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Limelight software

Manufactured by Actimetrics
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Limelight is a software application developed by Actimetrics for data analysis and visualization. It provides tools for processing and examining data from various sources, with a focus on presenting information in a clear and intuitive manner.

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Lab products found in correlation

Ethovison xt software, by Noldus (2 mentions) Open field arena, by Harvard Apparatus (2 mentions) Ethovision xt, by Noldus (2 mentions) Activity chamber, by Med Associates (1 mentions)

30 protocols using limelight software

1

Cocaine-Induced Behavioral Sensitization in Mice

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For cocaine sensitization experiments, mice were first habituated to an open arena for three days (5 minutes per day) to ensure that there was no baseline difference in locomotor activity. Then, mice were injected with either saline or 20mg/kg of cocaine daily for 5 days. Cocaine or saline was injected 5 minutes before being placed in an open field (40 cm x 40 cm). Changes in locomotor activity in response to cocaine was used as a measure of behavioural sensitization and was recorded over a period of 5 minutes using an overhead camera connected to an automated tracking system (Limelight software, Actimetrics). Dim lighting (40 lux) was used during all sessions to promote exploration of the open field. One week after the end of the sensitization the mice were tested in the same open field with locomotor activity recorded. All mice were first tested in a drug free state followed by a 20mg/kg cocaine challenge 24 hours later.
Cromar G.L., Epp J.R., Popovic A., Gu Y., Ha V., Walters B.J., St. Pierre J., Xiong X., Howland J.G., Josselyn S.A., Frankland P.W, & Parkinson J. (2022). Toxoplasma infection in male mice alters dopamine-sensitive behaviors and host gene expression patterns associated with neuropsychiatric disease. PLoS Neglected Tropical Diseases, 16(7), e0010600.
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2

Open Field Locomotor Evaluation

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Assessment of mouse behavior in an open field was performed as previously described (27 (link)). Individual mice (n = 9 for each genotype) were placed at the center of a white acrylic chamber (71 × 71 × 30 cm) lit by indirect white light (200 lux at center of chamber) and allowed to explore for 5 min. The open field was divided into an 8 × 8 grid with a center zone (53.25 × 53.25 cm) and a peripheral zone (the outer 8.875 cm on all sides). Total distance traveled in the center zone was measured by video signals from digital cameras sent to a desktop PC and processed online with Actimetrics LIMELIGHT software. Experimenters were blind to genotype throughout.
Wagnon J.L., Korn M.J., Parent R., Tarpey T.A., Jones J.M., Hammer M.F., Murphy G.G., Parent J.M, & Meisler M.H. (2014). Convulsive seizures and SUDEP in a mouse model of SCN8A epileptic encephalopathy. Human Molecular Genetics, 24(2), 506-515.
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3

Assessing Mouse Anxiety and Locomotion

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The open field arena (56 × 56 cm) was used to assess levels of anxiety in the mouse as well as ambulation and activity levels as well as exploratory habits. Animals were placed in the center of the arena inside a sound proof chamber. Each sound proof chamber was evenly illuminated and equipped with a ceiling-mounted camera connected to a computer to monitor and capture the response of the mouse. LimeLight software (Actimetrics, Wilmette, IL) was used to collect ambulation activity for 300 s while the mouse freely traversed the arena. Data were calculated as the total distance traveled as well as the number of crossings in the arena as defined by the software. A hyperactive mouse will explore the test chamber and travel greater distances while an anxious mouse will spend more of its time along the perimeter and will not cross the arena.
Makinde H.M., Winter D.R., Procissi D., Mike E.V., Stock A.D., Kando M.J., Gadhvi G.T., Droho S., Bloomfield C.L., Dominguez S.T., Mayr M.G., Lavine J.A., Putterman C, & Cuda C.M. (2020). A Novel Microglia-Specific Transcriptional Signature Correlates With Behavioral Deficits in Neuropsychiatric Lupus. Frontiers in Immunology, 11, 230.
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4

