dropdrop 2.0.0

Automated pipeline for detecting droplets and inclusions in microscopy images, powered by Cellpose

DropDrop

Automated pipeline for detecting droplets and inclusions (beads) in microscopy z-stacks using Cellpose segmentation and morphological analysis.

Tailored for the EVOS M5000 Imaging System.

Installation

# Using uv (recommended)
uv pip install dropdrop

# Using pip
pip install dropdrop

From source

git clone https://github.com/yourusername/dropdrop.git
cd dropdrop
uv pip install -e .

Quick Start

# Single directory — interactive prompts for settings
dropdrop ./images

# Process only first 5 frames (for testing)
dropdrop ./images -n 5

# With interactive editor
dropdrop ./images -e

# Multiplex mode — batch process subdirectories
dropdrop -m ./samples

Usage

Single Mode

# Basic run (prompts for settings interactively)
dropdrop ./images

# Custom output directory
dropdrop ./images ./results/my_project

# With editor and archive
dropdrop ./images -e -z

Multiplex Mode

Process multiple sample directories at once. Each subdirectory is labeled interactively and processed as a separate sample, then combined into a multiplexed report.

dropdrop -m ./samples_parent_dir
dropdrop -m ./samples_parent_dir -z    # Archive result

Resume (Resurrect)

If a multiplex run is interrupted, resume from where it left off:

dropdrop -r

Cache Control

dropdrop ./images --no-cache       # Disable caching
dropdrop ./images --clear-cache    # Clear cache before run

Interactive Editor

The editor (-e) allows manual correction of detected inclusions:

Key	Action
Left-click	Add inclusion
Right-click (hold)	Remove inclusions
s	Toggle droplet selection (hover over droplet)
u	Undo last action
c	Clear all inclusions in frame
d	Toggle droplet visibility
Arrow keys / Space	Navigate frames
q / Esc	Exit

Disabled droplets (gray with X) are excluded from the final results.

Output Structure

Single mode

results/<YYYYMMDD>_<label>/
  data.csv                  # Raw detection data
  summary.txt               # Settings and statistics
  report.png                # Combined report with sample frames
  size_distribution.png     # Droplet diameter histogram
  poisson_comparison.png    # Bead distribution vs theoretical

Multiplex mode

results/<YYYYMMDD>_multiplex/
  data.csv                  # Merged data with sample column
  summary.txt               # Per-sample statistics
  summary_report.png        # Comparison table, overlaid plots, sample collage
  size_distribution.png     # Overlaid diameter histograms
  poisson_comparison.png    # Overlaid inclusion distributions

data.csv columns

Column	Description
sample	Sample label (multiplex only)
frame	Frame index
droplet_id	Droplet ID within frame
center_x, center_y	Droplet center coordinates (px)
diameter_px, diameter_um	Droplet diameter
area_px, area_um2	Droplet area
inclusions	Number of inclusions detected

Architecture

CLI
  -> Detection (per sample) -> .tmp_<label>/data.csv + sample_*.png
  -> Analysis.run(output_dir)  -- auto-discovers .tmp_* dirs
       1 sample  -> single report
       2+ samples -> multiplex report
  -> Cleanup .tmp_* dirs
  -> Archive (optional)

Configuration

Create config.json in your working directory to customize detection parameters:

{
  "cellpose_flow_threshold": 0.4,
  "cellpose_cellprob_threshold": 0.0,
  "erosion_pixels": 5,
  "kernel_size": 7,
  "tophat_threshold": 30,
  "min_inclusion_area": 7,
  "max_inclusion_area": 50,
  "edge_buffer": 5,
  "min_droplet_diameter": 80,
  "max_droplet_diameter": 200,
  "px_to_um": 1.14,
  "cache": {
    "enabled": true,
    "max_frames": 100
  }
}

Parameters

Parameter	Description
cellpose_flow_threshold	Cellpose flow threshold for segmentation
cellpose_cellprob_threshold	Cellpose cell probability threshold
erosion_pixels	Pixels to erode droplet mask before inclusion detection
kernel_size	Morphological kernel size for black-hat transform
tophat_threshold	Threshold for inclusion detection
min/max_inclusion_area	Inclusion size constraints (px)
edge_buffer	Buffer from image edge to ignore inclusions
min/max_droplet_diameter	Droplet size constraints (px)
px_to_um	Pixel to micrometer conversion factor

Requirements

• Python 3.12+
• CUDA-capable GPU (recommended for Cellpose)

License

MIT