ConductVision Zebrafish Package
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The ConductVision license for zebrafish tests.
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Mirror Biting Pham
SKU ME-4852
$1,490.00
IncludesStandard care
Specialized trapezoidal tank system with integrated mirror for quantifying aggression, boldness, and social behaviors in zebrafish and other fish species through controlled mirror-image stimulation protocols.
Scientist guidance
Louise Corscadden, PhD
Director of Science · ConductScience
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Key Specifications
Full details →- Model fit
- Fish, Zebrafish
- SKU family
- ME-4852
- Sizing
- Zebrafish: Height 15 cm · Width 7 cm
- Ordering
- Online checkout and quote request available
- Category
- Behavioral Mazes
- Build notes
- Confirm accessories, station layout, and support needs before purchase
The Mirror Biting Test Apparatus (Pham design) is a specialized behavioral testing system for quantifying social behaviors in fish species, particularly zebrafish. The apparatus consists of a 1.5-liter trapezoidal tank with dimensions of 23 cm base length extending to 28 cm mouth length, 7 cm width, and 15 cm height, with an integrated 15 x 7 cm mirror. This design enables controlled assessment of aggression, boldness, and social interaction behaviors through mirror-image stimulation protocols.
The apparatus employs defined behavioral zones with a contact zone within 0.5 cm of the mirror surface and an approach zone within 3 cm, allowing precise quantification of mirror-directed behaviors. Following minimum 18-hour acclimation periods, researchers can conduct standardized observation sessions of at least 5 minutes duration to assess mirror biting frequency, approach latency, and territorial display behaviors. The trapezoidal design optimizes visual access while maintaining appropriate swimming space for natural behavioral expression.
How It Works
The Mirror Biting Test exploits the natural territorial and social behaviors of fish species, particularly zebrafish, which respond to mirror reflections as conspecific intruders. When a fish encounters its reflection, it interprets the mirrored image as another individual within its territory, triggering species-typical aggressive and territorial responses including biting, charging, and lateral displays. The apparatus design ensures consistent visual presentation of the mirror stimulus while providing adequate swimming space for natural behavioral expression.
Behavioral quantification relies on defined spatial zones relative to the mirror surface. The contact zone (within 0.5 cm of the mirror) captures direct aggressive interactions including biting attempts, while the approach zone (within 3 cm) measures proximity-seeking and territorial positioning behaviors. Researchers can manipulate experimental conditions by introducing the mirror mid-trial to assess novelty responses or maintaining continuous mirror exposure to study habituation patterns.
The trapezoidal tank geometry optimizes visual access for behavioral recording while minimizing spatial constraints that might confound natural swimming patterns. Water temperature maintenance at ambient levels ensures physiological stability throughout testing sessions, while the standardized observation duration of at least 5 minutes provides sufficient sampling of behavioral repertoires for statistical analysis.
Features & Benefits
1.5-liter trapezoidal tank design
Provides optimal swimming space while ensuring consistent mirror presentation angles and minimizing spatial constraints on natural behaviors.
Integrated 15 x 7 cm mirror
Delivers standardized visual stimulus size for consistent conspecific simulation across experimental sessions and subjects.
Defined contact zone (0.5 cm from mirror)
Enables precise quantification of direct aggressive interactions and mirror biting attempts for detailed behavioral analysis.
Established approach zone (3 cm from mirror)
Allows measurement of proximity-seeking behaviors and territorial positioning responses separate from direct contact events.
Standardized observation duration protocols
Ensures adequate sampling of behavioral repertoires through minimum 5-minute sessions for statistically reliable measurements.
18-hour minimum acclimation requirement
Reduces stress-induced behavioral artifacts and establishes baseline swimming patterns before mirror introduction.
Ambient temperature maintenance
Maintains physiological stability throughout testing sessions without requiring specialized temperature control systems.
Flexible mirror introduction protocols
Supports both pre-placed mirror studies and mid-trial novelty introduction experiments for diverse research applications.
