Generation of sperm motility profiles using CASAnova
Track data generated for CASAnova must be derived from mouse sperm analyzed in chambers with a depth of 80-100 µm or human sperm analyzed in chambers 20 µm deep.
These instructions correspond to data generated using Hamilton-Thorne (HT) CASA systems. Compatibility with data from other systems is not guaranteed.
Input files:
Both mouse and human CASAnova were developed with Hamilton Thorne Animal and Human Motility software, (versions 12.3 and 14.0, respectively). During CASA analysis, data for every motile track analyzed is saved in an ASCII file. These files can be located and configured by following the HT user manual. Track data can contain many different parameters; the minimum parameters required for CASAnova are VAP, VSL, VCL, ALH, and BCF.
Newer versions of HT CASA software generate ASCII files that are organized differently and contain additional data. If you are using software versions 14.0 or earlier, save your ASCII files in the original .DBT format and download these for CASAnova. Later versions generate ASCII files in .txt (tab delimited) format. Save files in this .txt format for CASAnova. We modified CASAnova to ignore any "static" sperm included in the .txt files. Outputs from both .DBT and .txt files will be reported as percentages of the motile sperm population.
To generate motility profiles using CASAnova:
- Click "Start a Run".
- Click "Choose Files" and select the ASCII files to be analyzed. Open files to upload.
- Select whether you are analyzing mouse or human sperm.
- Click "Submit" (acknowledge that you agree to the license terms).
- Your analyzed data will appear on the page and can be downloaded in spreadsheet format by selecting the "Download summary spreadsheet" link.
Understanding data output:
Results are displayed as a table with the headings:
Filename, Tracks, Progressive, P%, Intermediate, I%, Hyperactivated, H%, Slow, S%, Weak, W%
Each row in the table corresponds to a different file uploaded to the program. Submitted file names are in the first column. The total number of motile tracks in each file is shown in the second column. The remaining column headings denote either the number of tracks (Progressive, Intermediate, etc.) or the percentage of tracks (P%, I%, etc.) in each file classified as the corresponding motility pattern. Typically, motility profiles are presented as the percentage of the total motile population undergoing each particular pattern of motility.
To see the classifications of individual tracks in a file, click on the filename and download the results file, which will be named "n_filename.cvs". The CASAnova classification for each track is shown in the first column.