A Customizable Low-Cost System for Massively Parallel Zebrafish Behavioral Phenotyping

Abstract

High-throughput behavioral phenotyping is critical to genetic or chemical screening approaches. Zebrafish larvae are amenable to high-throughput behavioral screening because of their rapid development, small size, and conserved vertebrate brain architecture. Existing commercial behavioral phenotyping systems are expensive and not easily modified for new assays. Here, we describe a modular, highly adaptable, and low-cost system. Along with detailed assembly and operation instructions, we provide data acquisition software and a robust, parallel analysis pipeline. We validate our approach by analyzing stimulus response profiles in larval zebrafish, confirming prepulse inhibition phenotypes of two previously isolated mutants, and highlighting best practices for growing larvae prior to behavioral testing. Our new design thus allows rapid construction and streamlined operation of many large-scale behavioral setups with minimal resources and fabrication expertise, with broad applications to other aquatic organisms.

Scalable Apparatus to Measure Posture and Locomotion (SAMPL): a high-throughput solution to study unconstrained vertical behavior in small animals

“…We see SAMPL as a complementary tool. SAMPL’s emphasis on vertical behavior and its scalability position it to leverage the advantages of the zebrafish model for screens – either as a primary resource, or to follow-up on promising “hits” identified with higher-throughput approaches 6 .…”

“…The camera hardware satisfies the following demands: (1) large pixel size with low noise allowing for high dynamic range / signal-tonoise ratio; (2) sufficient resolution to resolve subtle changes to animal posture; (3) an interface with sufficient bandwidth for data transfer; (4) availability. The lens achieves (1) close focus; (2) sufficient depth-of-field to cover the entire depth of the imaging arena; (3) high image quality; (4) compact size; (5) high IR transmission rate; (6) ease of integrating an IR-pass filter. We adapted a 50 mm IR-optimized lens by placing a 0.3" extension tube between the lens and the camera to achieve higher magnification ratio with minimum working distance.…”

“…Behavioral screening is a fundamental part of both basic and translational approaches to disease 2 ‘ 3 . For screens, measuring behavior from large numbers of animals is necessary to differentiate individual variation 4 from changes seen in disease models and/or improvement following treatment 5,6 . The demand for such high-throughput measurements comes at a cost: often, measurements that require high resolution – such as posture – are limited.…”

Examining craniofacial variation among crispant and mutant zebrafish models of human skeletal diseases

“…Our results suggest that regardless of the type of fish used in a study (somatic vs germline), there is a strong clutch effect in zebrafish. Clutch effects have also been observed in zebrafish cardiac performance (Schwerte et al, 2005) and behavioral phenotypes (Joo et al, 2021). Further, the sost fish, which were from a different facility show the greatest separation in morphospace from the other groups in our combined analysis (Figure 1B-D).…”

Knockout of tanc2 causes autism‐like behavior and sleep disturbance in zebrafish

“…The chamber was continuously illuminated with infrared lights and was also illuminated with white lights from 8:00 A.M. to 10:00 P.M. Larval locomotor activity was monitored over from 4 dpf to 6 dpf. Data analysis was performed based on video recorded by EthoVision XT software (Noldus, Netherland) using previously described algorithms (Joo et al, 2020; Rihel et al, 2010). Any 1 min of inactivity was considered as sleep and a sleep bout was defined as a continuous string of sleep.…”