resolutiontree 0.2

Systematic exploration of clustering resolutions in single-cell analysis

ResolutionTree

Systematic exploration of clustering resolutions in single-cell analysis

Overview

ResolutionTree is a Python package that automates the exploration of different clustering resolutions in single-cell RNA sequencing data. Instead of manually testing different resolution parameters, ResolutionTree systematically evaluates how subclusters emerge from parent clusters as resolution increases, providing a hierarchical view of cluster relationships along with key differentially expressed genes (DEGs) that distinguish them.

The Problem

One of the most common questions in single-cell analysis is: "How do you decide on the clustering resolution?" The reality is:

• There's no universal "golden standard" for selecting the best resolution
• The choice depends on your specific biological question
• Traditional approaches require manual iteration through multiple resolutions
• The process is time-consuming and subjective

The Solution

ResolutionTree provides:

• Automated resolution exploration across user-defined ranges
• Hierarchical cluster visualization showing parent-child relationships
• Differential gene expression analysis highlighting key distinguishing features
• Customizable visualization options for publication-ready figures
• Integration with Scanpy workflow

Installation

From GitHub Release (Recommended)

pip install resolutiontree==0.2

From Source

pip install git+https://github.com/joe-jhou2/resolutiontree.git

Dependencies

pip install scanpy pandas numpy scipy matplotlib seaborn networkx igraph-python

Quick Start

import scanpy as sc
import resolutiontree as rt

# Load your data
adata = sc.datasets.pbmc3k()

# Standard preprocessing
sc.pp.normalize_total(adata, inplace=True)
sc.pp.log1p(adata)
sc.pp.pca(adata)
sc.pp.neighbors(adata)
sc.tl.umap(adata)

# Define resolutions to explore
resolutions = [0.0, 0.2, 0.5, 1.0, 1.5, 2.0]

# If you don't want to modify the original AnnData object, make a copy first
adata_new = adata.copy()

# Step 1: Find optimal resolution with DEG analysis
rt.cluster_resolution_finder(adata_new,
                             resolutions=resolutions,
                             n_top_genes=3,
                             min_cells=2,
                             deg_mode="within_parent"
                             )

# Step 2: Visualize the hierarchical clustering tree
rt.cluster_decision_tree(adata_new, resolutions=resolutions, 
                        output_settings = {
                            "output_path": "result.png",
                            "draw": False,
                            "figsize": (12, 6),
                            "dpi": 300
                            },
                        node_style = {
                            "node_size": 500,
                            "node_colormap": None,
                            "node_label_fontsize": 12
                            },
                        edge_style = {
                            "edge_color": "parent",
                            "edge_curvature": 0.01,
                            "edge_threshold": 0.01,
                            "show_weight": True,
                            "edge_label_threshold": 0.05,
                            "edge_label_position": 0.8,
                            "edge_label_fontsize": 8
                            },
                        gene_label_settings = {
                            "show_gene_labels": True,
                            "n_top_genes": 2,
                            "gene_label_threshold": 0.001,
                            "gene_label_style": {"offset":0.5, "fontsize":8},
                            },
                        level_label_style = {
                            "level_label_offset": 15,
                            "level_label_fontsize": 12
                            },
                        title_style = {
                            "title": "Hierarchical Leiden Clustering",
                            "title_fontsize": 20
                            },
                        layout_settings = {
                            "node_spacing": 5.0,
                            "level_spacing": 1.5
                            },
                        clustering_settings = {
                            "prefix": "leiden_res_",
                            "edge_threshold": 0.05
                            }
                    )

Key Features

1. Resolution Finding

The cluster_resolution_finder() function systematically tests multiple clustering resolutions and identifies key differentially expressed genes:

DEG Comparison Modes:

• "within_parent": Compare sibling clusters from the same parent (recommended)
• "per_resolution": Compare all clusters at each resolution

2. Hierarchical Visualization

The cluster_decision_tree() function creates publication-ready hierarchical cluster trees:

3. Customization Options

The detailed customization options allow you to tailor the visualization to your needs: see my blog: Optimizing Clustering Resolution in Single-Cell Analysis

Example Output

The package generates hierarchical trees showing:

1. Cluster evolution across resolutions
2. Parent-child relationships between clusters
3. Key differentiating genes for each split
4. Cluster sizes and transition weights

Use Cases

Fine-grained Cell Type Discovery

• T cell subsets: Higher resolutions reveal distinct T helper, regulatory, and cytotoxic populations
• Immune cell states: Identify activation states and differentiation trajectories
• Rare cell populations: Detect small but biologically important cell clusters

Quality Control

• Over-clustering detection: Identify when resolution becomes too high
• Biological validation: Use DEGs to confirm cluster biological relevance
• Resolution optimization: Find the sweet spot between under- and over-clustering

Integration with Scanpy

ResolutionTree seamlessly integrates with standard Scanpy workflows:

# Standard Scanpy preprocessing
sc.pp.filter_cells(adata, min_genes=200)
sc.pp.filter_genes(adata, min_cells=3)
sc.pp.normalize_total(adata, target_sum=1e4)
sc.pp.log1p(adata)
sc.pp.highly_variable_genes(adata)
sc.pp.pca(adata)
sc.pp.neighbors(adata)

# ResolutionTree analysis
rt.cluster_resolution_finder(adata, resolutions=[0.2, 0.5, 1.0, 1.5])
rt.cluster_decision_tree(adata, resolutions=[0.2, 0.5, 1.0, 1.5])

# Continue with Scanpy
sc.tl.umap(adata)
sc.pl.umap(adata, color='leiden_res_1.0')

Citation

If you use ResolutionTree in your research, please cite:

@software{resolutiontree2025,
  author = {Joe Hou},
  title = {ResolutionTree: Systematic exploration of clustering resolutions in single-cell analysis},
  url = {https://github.com/joe-jhou2/resolutiontree},
  year = {2025}
}

Blog Post

For a detailed explanation of the methodology and examples, see the blog post: Optimizing Clustering Resolution in Single-Cell Analysis

Contributing

Contributions are welcome! Please feel free to submit a Pull Request. For major changes, please open an issue first to discuss what you would like to change.

License

This project is licensed under the MIT License - see the LICENSE file for details.

Changelog

v0.1.0 (2025-06-03)

• Initial release
• Core resolution finding functionality
• Hierarchical cluster tree visualization
• Scanpy integration
• Comprehensive documentation and examples

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Keywords: single-cell RNA-seq, clustering, resolution selection, differential gene expression, hierarchical clustering, scanpy, bioinformatics