Sizes by model
| Measurement | Zebrafish |
|---|---|
| Height | 15 cm |
| Width | 7 cm |
| Base tank length | 23 cm |
| The tank length | 28 cm |
- 1.5 L trapezoidal tank
- the mouth of
- Mirror: 15 x 7 cm
Tank Shape
- Trapezoidal
Mirror Dimensions
- 15 x 7 cm
Contact Zone
- 0.5 cm from mirror
Approach Zone
- 3 cm from mirror
Acclimation Time
- minimum 18 hours
Observation Duration
- at least 5 minutes
Water Temperature
- ambient levels
Behavioral Construct
- aggression
- boldness
- social interaction
- territoriality
- habituation
Automation Level
- manual
Capacity
- 1.5 liters
Species
- Fish
- Zebrafish
Display Type
- None
Dimensions
- 23 cm (base) to 28 cm (mouth) x 7 cm x 15 cm
Research Domain
- Anxiety and Depression
- Behavioral Pharmacology
- Developmental Biology
- Learning and Memory
- Neuroscience
- Social Behavior
Shipping weight
- 21.0 lb
Shipping box
- L: 43.2 cm
- W: 38.0 cm
- H: 27.9 cm
| Feature | This Product | Typical Alternative | Advantage |
|---|---|---|---|
| Tank Capacity | 1.5 liters | Smaller systems often provide 0.5-1.0 L capacity | Larger volume allows more natural swimming behaviors while maintaining controlled mirror interaction distances. |
| Tank Geometry | Trapezoidal design | Most systems use rectangular or circular configurations | Trapezoidal shape optimizes visual recording angles while providing adequate swimming space. |
| Mirror Integration | Fixed 15 x 7 cm integrated mirror | Many systems use removable or adjustable mirrors | Fixed integration ensures consistent stimulus presentation and eliminates positioning variability between trials. |
| Behavioral Zones | Defined 0.5 cm contact and 3 cm approach zones | Zone definitions vary or may be undefined | Standardized zones enable precise quantification and comparison across studies and laboratories. |
| Acclimation Protocol | Minimum 18-hour acclimation requirement | Acclimation periods often shorter or unspecified | Extended acclimation reduces stress artifacts and establishes reliable baseline behavioral patterns. |
| Observation Duration | Minimum 5-minute sessions | Session lengths vary widely across protocols | Standardized duration ensures adequate behavioral sampling for statistical reliability across different research contexts. |
The Mirror Biting Test Apparatus provides a standardized platform for fish behavioral assessment with defined spatial zones, consistent mirror presentation, and established protocols. The trapezoidal design optimizes both behavioral expression and video recording capabilities for comprehensive social behavior quantification.
Neuroscience
Assessing social behavior phenotypes in zebrafish models of neuropsychiatric disorders such as schizophrenia and autism spectrum disorders through quantification of mirror-directed aggressive behaviors.
Behavioral Pharmacology
Evaluating pharmacological effects on territorial aggression and social interaction patterns in fish models by measuring changes in mirror biting frequency and approach behaviors.
Developmental Biology
Characterizing developmental changes in social behavioral repertoires by comparing mirror biting responses across different life stages in zebrafish populations.
Social Behavior
Investigating individual differences in boldness and aggression phenotypes through standardized mirror confrontation assays in laboratory fish populations.
Anxiety and Depression
Measuring social withdrawal and altered aggression patterns in fish models of mood disorders through reduced mirror interaction and modified territorial responses.
Learning and Memory
Studying habituation and recognition memory processes by assessing changes in mirror biting responses across repeated exposure trials.
Practical Tips
Best Practices
Allow complete acclimation before mirror introduction to establish baseline swimming patterns and reduce stress-induced behavioral artifacts.
Why: Proper acclimation ensures behavioral responses reflect mirror stimulation rather than handling or environmental stress.
Calibration
Clearly mark contact and approach zone boundaries using the mirror surface as reference before beginning behavioral scoring.
Why: Precise zone calibration ensures accurate behavioral quantification and enables comparison across sessions and subjects.
Maintenance
Clean mirror surface between subjects using appropriate aquarium-safe cleaning solutions to maintain consistent visual clarity.
Why: Mirror cleanliness affects stimulus intensity and behavioral response consistency across experimental sessions.
Data Quality
Record sessions from multiple camera angles to capture both lateral displays and direct mirror interactions for comprehensive behavioral analysis.
Why: Multiple viewing angles prevent data loss from behavioral events occurring outside primary camera field of view.
Troubleshooting
If subjects show no mirror interest, verify water clarity, lighting conditions, and mirror positioning relative to swimming areas.
Why: Environmental factors can mask mirror stimulus or reduce behavioral responsiveness leading to false negative results.
Safety
Monitor subjects continuously during testing sessions to detect signs of excessive stress or abnormal behaviors requiring session termination.
Why: Early detection of stress responses protects subject welfare and prevents confounding of behavioral measurements.
Best Practices
Maintain consistent ambient lighting conditions across all testing sessions to ensure standardized mirror visibility and behavioral responses.
Why: Lighting variations can alter mirror stimulus intensity and introduce confounding variables in behavioral comparisons.
