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1 Center for Sleep and Respiratory Neurobiology and Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, United States
2 Center for Sleep and Respiratory Neurobiology, University of Pennsylvania, Philadelphia, Pennsylvania, United States
3 Rutgers University, United States
4 The Jackson Laboratory, United States
5 The Jackson Laboratory, Bar Harbor, Maine, United States
* To whom correspondence should be addressed. E-mail: pack{at}mail.med.upenn.edu.
Assessment of sleep in mice currently requires initial implantation of chronic electrodes for assessment of electroencephalogram (EEG) and electromyogram (EMG) followed by time to recover from surgery. Hence, it is not ideal for high throughput screening. To address this deficiency, a method of assessment of sleep and wakefulness in mice has been developed based on assessment of activity/inactivity either by digital video analysis or by breaking infrared beams in the mouse cage. It is based on the algorithm that any episode of continuous inactivity of 
40 seconds is predicted to be sleep. The method gives excellent agreement in C57BL/6J male mice with simultaneous assessment of sleep by EEG/EMG recording. The average agreement over 8,640 10 second epochs in 24 hours is 92% (n=7 mice) with agreement in individual mice being 88-94%. Average EEG/EMG determined sleep per 2-hour interval across the day was 59.4 minutes. The estimated mean difference (bias) per 2-hour interval between inactivity defined sleep and EEG/EMG defined sleep was only 1.0 minute (95% CI for mean bias -0.06 to 2.6 minutes). The standard deviation of differences (precision) was 7.5 minutes per 2-hour interval with 95% limits of agreement ranging from -13.7 to +15.7 minutes. Although bias significantly varied by time of day (p=0.0007), the magnitude of time of day differences was not large (average bias during lights on and lights off were +5.0 and -3.0 minutes per 2-hour interval, respectively). This method has applications in chemical mutagenesis and for studies of molecular changes in brain with sleep/wakefulness.
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