Open Field Exploration in Mice

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Mice were placed in the center of an open arena (56 × 56cm), and ambulation activity for 5 min was recorded by LimeLight software (Actimetrics, Wilmette, IL). The open field was divided into a 5 × 5 grid of squares, with the outer 16 squares defining the ‘outer’ region and the inner 9 squares defining the ‘middle’ region. The software provided the total distance traveled, the percentage of time the mouse spent within each of the two regions, and the number of crossings from one region to the other.
Maneshi M.M., Toth A.B., Ishii T., Hori K., Tsujikawa S., Shum A.K., Shrestha N., Yamashita M., Miller R.J., Radulovic J., Swanson G.T, & Prakriya M. (2020). Orai1 Channels Are Essential for Amplification of Glutamate-Evoked Ca2+ Signals in Dendritic Spines to Regulate Working and Associative Memory. Cell reports, 33(9), 108464.
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5

Open Field Exploration and Emotionality Assessment in Rats

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The open field was a clear plexiglass open-field (28 × 28 × 46 cm) with a black plexiglass barrier present between the experimenter and the apparatus. Illumination was ~700 lux. The rats were placed in the rear left corner of the open field and allowed to explore the box for 20 min. Movements were recorded using Limelight Software (Actimetrics, Wilmette, IL). The software divided the open field arena into 16 squares of 7 × 7 cm. The central four squares were defined as the center zone, in which animals’ activity was regarded as a measure of emotionality (Prut and Belzung, 2003 (link)). The software recorded (a) total number of crosses across squares, (b) total distance traveled (cm), (c) crosses into the center zone, and (d) time spent in the center zone (sec) (n=42-46). Between each subject, the chambers were thoroughly cleaned and wiped down with 70% ethanol. Due to a computer malfunction, the open field data for 16 animals was not collected and unavailable for inclusion in the statistical analysis.
Carr R.L., Alugubelly N., de Leon K., Loyant L., Mohammed A.N., Patterson M.E., Ross M.K, & Rowbotham N.E. (2020). Inhibition of Fatty Acid Amide Hydrolase by Chlorpyrifos in Juvenile Rats Results in Altered Exploratory and Social Behavior as Adolescents. Neurotoxicology, 77, 127-136.
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6

Elevated Zero Maze for Mouse Anxiety

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The EZM consisted of a 56 cm diameter round track divided into two closed quadrants separated by 180 degrees and two open quadrants separated by 180 degrees. The closed quadrants were enclosed by 15 cm high walls and the open quadrants had no walls. The maze was elevated 46 cm above the ground and testing was conducted under white light. The mouse was placed into a closed quadrant and behavior was video recorded for 5 minutes. LimeLight software (Actimetrics) measured the percentage of time the mouse spent in open quadrants.
Brooker S.M., Gobeske K.T., Chen J., Peng C.Y, & Kessler J.A. (2016). Hippocampal bone morphogenetic protein signaling mediates behavioral effects of antidepressant treatment. Molecular psychiatry, 22(6), 910-919.
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7

Assessing Anxiety in Mice Using Zero Maze

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Anxiety was assessed using the zero maze test. The zero maze was a round track (56 cm diameter) divided into four equal (two open and two closed) sections placed on an elevated stand and this task is based on the preference of mice for enclosed spaces. The closed sections are made up of two sets of walls along the track. The mice were placed on the track in the center of the open area and were examined for a preference for the closed or open arms. Each sound proof chamber was evenly illuminated and equipped with a ceiling-mounted camera connected to a computer to monitor and capture the response of the mouse. LimeLight software (Actimetrics, Wilmette, IL) was used to collect data for 300 s. Data were calculated as the percent of time in the open portions of the track. While an anxious mouse will avoid the open regions of the track, animals with a reduction in anxiety levels due to a neurological defect will spend an increased amount of time in the open space of the maze.
Makinde H.M., Winter D.R., Procissi D., Mike E.V., Stock A.D., Kando M.J., Gadhvi G.T., Droho S., Bloomfield C.L., Dominguez S.T., Mayr M.G., Lavine J.A., Putterman C, & Cuda C.M. (2020). A Novel Microglia-Specific Transcriptional Signature Correlates With Behavioral Deficits in Neuropsychiatric Lupus. Frontiers in Immunology, 11, 230.
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8