Setup Guide
Tank Preparation
Fill the trapezoidal tank with dechlorinated water to appropriate depth, maintaining ambient temperature levels for species-specific requirements.
Mirror Installation
Securely position the 15 x 7 cm mirror within the tank, ensuring stable placement and clear visual access from the swimming area.
Zone Calibration
Mark or calibrate the contact zone (0.5 cm from mirror) and approach zone (3 cm from mirror) boundaries for accurate behavioral scoring.
Subject Acclimation
Allow test subjects minimum 18 hours acclimation period in the apparatus before behavioral testing to reduce handling stress effects.
Recording Setup
Position video recording equipment to capture clear views of fish movements and mirror interactions for subsequent behavioral analysis.
Testing Protocol
Conduct observation sessions of at least 5 minutes duration, recording mirror biting frequency, approach latency, and zone occupancy times.
Data Collection
Score behaviors in real-time or from video recordings, quantifying contact attempts, approach duration, and territorial display frequencies.
What’s in the Box
- Trapezoidal testing tank (1.5 L capacity)
- Integrated mirror (15 x 7 cm)
- Setup and protocol guidelines
- Behavioral scoring sheets (typical)
- Zone marking templates (typical)
Warranty
ConductScience provides a standard one-year manufacturer warranty covering defects in materials and workmanship, with technical support for setup and protocol optimization.
Compliance
ARRIVE GuidelinesSupports standardized reporting of animal research through defined protocols for fish behavioral studies and experimental design.
NIH Guide for Care and Use of Laboratory AnimalsCommonly used in research facilities subject to animal welfare guidelines for aquatic species behavioral testing.
IACUC Protocol RequirementsFacilitates institutional animal care committee approval through standardized behavioral testing procedures and defined endpoints.
Protocol and background
Introduction
The Mirror Biting test is used to assess social behavior phenotypes in fishes. Neuropsychiatric and Neuropsychological diseases such as Schizophrenia, personality disorders, and autism, all show some form of social deficiency. The Mirror Biting test employs mirror image simulation to quantify social interactions such as boldness, aggression, and sociability. Mirror tests are believed to provide better control of behavior variability and standardization of behaviors to allow repeated trials using the same subject.
Zebrafish model is a popular choice in mirror tests. Not only do they share significant genetic similarity to humans but are also known to be highly social creatures. Further, their robust social behaviors, such as shoaling, boldness, aggression, and social preference make them ideal for studies investigating social behaviors. Zebrafishes usually exhibit boldness by biting the mirror placed in the tank. They can also be seen to trace their reflection by quickly swimming back and forth. These behavioral responses are helpful in examining and evaluating social interactions and behaviors.
Pham’s mirror biting apparatus is composed of a trapezoidal tank and a mirror that snuggly fits the tank. The Mirror Biting task can be performed either by having the mirror already in the tank or introducing the mirror mid-trial to create a sense of novelty. In either case, the Mirror Biting task serves as an effective tool in biobehavioral research. The apparatus can also be used for the Novel Tank Diving test to evaluate anxiety. Other mirror based test include the Mirror Biting (Cattelan), the Mirror Biting (Balzarini) and Mirror Biting (Elwood).
Apparatus and Equipment
The Mirror Bite apparatus consists of a 1.5 L trapezoidal tank of height 15 cm and width 7 cm, and a 15 x 7 cm mirror. The base of the tank has a length of 23 cm while the mouth of the tank is 28 cm long. The tank is often virtually divided into an approach zone to assist with data recording and observations.
Training Protocol
Prior to the experiments clean the apparatus thoroughly. Fill the tank with clean water. Change the stimulus shoal and the water of the experimental tank once every three trials. Appropriately illuminate tank using overhead lighting. For tracking and recording, video and software such as the Noldus Ethovison XT can be used. Ensure that the water is at ambient temperatures.
Shoaling Test
Pretraining
Place groups of fishes from the same holding tank into a pretreatment beaker for at least 5 to 20 minutes. During this period, drugs to investigated can be mixed in with the water of the pretreatment tank before introducing the fishes.
Shoaling test
Place the group into the novel tank (without the mirror) and allow them about 3 minutes to acclimate to the environment. Observe the social interactions between the fishes for at least 3 minutes.
Mirror Biting Test
The Mirror Biting test can be conducted using two protocol that both provide an effective way of evaluating the social behaviors.
Protocol I: Introducing mirror to the tank
Place the subject in the tank and leave it undisturbed for at least 18 hours. Introduce the mirror into the tank without causing significant disturbances to the environment or the fish. Observe the interactions between the fish and the mirror for at least 5 minutes.
