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🌌 Spatial Transcriptomics Tools

Other resourceawesome_spatial_omics

Table of Contents

General Tools

  • Best practices Bioconductor - [R] - Principles for statistical analysis of spatial transcriptomics data
  • squidpy - [Python] - Spatial single cell analysis toolkit from scverse
  • Giotto - [R/Python] - Comprehensive spatial data analysis suite
  • Vitessce - [JavaScript] - Visual integration tool for exploration of spatial single cell experiments
  • Voyager - [R] - Spatial transcriptomics visualization from Pachter lab
  • BASS - [R] - Multiple sample analysis
  • SpaVAE - [Python] - All-purpose tool for dimension reduction, visualization, clustering, batch integration, denoising, differential expression, spatial interpolation, and resolution enhancement
  • sopa - [Python] - Spatial omics processing and analysis
  • SpatialAgent - [Python] - An autonomous AI agent for spatial biology
  • ChatSpatial - [Python] - MCP server enabling spatial transcriptomics analysis via natural language, integrating 60+ methods including SpaGCN, Cell2location, LIANA+, CellRank for Visium, Xenium, MERFISH | Docs | PyPI
  • LazySlide - [Python] - Framework for whole slide image (WSI) analysis
  • pasta - [R] - Point pattern and lattice data analysis from Robinson lab
  • rakaia - [JavaScript] - Scalable interactive visualization and analysis of spatial omics including spatial transcriptomics, in the browser (Website)
  • semla - [R] - Useful tools for Spatially Resolved Transcriptomics data analysis and visualization
  • sosta - [Python] - Spatial Omic Structure Analysis
  • SPATA2 - [R] - Spatial transcriptomics analysis toolkit
  • Thor - [Python] - Comprehensive platform for cell-level analysis with anti-shrinking Markov diffusion and 10 modular tools paired with Mjolnir web interface
  • VR-Omics - [GUI] - Free platform-agnostic software with end-to-end automated processing of multi-slice spatial transcriptomics data through biologist-friendly GUI | Windows | MacOS | GitHub
  • CosMx-Analysis-Scratch-Space - [R/Python] - Analysis resources and tools for CosMx SMI spatial transcriptomics | GitHub
  • SpaceSequest - [R] - Unified pipeline for analysis, visualization, and publication of spatial transcriptomics data from Visium, Visium HD, Xenium, GeoMx, and CosMx | Tutorial

Nextflow / Pipelines

Analysis Pipeline Steps

ROI Selection

  • S2Omics - [Python] - Designing smart spatial omics experiments with S2Omics

QC

  • SpaceTrooper - [R] - Quality control for spatial transcriptomics
  • GrandQC - [Python] - Comprehensive solution for quality control in digital pathology
  • SpotSweeper - [R] - Spatially aware quality control for spatial transcriptomics
  • MerQuaCo - [Python] - A computational tool for quality control in image-based spatial transcriptomics

Normalization

  • Cell volume normalization - [R] - Recommended for imaging-based techniques, especially with small probe lists
  • SpaNorm - [R] - First spatially-aware normalization method that concurrently models library size effects and underlying biology | Bioconductor

Gene Imputation & Denoising

  • Note: Gene imputation is not recommended for deconvolution tasks
  • SpaGE - [Python] - Spatial gene expression prediction with best overall performance
  • SpaGCN - [Python] - Spatial graph convolutional network for gene correlation analysis
  • Tangram - [Python] - Transcript distribution prediction and spatial mapping
  • SpaOTsc - [Python] - Spatial imputation via optimal transport
  • Seurat integration workflow - [R] - Transfer gene expression from scRNA-seq reference
  • Sprod - [Python] - Spatial denoising method
  • TISSUE - [Python] - Transcript imputation with spatial single-cell uncertainty estimation
  • SpaIM - [Python] - Single-cell Spatial Transcriptomics Imputation via Style Transfer

Bias Correction

  • ResolVI - [Python] - Bias correction method
  • Statial - [R] - Correction of spill-over effects
  • ovrl.py - [Python] - A python tool to investigate vertical signal properties of imaging-based spatial transcriptomics data
  • SPLIT - [R] - SPLIT effectively resolves mixed signals and enhances cell-type purity
  • cellAdmix - [R] - From Kharchenko lab - Evaluating and correcting cell admixtures in imaging-based spatial transcriptomics data.
  • DenoIST - [R] - Denoising Image-based Spatial Transcriptomics data
  • MisTIC - [Python] - A probabilistic model for correcting mis-assigned transcripts due to cell segmentation errors