Open Field Exploration in Mice

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Mice were placed in the center of an open arena (56 × 56cm), and ambulation activity for 5 min was recorded by LimeLight software (Actimetrics, Wilmette, IL). The open field was divided into a 5 × 5 grid of squares, with the outer 16 squares defining the ‘outer’ region and the inner 9 squares defining the ‘middle’ region. The software provided the total distance traveled, the percentage of time the mouse spent within each of the two regions, and the number of crossings from one region to the other.
Maneshi M.M., Toth A.B., Ishii T., Hori K., Tsujikawa S., Shum A.K., Shrestha N., Yamashita M., Miller R.J., Radulovic J., Swanson G.T, & Prakriya M. (2020). Orai1 Channels Are Essential for Amplification of Glutamate-Evoked Ca2+ Signals in Dendritic Spines to Regulate Working and Associative Memory. Cell reports, 33(9), 108464.
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9

Novel Object Recognition Test

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The NOR test (Carpenter et al., 2021 ) was performed on day 15. Briefly, the previous day's OF test served as the NOR habituation phase. During the identical phase of the NOR, mice were placed in the open field with two identical objects and allowed to freely explore them for 5 min. Mice were then returned to their home cage for 1 h. During the novel phase, one identical object was replaced with a novel object. Mice were placed in the open field and allowed to explore both objects. The number of novel (n) or familiar (f) object approaches (N) and time spent (T) were scored for the first 20 s and the total 5 min of the novel phase with the Limelight software (Actimetrics) as in Carpenter et al. (2021) . The novelty preference index (NPI) was calculated as described in Lin et al. (2013) (link) using the following equations: NPI = (Nn − Nf)/(Nn + Nf) or NPI = (Tn − Tf)/(Tn + Tf).
Carpenter J.M., Brown K.A., Veltmaat L., Ludwig H.D., Clay K.B., Norberg T., Harn D.A., Wagner J.J, & Filipov N.M. (2022). Evaluation of delayed LNFPIII treatment initiation protocol on improving long-term behavioral and neuroinflammatory pathology in a mouse model of Gulf War Illness. Brain, Behavior, & Immunity - Health, 26, 100553.
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10

Elevated Zero Maze Anxiety Assessment

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Mice were evaluated for anxiety-related behavior in an elevated zero maze. The maze consists of an annular platform (diameter 46 cm; elevation 50 cm) divided into equally sized quadrants, alternating between open and enclosed (wall height 17 cm). This configuration lacks the ambiguous center region associated with the elevated plus maze42 (link). The open and closed arms were illuminated to similar levels (30 lx). Individual mice were placed near an enclosed arm of the maze and allowed to freely explore for 5 min. Limelight software (Actimetrics, Wilmette, IL, USA) was used to video record each trial. Ethovison XT software (Noldus, Leesberg, VA, USA) was used to track the position of the mouse, and calculate distance traveled, mean velocity and time spent in closed or open arms (n = 6–14 per strain, genotype, and sex). Trials where mice exited the maze were excluded from analysis. Exclusions by group were as follows: B6.Scn2aE/+ males – 7 of 13, B6.Scn2aE/+ females – 5 of 15, F1D2.Scn2aE/+ males – 2 of 11; all other groups had no exclusions. The proportion of B6.Scn2aE/+ males excluded due to maze exit was significantly greater than B6.WT males (p=0.0052, Fisher’s exact test), while the proportion of B6.Scn2aE/+ females excluded was significantly greater than F1D2.Scn2aE/+ females (p=0.0421, Fisher’s exact test).
Echevarria-Cooper D.M., Hawkins N.A, & Kearney J.A. (2023). Strain-dependent effects on neurobehavioral and seizure phenotypes in Scn2aK1422E mice. bioRxiv.
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