Welcome to the Syn2Ves Program Documentation! 🎉

Syn2Ves is a program designed to pair synapses with vesicles based on position, and quantify the alignment of these pairings.

The program takes the following inputs:

While the software can be used on it's own, Syn2Ves was intended to be used as a part of the ORS dragonfly software. This section will give a brief overview of how to install all of the neccessary dragonfly files in order to have it run correctly.

All Dragonfly Files to download are in the “Dragonfly” folder.

1. Download all of the files in the Menu Items folder
2. Click Windows Key + R to open the run command
3. Type %AppData% and click OK
4. Go to Local\ORS\Dragonfly(newest version)\pythonUserExtensions\GenericMenuItems
5. In this folder, drag in all of the downloaded files (Put each of the files into here individually, not the entire folder)
6. Open the config.ini file, and set the paths to the appropriate locations (make sure all slashes are back slashes)

1. Download all of the files in the Macro folder
2. Click Windows Key + R to open the run command
3. Type %AppData% and click OK
4. Go to Local\ORS\Dragonfly(newest version)\pythonUserExtensions\Macros
5. In this folder, drag in the .py file

1. Select 1 or more MultiROIs that you'd like to fully export in the Properties tab on Dragonfly
2. Run the Menu Item
3. Select where you'd like your meshes exported (Each MultiROI will get its own folder within the directory you choose)

1. Run the Menu Item to launch the program
2. Firstly, select the two mesh folders. These are the exports of the "Export MultiROI Labels as Meshes" menu we just ran
3. Select your Synapse and Vesicle CSVs, which are also exports of the previous menu
4. Run Pairing
5. Select the "paired" CSV exported from the Pairing workflow as your Pairing CSV for Alignment Analysis
6. Run Alignment Analysis

A folder containing STL files of synapses, named numerically (example: 1.stl, 2.stl, etc). The program will only consider files with the ".stl" extension.

A folder containing STL files of vesicles, named numerically (example: 1.stl, 2.stl, etc). The program will only consider files with the ".stl" extension.

A CSV file containing the pairing information of the synapses and vesicles. The file must be formatted with the following column names: synLabel and vesLabel. These columns should contain numerical labels for each synapse and vesicle, respectively.

A CSV file containing center of mass data for the synapses. The file must be formatted with the following column names: labels, comX, comY, comZ, vol, SA, halfSA, sphericity, maxFeretLength, minFeretLength, aspectRatio. These columns should contain numerical labels for each synapse, followed by its center of mass coordinates (x, y, z), volume, surface area, half surface area, sphericity, maximum Feret length, minimum Feret length, and aspect ratio, respectively.

Same as Synapse CSV but for Vesicles.

The search radius from a synapses' center of mass. The program will only pair a vesicle cloud with a synapse if its center of mass falls within the cubic volume with this radius.

To run the program, clone this repository and execute the following commands:

cd Syn2Ves-User

pip install -r requirements.txt

python -u main.py

We are using pyinstaller to compile Syn2Ves into a finished application. The steps differ based on your platform, but the following instructions are for windows (10-11).

First, ensure you have a main.spec and resources.qrc file. If you don't, run the following commands:

pyinstaller main.spec

pyrcc5 resources.qrc -o resources.py

Then to compile to exe enter the directory with main.py and run the following command:

WINDOWS:

pyinstaller.exe --onefile --windowed --icon=logo.ico main.py

MACOS:

pyinstaller --onefile --windowed --icon=logo.ico --codesign-identity=YOUR_APPLE_DEVELOPER_CERT_CODESIGN_IDENTITY --target-arch [x86_64|universal2] main.py

This will spew output in the console and may take a while. The final exe file can be found in the /dist directory.

I recommend reading through this handy article for more details regarding the specifics of compilation, particularly if you're having trouble getting icons to load: Packaging PyQt5 applications for Windows, with PyInstaller

Pyinstaller should be capable of building for MacOS, but I have yet to test it. If you're trying to compile for Linux, you're capable of figuring out how (or you can just use wine).

Outputs a CSV file named syn2ves_output.csv containing the following columns:

• synLabel: numerical label of the synapse
• vesLabel: numerical label of the vesicle
• Og_Ves_X: x value of the original vesicle position in nm
• Og_Ves_Y: y value of the original vesicle position in nm
• Og_Ves_Z: z value of the original vesicle position in nm
• Ves_X: x value of the rotated vesicle position in nm
• Ves_Y: y value of the rotated vesicle position in nm
• Ves_Z: z value of the rotated vesicle position in nm
• Syn_X: x value of the synapse position in nm
• Syn_Y: y value of the synapse position in nm
• Syn_Z: z value of the synapse position in nm
• VectorAngle: the vector angle, in degrees, between the center of mass of the original vesicle (Og_Ves) and the center of mass of the maximum surface area adjusted vesicle (Ves), with the synapse center of mass (Syn) at the center
• Intersect: a value representing the amount of intersecting pixels when projections of the synapse and vesicle are aligned
• IOU: intersection over union of the synapse and vesicle projections
• IOS: intersection over synapse area of the synapse and vesicle projections