Cell Segmentation

Imaging-based Segmentation

  • Baysor - [Julia] - Bayesian segmentation of spatial transcriptomics data
  • Cellpose - [Python] - Generalist algorithm for cellular segmentation
    • Cellpose 3 - With supersampling/restoration capabilities
    • Cellpose-SAM - Cell and nucleus segmentation with superhuman generalization, works in 3D with various image conditions
  • DeepCell - [Python] - Deep learning library for single cell analysis
  • Bo Wang's method - [Python] - Better than SOTA segmentation (Nature Methods 2024)
  • Proseg - [Rust] - Probabilistic segmentation method
  • ComSeg - [Python] - Transcript-based point cloud segmentation
  • FICTURE - [Python] - Feature-based image segmentation
  • Xenium cell boundary - [Web] - Alternative when interior staining fails
  • Bioimage.io - [Web] - Repository of AI models for segmentation
  • ST-cellseg - [Python] - Segmentation for spatial transcriptomics
  • CelloType - [Python] - Cell type detection and segmentation
  • SAINSC - [Python] - Segmentation for sequencing-based spatial data
  • BIDCell - [Python] - Biologically-informed deep learning for subcellular spatial transcriptomics segmentation
  • FastReseg - [R] - Using transcript locations to refine image-based cell segmentation results
  • Segger - [Python] - Fast and accurate cell segmentation of imaging-based spatial transcriptomics data
  • Bering - [Python] - Graph deep learning for joint noise-aware cell segmentation and molecular annotation in 2D and 3D spatial transcriptomics
  • STP - [Python] - Single-cell Partition for subcellular spatially-resolved transcriptomics integrating data with nuclei-stained images
  • Deep learning-based segmentation - [Python] - Extensively trained nuclear and membrane segmentation models for precise transcript assignment in CosMx SMI data
  • CellSAM - [Python] - Foundation model for cell segmentation achieving state-of-the-art performance across cellular targets (bacteria, tissue, yeast, cell culture) and imaging modalities (brightfield, fluorescence, phase, multiplexed) | Paper | Web App

Segmentation-free methods:

  • SSAM - [Python] - Subcellular segmentation-free analysis by multidimensional mRNA density
  • Points2Regions - [Python] - Transcript-based region identification without segmentation

VisiumHD Segmentation

  • Bin2Cell - [Python] - Segmentation for VisiumHD data
  • ENACT - [Python] - Enhanced accuracy for VisiumHD segmentation
  • STHD - [Python] - Cell annotation for VisiumHD

Cell Annotation

  • STEM - [Python] - Cell type annotation method
  • TACIT - [Python] - Automated cell type identification
  • moscot - [Python] - Optimal transport-based cell mapping
  • CELLama - [Python] - Cell annotation model
  • TACCO - [Python] - Transfer of annotations between single-cell datasets
  • TANGRAM - [Python] - Mapping single-cell to spatial data
  • MMoCHi - [Python] - Cell annotation method
  • CytoSPACE - [Python] - High-resolution alignment of single-cell and spatial transcriptomes
  • ABCT - [R] - Anchor-based Cell Typer
  • STHD - [Python] - Cell annotation for VisiumHD
  • STELLAR - [Python] - Annotation of spatially resolved single-cell data with STELLAR
  • Vesalius - [R] - Multi-scale and multi-context interpretable mapping of cell states across heterogeneous spatial samples
  • STALocator - [Python] - ST-Aided Locator using deep learning to localize cells from single-cell RNA-seq data onto tissue slices
  • CMAP - [Python] - Cellular Mapping of Attributes with Position, maps large-scale individual cells to precise spatial locations using divide-and-conquer strategy
  • TransST - [Python] - Transfer learning framework leveraging cell-labeled information from external sources for cell-level heterogeneity inference | GitHub
  • InSituType - [R] - Cell typing for CosMx SMI spatial transcriptomics
  • HieraType - [R] - Hierarchical cell typing using RNA + protein for CosMx SMI
  • CosMx-Cell-Profiles - [R] - Collection of reference datasets for CosMx SMI
  • GARDEN - [Python] - Graph-based dynamic attention framework for identifying rare pathogenic cell populations (disease-driving cells often missed by standard methods), enables 3D tissue reconstruction