This protocol evokes stronger territorial behavior in the fish based on higher mirror novelty.
Protocol II: Introducing the subject to the mirrored tank
Place the mirror on the inner side of the tank and divide the tank into virtual spaces to denote the contacting area (0.5 cm from the mirror) and approach area (3 cm from the mirror). Place the fish in the tank and immediately begin observation of social behaviors.
This protocol uses stronger novelty of both the mirror and the tank to observe social behaviors. However, this protocol may not be suitable for anxious fishes.
Evaluation of the effect of acute ibogaine on zebrafish
Experimentally naïve zebrafish were used in the evaluation of the effects of acute ibogaine on mirror stimulus social interactions. Subjects were pre-exposed in 1-liter beaker to either drug-treated (10 and 20mg/L of ibogaine) or drug-free vehicle solution for 20 minutes. The subjects were then tested in the novel tank without a mirror to evaluate anxiety and locomotion. The subjects were also evaluated for aggressive behaviors using the same tank but with a mirror fixed inside. Both these tests lasted 6-minutes each. In the novel tank test, at 20 mg/L ibogaine reduced freezing duration and at 10 ml/L increased freezing frequency. Both doses also reduced latency to enter the top half of the tank significantly in comparison to controls. In the mirror test, a significant effect of the drug could be observed on the total distance traveled, average velocity, bottom contacts, total approaches, and total mirror contacts. (Cachat et al., 2013)
Evaluation of difference in mirror image fighting versus fighting a real opponent
Most animals do not have the ability of self-recognition. African cichlid fish, Astatotilapia, were used to investigate the differences in behavioral, hormonal and brain activity when the fishes fight a real opponent versus a mirror image. The dominant males were divided into three groups that either encountered a mirror, an opponent or an empty tank (control group). The fish were introduced into the tank with a barrier dividing the tank. The barrier was removed to reveal one of the 3 conditions. The subjects were scored for total aggression. No difference in total aggression or its associated components could be observed between the real opponent and mirror image opponent. However, a significant difference in the effects on the brain could be observed in the two fighting scenarios. Fishes that fought mirror images showed higher levels of egr-1 expression in the dorsolateral telencephalon while fishes with real opponents had a significantly higher c-fos expression in the hippocampus homolog. Further, a higher egr-1 and c-fos expression in the dorsomedial telencephalon of mirror fighting fish could be observed. (Desjardins and Fernald, 2010)
Investigation of performances of different strains of zebrafish in the mirror test
Three strains of zebrafish; TM1, Nadia and SH strains, were used to investigate strain differences in social behaviors. Following boldness tests, the subjects were visually isolated for at least 18-hours. The mirror biting test was performed by introducing the mirror into the tank. It was observed that the TM1 strain showed higher mirror biting frequency in comparison to the other two strains. (Mortez et al., 2005)
Data Analysis
The following parameters can be recorded during the Shoaling test.
- Average inter-fish distance: Distance between the body center of every member of the shoal.
- Average neighbor distance: Distance for the body center of each fish to the closest neighboring fish.
- Median distance between any two fish: Middle distance of all the inter-fish distances.
- Average farthest neighbor: Distance for the body center of each fish to the farthest neighboring fish.
- Top dwelling: Percent of fish in the top (upper half) of the tank.
- Thigmotaxis: The average distance of the group from the center of the tank.
- Variance of inter-fish distance: An index reflecting how homogeneously the fish are distributed in the shoal.
- Shoal area: The size of the shoal (width and length).
- Excursions from shoal: Number of excursions of individual fish away from the shoal.
- Duration of excursions: Duration of excursions of individual fish away from the shoal.
- Shoal polarization: Absolute size of the summed vector of all fish in the shoal.
- Spread of the group: Smallest wedge (extending to edges) that captures within it all the fish.
The following behaviors and parameters can be observed in the Mirror Biting test.
- Mirror biting frequency: Number of times the fish bite the mirror.
- Mirror biting duration: Time spent biting mirror.
- Approaches to the mirror: The number of crossing the line denoting the mirror approach zone, but without mirror contact.
- Mirror contacts: The number of crossing the line denoting the mirror contact zone.
- Latency to approach: Time to the first approach to the mirror.
- Aggressive tail beats: The number of aggressive tail beats against the mirror.
- Latency to contact: Time to the first contact with the mirror.
