To begin, monitor the sleep behaviors of flies using an infrared monitoring device and import the collected data in TXT format into the DAM file scan 107 software for scanning. Set the start time of the segmentation data to either 0800 or 0801 on the third morning after initiating the monitors to obtain one or 30 minute bin length data, respectively. Set the termination time to 8:00 AM on the third day post the set starting time.
Adjust the bin length option to one minute and alter the output file type to channel files. Rename the file and generate the output. Now launch the SCAMP 2020 software package in MATLAB.
Double click on Vecsey Sleep and Circadian Analysis MATLAB program. Added it sub-folder titled Vecsey SCAMP Scripts into the path. Locate the SCAMP.
m file in that folder and run it. In the subsequent popup window sequentially select the folder named one minute and 30 minute. Then select a monitor and click on load individual C plots to preview.
Inspect the image that appears and deselect the corresponding channel of any dead flies. Rename each channel in every monitor based on the corresponding drug to be tested. Select all monitors and click Analyze selected data for the analysis.
Select the default option and then click on analyze for chosen bin feature. Check the export data box. Click on graph 30 minute data types for all days for selected groups, and export all data to generate the output of the results.
Select the S30 file from the CSV file and locate the corresponding mean value and standard error data for each monitor. Save this data in Excel to make some adjustments before posting it into GraphPad Prism to formulate a sleep status diagram. Afterwards, locate the file named S-T-D-U-R and compute the average values of daytime, nighttime, and total sleep for each fly over three days.
Paste data into GraphPad Prism to draw graphs. The sleep patterns of Reserpine and Dimethyl Sulfoxide on female and male flies over three consecutive days are shown. Quantitative analysis of the sleep behavior demonstrates a significant increase in sleep time in aged females fed Reserpine.
The experiment on male flies also demonstrates a positive correlation between Reserpine concentration and the promotion of sleep.
