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UltraSoundGate 116Hb

Manufactured by Avisoft
Sourced in Germany

The UltraSoundGate 116Hb is a high-performance data acquisition device designed for recording and analyzing ultrasonic signals. It provides 16 input channels and supports a sampling rate of up to 500 kHz per channel. The device is capable of capturing and storing ultrasonic data for further processing and analysis.

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33 protocols using UltraSoundGate 116Hb

Multiple systems are capable of detecting and measuring ultrasonic sound. We selected two commercially available systems to identify sources of ultrasonic noise at the Mary Lyon Centre: 1) The Avisoft system - an Avisoft condenser microphone CM16/CMPA connected to an Avisoft UltraSoundGate 116Hb (Avisoft Bioacoustics, Berlin), and 2) the HBK PULSE system with a type 4939 ¼” measuring microphone attached to a high frequency 3110 processing module (Hottinger, Bruel & Kjaer UK).
These systems were selected as both can detect ultrasound and are of a very high quality. The Avisoft system is designed to monitor ultrasonic vocalisations (USVs) used in social communication and the PULSE system is a sound analysis platform designed for accurate noise and vibration measurement and analysis. Although the objective of both systems is to detect audible and ultrasonic noise, the intended purposes result in distinct differences between the two systems. The microphones differ in sensitivity: the Avisoft microphone CM16/CMPA (500 mV/Pa) is orders of magnitude higher than the HBK 4939 (4 mV/Pa), but by design, the HBK 4939 microphone has a much flatter frequency response, allowing single frequency calibration for accurate measurement. An AudioMoth (Open Acoustic Devices) full spectrum recorder was also used for one of the noise sources.
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On P2 a dam’s litter was removed from the Phenotyper cage and placed in a standard home cage. Ultrasonic vocalizations (USVs) made by the dam for was recorded 180 s using Avisoft-UltraSoundGate 116Hb (Avisoft Bioacoustics e.K., Germany). The dam was then removed from the Phenotyper cage and placed in a standard home cage separate to pups. Where possible, two male and two female pups, determined visually by anogenital distance, were placed in Phenotyper cage opposite the nest and spaced apart, alternating male and female. USVs made by these four pups were simultaneously recorded for 180 s as above. Following this video recording was initiated and dam was reintroduced to the Phenotyper cage, indicating time zero. Time taken to sniff and retrieve each pup was recorded by experimenter and confirmed off line from video recordings. When all pups had been retrieved by the dam, remaining pups were returned to the Phenotyper.
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USV was performed on fully WT pups and fully Phlda2−/− pups on P2 in a separate cohort generated by natural mating. The dam’s litter was removed from the home cage and placed in a separate home cage. USVs made by the pups were recorded for 180 seconds using Avisoft-UltraSoundGate 116Hb (Avisoft Bioacoustics e.K., Germany).
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USVs were recorded using an ultrasonic microphone (Avisoft UltraSoundGate 116Hb, Avisoft Bioacoustics, Berlin, Germany) and accompanying software (Avisoft Bioacoustics USGH). As in previous research, recording settings included a sampling rate of 250 kHz, format 16 bit, and a high pass filter set to 15 kHz in order to reduce background noise (McNamara et al., 2018 (link)). Raven Pro 1.6 software (Bioacoustics Research Program, Cornell Lab of Ornithology, Ithaca, NY) was used to generate spectrograms with a 256-sample Hann window, one frame per second. Raven’s built in call detection was then used to detect calls, with a selection of spectrograms manually checked to determine accurate detection.
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Vocalization recordings were done using a condenser microphone (CM16/CMPA, Avisoft Bioacoustics, Germany) connected to an ultrasound‐recording interface (UltraSoundGate 116Hb, Avisoft Bioacoustics). Calls counting was done using DeepSqueak (Coffey et al., 2019 (link)) on MATLAB 2018b. A short rat call network was used for the detection of USVs in the recordings with the detection settings set as “high recall” to ensure the detection of as many calls as possible. The other detection parameters were set as followed: total analysis length of 0, analysis chunk length of 6, frame overlap of 0.001 s, frequency low cut‐off of 30 kHz, frequency high cut‐off of 120 kHz, and score threshold of 0. After detection was finalized, the files were revised by an experimenter blind to testing conditions to exclude false‐positive findings. Vocalizations were manually classified as previously described (Romano et al., 2013 (link)). Calls were classified into a syllable repertoire of 11 different syllables: chevron, complex, composite, downward, flat, frequency step, short, two‐component, upward, unstructured, and calls that did not fit any of the syllables classifications were called unknown. Due to the low number of unknown calls, this type of USV was not included in the repertoire analysis.
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6