Strengths and Limitations
Strengths
Mirror tests have been shown to be a reliable measure of how fish behave with social companions and their social behaviors have been positively correlated with the shoaling tendency. The mirror in the tank can be removed or added as per need. The two protocols that can be used for the Mirror Biting test allow the introduction of different levels of novelty. Introducing the mirror to the tank already holding the fish evokes stronger territorial behavior. The apparatus is simple to construct and allows modifications to suit the needs of investigations. The narrow tank allows creating different densities of shoals to evaluate shoaling behaviors.
Limitations
Although a correlation exists, the mirror test is still a crude method for understanding sociability as some mirror-directed behavior could be the result of fear avoidance. Further, the mirror test does not emulate the characteristics of a real stimulus conspecific thus requiring caution when interpretations are made. The subject may also not be able to see the mirror image from close proximity clearly. The test also deprives the subject of chemical cues involved in typical social interactions between fish when the subject is tested using the mirror image stimulus. Subject’s and the stimulus conspecific’s social personality type also greatly influences the results. Anxious fishes may not perform as expected when introduced to the tank already housing the mirror.
Summary and Key Points
- Mirror Biting test is used in the investigation of social behaviors and phenotypes in fish.
- Mirror Biting test is believed to allow better control of the variability of behavior and emulation of standardized behavior.
- The Mirror Biting apparatus consists of a trapezoidal tank that has a mirror that can be easily removed or added.
- The Mirror Biting test can be conducted using either of the two protocols, introducing mirror into the tank or introducing the fish to the mirrored tank.
- The two protocols provide different levels of novelty during the testing.
- Mirror Biting test doesn’t emulate all the characteristics of real social interactions.
- The subject’s own preference and its mental state can influence the performance in the test.
References
Cachat J, Kyzar EJ, Collins C, Gaikwad S, Green J, Roth A, El-Ounsi M, Davis A, Pham M, Landsman S, Stewart AM, Kalueff AV (2013). Unique and potent effects of acute ibogaine on zebrafish: the developing utility of novel aquatic models for hallucinogenic drug research. Behav Brain Res. 236(1):258-69. doi: 10.1016/j.bbr.2012.08.041.
Desjardins JK, Fernald RD (2010) What do fish make of mirror images? Biology Letter 6(6): 744–747
Marks C, West TN, Bagatto B and Moore FBG (2005). Developmental environment alters conditional aggression in zebrafish. Copeia 4:901–908
Moretz AA, Martins EP, Robinson BD (2007). Behavioral syndromes and the evolution of correlated behavior in zebrafish. Behav Ecol 3:556–562
Oliveira RF, Silva JF, Simoes JM (2011) Fighting zebrafish: characterization of aggressive behavior and winner-loser effects. Zebrafish 8(2):73–81
Pham M. et al. (2012) Assessing Social Behavior Phenotypes in Adult Zebrafish: Shoaling, Social Preference, and Mirror Biting Tests. In: Kalueff A., Stewart A. (eds) Zebrafish Protocols for Neurobehavioral Research. Neuromethods, vol 66. Humana Press, Totowa, NJ
From the Maze Engineers documentation for this apparatus.
Q
What fish species can be tested with this apparatus besides zebrafish?
A
The apparatus is designed for fish species that exhibit territorial and mirror-directed behaviors, though zebrafish remain the most validated model due to their robust social behavioral repertoires and genetic tractability.
Q
How should behavioral zones be validated for accurate scoring?
A
The contact zone (0.5 cm) and approach zone (3 cm) should be clearly marked and calibrated using the mirror surface as reference, with video recording systems positioned to capture zone boundaries for reliable behavioral quantification.
Q
What is the recommended acclimation protocol before testing?
A
Allow minimum 18 hours acclimation in the apparatus to reduce handling stress and establish baseline swimming patterns, with water maintained at ambient temperature levels appropriate for the test species.
Q
How long should observation sessions last for reliable data?
A
Observation sessions should last at least 5 minutes to capture adequate sampling of behavioral repertoires, though longer sessions may be required depending on species-specific activity patterns and research objectives.
Q
Can the mirror be introduced during trials rather than pre-placement?
A
Yes, the apparatus supports both continuous mirror exposure and mid-trial introduction protocols, allowing researchers to study novelty responses and habituation patterns depending on experimental design requirements.
Q
What behavioral parameters should be quantified during testing?
A
Key measures include mirror biting frequency, approach latency, time spent in contact and approach zones, territorial display duration, and overall activity levels for comprehensive behavioral phenotyping.
Q
How does tank geometry affect behavioral expression?
A
The trapezoidal design provides optimal visual access while minimizing spatial constraints, ensuring adequate swimming space for natural behavioral patterns while maintaining consistent mirror presentation angles.
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