now, we can run the test and if done correctly, ANY-Maze will now apply these
procedures to the test.
a. You can tell if ANY-Maze understood your procedure if the animal enters
the zone and the names of the protocols now show up where and when
you wanted it.
18. When testing is done, you can now look at your results.
19. More information on Keys and I/O
a. Signal inputs are typically a voltage coming from another piece of an
apparatus (ex. detecting an animals heart rate or core temperature.) This would
come into ANY-Maze as a signal, allowing you to see that
information.
b. Sensors - ANY-Maze supports 4 different types of sensors:
temperature, humidity, weight, and light.
c. Rotary Encoders - detects when something rotates and counts how
many rotations i.e. a running wheel
d. Speakers - ANY-Maze can play any sound file that is on your
computer, even white noise. Can control volume too
e. Analog outputs - voltage coming out of computer. This would be a
voltage that is being controlled by what an animal may be doing
 ANY-Maze 14

(ex. a voltage that grows/shrinks as an animal runs across certain parts of the
maze)
f. Temperature and lighting controllers - self explanatory. Hotter/colder,
dimmer/brighter.
g. Syringe pumps - allows you to control flow rate, duration of when the syringe
pump is on, direction to withdraw, etc.
20. All of these can easily be controlled with these procedures. Some of these may need
interface devices to be connected to a computer. ANY-Maze sells some interface devices
called AMY devices, but they also support some 3rd party devices as well.
a. Most syringe pumps do not need any additional interfaces, but ANY-Maze needs
to be able to detect it.

CONTINUED
 ANY-Maze 15

Taking ANY-Maze Beyond Video Tracking 2

Scoring Behaviors Manually

For example, ANY-Maze cannot determine when the animal is grooming, but this is a behavior you would
like to measure. You can manually score this on ANY-Maze

1. Create your experiment or continue it in ANY-Maze.

2. Scroll down the far left column and select Keys from the Keys and I/O section in
the Protocol.
3. Select Add Item, then New Key
a. Name your key and specify which key on your
keyboard you would like to press to score the
behavior
4. Be sure to check your What to display while testing tab
underneath the Testing section on the Protocol page and make sure that list any active
keys is selected
5. Make sure the Animal and test details results under the Analysis and Results section of
the Protocol page also has your key results selected - this will be shown as the key name.
6. You can now run your test. If you are measuring how long the animal is performing an
action, such as grooming, hold down the key from start to stop.
7. If you have multiple keys you need to press for different behaviors, such as rearing,
sniffing, fighting, etc. we will follow a slightly different process.
a. Create however many keys you need like
before.
b. Now, we will be changing the Protocol
mode, like we did in the Input & Output
section on page 11. This time, we will be changing it to
TakeNote Video Observation Mode. This does not track
the animal, it only allows the user to observe it.
c. There are 3 types of TakeNote Observation Modes, choose
whatever best suits your experiment
d. With TakeNote Video Observation Mode, ANY-Maze will not run the test
automatically, so you must do it manually. Be sure to rewind the video before
starting.
e. Remember to press the key for as long as the animal is performing that action if
you are measuring time.
f. If you dont want/cannot press multiple keys at once, you can
mark the test for additional observations. Press the Mark Test
for Additional Scoring at the top of the page and perform the test again for the
other observations. ANY-Maze will add on the new scoring to the results.
g. There is no limit for how many keys or observations to measure.
 ANY-Maze 16

Analyzing Results & Measuring Data

Statistical Analysis, Graphs, and Track Plots

We will now be moving onto a new tab, the Results tab.

1. Once testing is complete, click on the Results tab at the top of the page for more
detailed results.
2. The Results to Include section will give you options to select so ANY-Maze will
include that data in the report.
3. You can also group the data under the Data Grouping section. There are many options
to choose from, ranging from treatments, trial, sex, and more.
4. Under that section there is Report Format. This will have
options on what you would like to include in the report.
5. Below that, there is Filter tests This is where you can filter
the tests that you actually want analyzed.
6. If you select View the Report at the top of the page, it will show you the test duration
and analysis between different groups.
7. There are different result styles you can look at to get a better feel of your
results. This is located at the top of the page.
8. The Graph style will show you a graph, the Statistical style will show
you all the numbers and statistics, and finally, the Track Plot style will
a. Text and Graph are very similar to the first page on the Results
tab, with the data grouping, report format, etc.
i. Statistical is a bit different, asking you about independent and
dependent variables to analyze.
ii. ANY-Maze will apply the most appropriate test for the data you wish to
use.
iii. You can also choose which post hoc test using ANOVA (analysis of
variance) you would like to use.
b. Track Plot report asks you what sort of plot you would like to see, how they are
grouped, and how you want them to be filtered.
9. You can then click View the Report when satisfied.
10. Everything seen in the Report can be
saved by right clicking or screenshotted
to include in papers, posters, and more.
a. You can copy the plot to your
clipboard and it will be saved in
a vector format, meaning it will
stay crisp and sharp when
resized.
 ANY-Maze 17

Transferring Data to Other Programs

1. Go to the Data tab at the top bar. This will show up as one large spreadsheet.
2. You can choose what columns you want included in the transfer. To do this, click Select
Data
a. You can now check the boxes with the information that you would like to
include.
b. Select what you would like by highlighting all information you would
like to save with your mouse.
3. You can now copy the spreadsheet, save it as a file, or send it by email
4. These selections can be copied and pasted into Excel or SPSS.

Analysing Results Over Time

1. Go to the Protocol tab and scroll down to the Analysis and Results section.
2. Click on the Analysis Across Time option
a. Here you can specify the
duration of time segments
b. This chops the experiment
into intervals.
3. Go to your Results page
a. Click the Data Grouping
option and Group data
by, then select Segment
of test
4. The results will now be split up by time intervals.
5. You can look at the graphing tab to see the
consistency of the intervals (see right)
6. We can also split this up by time periods. This
essentially takes a chunk of your test and gives you
the result from that chunk.
a. To do this, go back to the Protocol and to
Analysis Across Time once more.
b. Click Add Item and New Time Period
c. Name your time period and specify when you want to analyze and end your
results.
 ANY-Maze 18

Accessing Raw Data

1. Go to the Protocol page and scroll down until you find Analysis and Results,
then select Results, Reports, and Data and then select its sub category, Test
Data Report
a. Select all the data you would like to be measured and shown in the
data report.
2. Go to the Tests tab, click Test Schedule Report, and then click the number
of the animal you would like to look at. Select Related Reports, then Test
Data Reports
a. This will show you the x and y coordinates at different time periods
3. You can then select, copy, email, or save the data.
4. To save several individual reports or all of them at once, go to the File tab in
the top left corner and select Data Tools. You have two exporting options
(pictured left)
a. The first option is Output Experiment as XML, where you can
export all the information in one place.
i. You can choose what information you would
like to export. This can be imported into systems such as MatLab, Excel,
and more.
b. The second option, Export Test Data will create as many files as you have tests
in your experiment, containing all the information needed.