# Quickstart This quickstart provides a practical overview of the main workflows available in theatRICS. Depending on your experiment, you may use one or more of the following: - **RICS Export** - **RICS Fitting** - **SFCS** - **FCS Fitting** - **FRAP** - **Image Simulation** --- ## Start the program After installation, launch the GUI with: ```bash theatrics ``` --- ## Workflow 1: RICS analysis from image stacks Use this workflow when starting from microscopy image stacks such as TIFF or CZI data. ### Step 1 — Export RICS maps 1. Open the **RICS Export** tab. 2. Select an **input file** or **input folder**. 3. Choose: - channel - crop factor - window size - optional drift correction 4. Click **Export RICS**. This produces correlation maps and uncertainty maps. ### Step 2 — Fit the exported RICS map 1. Open the **RICS Fitting** tab. 2. Select the exported RICS map file. 3. Enter the microscope parameters: - pixel size - pixel dwell time - line time - PSF size - PSF aspect ratio 4. Choose a diffusion model. 5. Click **Run 2D/3D Fitting**. You can optionally enable: - **1D Fast Axis Fit** - **Diffusion map generation** ### Outputs Typical outputs include: - RICS correlation TIFF - uncertainty TIFF - fit summary CSV - NPZ fit arrays - SVG fit figures --- ## Workflow 2: SFCS analysis Use this workflow for perpendicular scanning FCS experiments. ### Steps 1. Open the **SFCS** tab. 2. Select an input file. 3. Choose the channel. 4. Optionally enable **Bleach Correction**. 5. Set the number of CPU cores. 6. Click **Correlate**. ### Display The GUI will show: - original data preview - aligned data preview - intensity trace - correlation curve ### Outputs Typical outputs include: - correlation curves - uncertainty estimates - exported SVG plots --- ## Workflow 3: FCS fitting from exported CSV curves Use this workflow when you already have correlation curves saved as CSV files. ### Single-file fitting 1. Open the **FCS Fitting** tab. 2. Select a **Single CSV** file. 3. Choose a fitting model. 4. Set: - tau range - PSF radius - PSF aspect ratio - experiment temperature 5. Adjust model-dependent initial parameters if needed. 6. Click **Run Fit**. ### Batch fitting 1. Select a **Batch folder**. 2. Set a **Batch file pattern**, for example: ```text *_xy_intensity_trace_correlation.csv ``` 3. Choose the model and fit parameters. 4. Click **Run Fit**. The software recursively searches matching files in subfolders. ### Calibration models For models whose names contain `Cal`, the GUI also shows: - **Given D** - **Given D temperature** These are hidden for non-calibration models. ### Outputs Per file: - SVG figure - fitted curve CSV - iMSD CSV where applicable For batch mode: - one summary CSV in the outer selected folder --- ## Workflow 4: FRAP analysis Use this workflow for fluorescence recovery after photobleaching data stored in CZI files with circular ROI annotations. ### Single-file analysis 1. Open the **FRAP** tab. 2. Select a **Single CZI** file. 3. Set: - imaging bleach correction - optional fallback pixel size - initial diffusion estimate - diffusion bounds 4. Click **Run FRAP**. ### Batch analysis 1. Select a **Batch folder**. 2. Set a pattern such as: ```text *FRAP*.czi ``` 3. Click **Run FRAP**. The software recursively processes matching FRAP files. ### Automatic FRAP processing The FRAP workflow automatically: - identifies the bleach frame - identifies the control ROI - normalizes traces with the control ROI - fits FRAP recovery curves ### Outputs Per file: - `*_FRAP_raw_data.xlsx` - `*_FRAP_summary.xlsx` - `*_FRAP_overview.svg` --- ## Workflow 5: Simulated data generation Use this workflow to generate synthetic image stacks for testing and validation. ### Steps 1. Open the **Image Simulation** tab. 2. Set: - image size - number of frames - number of particles - diffusion coefficients - simulation type - output file path 3. Click **Run Simulation**. ### Outputs The GUI shows: - first and last frame - average projection - intensity trace The simulation is also saved as a TIFF stack. --- ## Workflow 6: Diffusion map generation Diffusion maps are generated from the **RICS Fitting** tab. ### Steps 1. Open **RICS Fitting**. 2. In the **Diffusion Map Fitting Parameters** section: - select an input file - choose channel - set window size - set offset 3. Click **Generate Diffusion Map**. ### Outputs Typical outputs include: - diffusion map - brightness map - number map - auxiliary result arrays --- ## Monitoring progress During long-running jobs, the GUI provides: - a **status bar** - a **progress bar** - a **Cancel Running Task** button - textual logs in the **Results & Logs** tab For large batch jobs, the display may update only after the current file or at the end of the batch to keep the interface responsive. --- ## Saving and exporting The **Results & Logs** tab allows you to: - save log output - export plots - save a GUI session - reload a previous session Plots are usually exported in high-quality formats such as SVG, while numerical outputs are saved as CSV, NPZ, TIFF, or Excel files depending on the workflow. --- ## Suggested starting points ### If you work with image stacks Start with: - **RICS Export** - then **RICS Fitting** ### If you work with scan correlation curves Start with: - **SFCS** - then optionally **FCS Fitting** ### If you work with photobleaching recovery experiments Start with: - **FRAP** ### If you want to test the pipeline Start with: - **Image Simulation**