Isolation-Induced Ultrasonic Vocalizations in Seizure Pups

To assess changes in USV behavior following seizures, we examined USVs on PD12 using the isolation-induced USV paradigm previously described [15 (link), 16 (link)]. All vocalizations were recorded in the afternoon (between 1 and 5 pm), approximately 24 hours after the last seizure for the HSL paradigm and 48 hours later for the LSL paradigm. Briefly, all pups were weighed and transferred to a holding cage with fresh bedding, warmed by an electronic heating pad to ambient nesting temperature (~35° C). Ultrasonic vocalizations were recorded using a condenser microphone (CM16/CMPA, Avisoft Bioacoustics, Germany) connected to an ultrasound-recording interface (UltraSoundGate 116Hb, Avisoft Bioacoustics), which allowed assessment of all USVs on a continuous spectrum from 0–125 kilohertz (kHz). Each pup was individually placed into another housing pan, within an acrylic sound-attenuating chamber (40 cm × 40 cm × 30 cm) where USVs were recorded for 2 minutes. Following recording, the pups were placed back into the holding cage with their littermates. This procedure was repeated until each pup in the litter was tested. An experimenter remained in the room during all recordings. At the conclusion of testing, pups were returned to the home cage.
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The methods for adult USV collection have been previously described (Wöhr et al., 2011 ). Briefly, one week prior to testing, male mice were paired with females for a 5-minute duration. This was done to standardize the social experience in males, a necessary prerequisite for eliciting adult vocalizations (Scattoni et al., 2008 ). Twenty-four hours prior to testing, estrus was induced in females by placing male bedding into the female cage that had been previously introduced to the test males. On the day of testing, fresh urine was collected from females that were in estrus and 20 uL of the urine was then pipetted into the center of a testing chamber. The chamber was sound attenuating to ensure an accurate detection of the test mouse’s USVs. Males were placed in the chamber and all vocalizations were recorded for a 5-minute duration using a condenser microphone (CM16/CMPA, Avisoft Bioacoustics, Germany) coupled with a recording interface (UltraSoundGate 116Hb, Avisoft Bioacoustics). After testing, the males were returned to their home cages.
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P10 rat and mouse pups were placed into a recording cage in a sound-isolation box. The detection sessions were initiated directly thereafter and were always performed by the same experienced experimenter between 8 and 10 AM. A maximum of two pups per litter were tested for 5 min each. For P10 rats and mice signals were recorded at a 35–250 kHz range using an Avisoft Ultra Sound Gate 116Hb (Avisoft Bioacoustics). Incoming signals were displayed as a time–event plot by Avisoft recorder software. For further statistical analysis, the parameters number of events (n) and total duration of cues (s) were analyzed using one-way ANOVA.
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We examined ultrasonic vocalizations (USVs) on PD12 using an isolation-induced paradigm to assess changes in vocalization production following seizure and treatment administration on PD10. Prior to the recording phase, all pups were transferred to a new housing cage with fresh bedding that was warmed with a heating pad to an ambient temperature of ∼35°C. During recording, one at a time pups were placed into a separate housing pan placed within an acrylic sound-attenuating chamber. Vocalizations were recorded for 2 min for each pup. The recording apparatus consisted of a condenser microphone (CM16/CMPA, Avisoft Bioacoustics, Germany) which was connected to an ultrasound-recording interface (UltraSoundGate 116Hb, Avisoft Bioacoustics) and recorded all USVs on a continuous spectrum from 0 to 125kHz. Following recording, pups were placed back into the warmed housing pan with littermates. This procedure was repeated in sequence until all pups were recorded, and upon completion all pups were returned to their original home cage. The total number of recording sessions allowed for pups to not be separated from their home cage and mothers for longer than 30 minutes.
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Vocal emissions were recorded for the duration of the social behavior tests using an UltraSoundGate CM16/CMPA microphone (Avisoft Bioacoustics) placed just above the testing cage. The microphone was connected to a computer via an Avisoft Bioacoustics UltraSoundGate 116Hb. Acoustic data were recorded with a sampling rate of 250 kHz in 16 bit format, and spectrograms were constructed by fast Fourier transformation (FFT; 256 FFT length, 100% frame, FlatTop window, and 50% time window overlap; SASLab Pro, Avisoft Bioacoustics). All USVs made within the first 10 min of the play behavior trial were manually marked by investigators who were blind to the age, sex, and genotype of the rats. In order to be marked, calls had to be at least 10 ms in length, and distinct calls had to be separated by at least 10 ms. Several call parameters were quantified, including fundamental frequency, duration, and number of calls emitted. Call frequency (in hertz) was calculated by averaging the fundamental frequency at call onset, call offset, and peak amplitude of the call (integrated frequency). A subset (20% random sampling) of the calls was selected and manually classified into the 15 call categories described in the study by Wright et al. (2010) (link).
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