The EZM is a circular, elevated track divided into quarters, with 50 cm diameter, 20 cm walls on the closed arms, and a 1cm rim on the open arms. Mice are gently placed onto the center of one open arm and allowed to freely explore for 10 minutes in normal room lighting. The experimenter leaves the room after placing animals on the apparatus. All trials are video-recorded in top-view, analyzed with Any-Maze tracking software, and results confirmed with blinded manual scoring. The time spent in and entries into the open arms as totals and per minute of the test, average duration of entry into the open arms, and time spent in the open arms across time of day tested were measured.
Tracking software
Manufactured by ANY-maze
Sourced in United States
Any-Maze is a tracking software that captures and analyzes animal movement and behavior data. It provides automated tracking and video analysis capabilities for a variety of research applications. The software can be used with various hardware setups to track the location and movement of animals in real-time.
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Lab products found in correlation
13 protocols using tracking software
1
Elevated Zero Maze Behavioral Test
2
Open Field Assay for Anxiety-Like Behavior
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Anxiety-like behavior was assessed using an open field assay as described in Besson and Martin (2005) [43 (link)]. Briefly, flies were placed individually in a custom-made arena (4 × 4 cm and 3.5 mm high), and their behavior video recorded during 10 min. The ANY-maze tracking software (http://www.anymaze.co.uk/ ) was then used to measure average locomotion speed, total distance traveled, number of entries to the central zone of the arena, and time spent in the central vs. the peripheral zone of the arena.
3
Spatial Memory Assessment in Rodents
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The testing (probe trial) was conducted 24 h after the last training session to assess memory. In probe trial, each animal was given 2 min to explore the pool. The spatial memory was assessed using the parameters, escape latency (time taken to reach the former platform area), number of times the animals crossed the annulus (former platform area), and time spent in the platform quadrant (the quadrant which housed the platform during the training).
To record the behavior of the animals during training and testing phases, a webcam (Logitech) was installed above the pool. The path followed by the animals in the pool was tracked using ANYMAZE tracking software (ANYMAZE). The videos were collected in a hard disk and analyzed manually.
To record the behavior of the animals during training and testing phases, a webcam (Logitech) was installed above the pool. The path followed by the animals in the pool was tracked using ANYMAZE tracking software (ANYMAZE). The videos were collected in a hard disk and analyzed manually.
4
Porsolt Forced Swim Test for Evaluating Antidepressant Efficacy
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The Porsolt forced swim test is a behavioral paradigm often used to evaluate the reversal of passive coping behavior by antidepressant therapies (Porsolt et al., 2001 (link)). The increased swimming behavior observed in this assay can be reversed by pretreatment with anxiolytic agents (Lee et al., 2017a (link)) and therefore, we incorporated it as an additional measure of negative affect. In this assay, mice were placed into a cylindrical glass container (11 cm in diameter) filled with room temperature water for 6 min. Using the AnyMaze tracking software, we measured the latency to the first immobile episode, the total time the animal was immobile, and the number of immobile episodes. Following completion of the test, the mice were returned to their home cages and were monitored until they were dry before returning to the colony room.
5
Anxiety-like Behavior in Mutant Mice
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The light/dark shuttle box test compared the photophobic/agoraphobic behavior between Homer2AA/AA and WT mice. The two lines of GRM5 mutant mice were also not available at the time of study and, thus, were not assayed in this paradigm. Mice were placed into a polycarbonate box (46 cm long × 24 cm high × 22 cm wide) containing distinct open (light) and closed (dark) environments for a 15-min trial. These two environments were separated by a central divider with an opening. Mice were first placed on the dark side, and the latency to enter the light side, number of light-side entries, and total time spent in the light-side of the shuttle box were recorded using ANY-Maze tracking software (RRID:SCR_014289 ). An increase in latency to enter the light, uncovered, side was interpreted as an index of anxiety-like behavior. The data were analyzed using a Genotype × Sex ANOVA.
6
Elevated Plus Maze Exploration in Mice
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The EPM assesses the natural conflict between the tendency of mice to explore a novel environment and their avoidance of a brightly lit, elevated, and open area (Montgomery, 1955 (link)). Methods have been described previously (Cheng et al., 2013 (link); Cheng et al., 2014b (link)). The ‘+’ apparatus consisted of two alternate open arms (35 cm × 6 cm; without side walls) and two alternate enclosed arms (35 cm × 6 cm; height of enclosing walls 28 cm) connected by a central platform (6 cm × 6 cm), elevated 70 cm above the floor. Mice were placed at the centre of the ‘+’ of the grey PVC plus maze, facing an enclosed arm, and were allowed to explore the maze for 5 min. The time spent on open arms, entries into the open arms, and the distance travelled on the open and enclosed arms were recorded by AnyMaze™ tracking software.
7
Optogenetic Modulation of Grooming Behavior
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Animals were placed in a two-chamber Plexiglas box; chambers were divided by an opaque black Plexiglas sheet with a 5.1 × 5.1 cm opening at the bottom that acted as a passageway between the two chambers. The LED floor was constructed using 595 nm and 460 nm LED strip lights (595 nm: 278 µW, 460 nm: 75.2 µW; Environmental Lights, San Diego, CA). Each box was equipped with a fan on the right side and lights on each side of the box. Mice were acclimated to the box for 10 min and then the floor diodes were turned on for 30 min. Movement was recorded and analyzed by ANY-maze tracking software (ANY-maze, Wood Dale, IL). K14-ChR animal videos were subsequently analyzed by a blinded experimenter who recorded the amount of time each animal spent grooming in the individual chambers.
8
Anxiety-like Behavior Assessment in Mice
9
Elevated Zero Maze Test for Anxiety
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The elevated zero maze test was conducted as previously reported [12 (link)]. Each mouse was allowed to explore the elevated zero maze for 5 min, and behavior was recorded using a wall-mounted Stoelting USB camera (Wood Dale, IL, USA). Behavior was analyzed in real-time by AnyMaze tracking software (Wood Dale, IL, USA). The percent time that the animal spent in the open quadrants (open zone occupancy) and number of entries into the open quadrants (open zone entries) were used as measures of anxiety-like behavior. Increases in open zone occupancy and entries were interpreted as a reduction of anxiety-like behavior.
10
Open Field Exploration Behavior
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The OF test was performed using a black, wooden box (60 × 60 × 25 cm) divided into 16 equal squares (15 × 15 cm). The four inner squares constituted the inner zone (30 × 30 cm), and the remaining squares (each stretching 15 cm against the walls of the apparatus) formed the outer zone. The animals were placed in the center of the box and allowed to freely explore the apparatus for 10 min under dimly lit (20 lx) conditions. The entries into the inner zone, time spent in the inner zone, and locomotor activity (measured as a traveled distance) were recorded by a camera above the apparatus and automatically scored by Any-maze® tracking software. The apparatus was cleaned with 70% ethanol and dried after each testing trial.
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