Cell Deconvolution

  • RCTD - [R] - Robust cell type decomposition
  • Cell2location - [Python] - Mapping scRNA-seq to spatial data
  • SPOTlight - [R] - Seeded NMF regression to deconvolute spatial spots
  • CARD - [R] - Spatially informed cell-type deconvolution
  • FlashDeconv - [R] - High-performance deconvolution using randomized sketching, achieves linear O(N) scaling for Visium HD and other high-resolution platforms

Differential Expression

  • C-SIDE - [R] - Cell type-Specific Inference of Differential Expression in spatial transcriptomics
  • Niche-DE - [R] - Niche-differential gene expression analysis identifying context-dependent cell-cell interactions
  • smiDE - [R] - Spatial differential expression method | GitHub
  • spatialGE - [R] - Spatial gene expression analysis
  • Vespucci - [R] - Prioritize spatial regions involved in the response to an experimental perturbation in spatial transcriptomics
  • CSDE - [Python] - Corrected Spatial Differential Expression using Prediction-Powered Inference to account for preprocessing uncertainties (segmentation, quantification, cell typing)

Spatially Variable Genes

  • PROST - [Python] - Detection of spatially variable genes
  • SpatialDE - [Python] - Spatial differential expression analysis
  • SPARK-X - [R] - Detection of spatially variable genes, best performing
  • Hotspot - [Python] - Identify informative gene modules with lowest false positive rate
  • SOMDE - [Python] - Self-organizing map for spatially variable gene detection with optimization
  • trendsceek - [R] - Identification of spatial expression trends
  • nnSVG - [R] - Scalable identification of spatially variable genes using nearest-neighbor Gaussian processes
  • SLOPER - [Python] - Score-based learning of Poisson-modeled expression rates for spatial gene modules and tissue organization patterns

Integration

  • PRECAST - [R] - Probabilistic embedding, clustering, and alignment for integrating spatial transcriptomics data
  • MISO - [Python] - MultI-modal Spatial Omics for versatile feature extraction and clustering integrating multiple modalities including gene expression, protein, epigenetics, metabolomics, and histology
  • SIMO - [Python] - Spatial Integration of Multi-Omics through probabilistic alignment integrating spatial transcriptomics with multiple single-cell modalities
  • GSI - [Python] - Gene Spatial Integration using deep learning with representation learning to extract spatial distribution of genes | GitHub | Zenodo
  • SPACE-seq - [Paper] - Unified molecular approach for spatial multiomics enabling simultaneous analysis of chromatin accessibility, mitochondrial DNA mutations, and gene expression on standard 10× Genomics Visium CytAssist platform
  • LLOKI - [Python] - Cross-platform spatial transcriptomics integration using optimal transport and scGPT foundation models for unified features across different gene panels (RECOMB 2025)

Cell Niches & Tissue Domains

(Smaller) Cell types → Cell modules/neighborhoods → Niches/tissue domains (Larger)

  • BANKSY - [R/Python] - Unified cell typing and tissue domain segmentation
  • CellCharter - [Python] - Hierarchical niche detection
  • SpatialGLUE - [Python] - Multi-omics cell niche identification
  • smoothclust - [R] - Spatial clustering
  • SpaTopic - [R] - Spatial topic modeling
  • hdWGCNA - [R] - Weighted gene correlation network analysis
  • GASTON - [Python] - Graph-based spatial domain detection
  • SpatialMNN - [R] - Identification of shared niches between slides
  • NicheCompass - [Python] - End-to-end analysis of spatial multi-omics data
  • Proust - [Python] - Spatial domain detection using contrastive self-supervised learning for spatial multi-omics technologies (multi-modal domains)
  • STAMP - [Python] - Spatial Transcriptomics Analysis with topic Modeling, provides interpretable dimension reduction through deep generative modeling discovering tissue domains and cellular communication patterns
  • DeepGFT - [Python] - Combines deep learning with graph Fourier transform for spatial domain identification | GitHub | Zenodo
  • novae - [Python] - Deep learning framework for spatial domain detection and tissue organization analysis

Cell Distances & Neighborhood

  • CRAWDAD - [R] - Cell relationship analysis with directional adjacency distributions
  • HoodscanR - [R] - Neighborhood analysis
  • SpicyR - [R] - Spatial analysis in R
  • MISTy - [R] - Explainable multiview framework for dissecting spatial relationships from highly multiplexed data
  • SpatialCorr - [Python] - Identifying gene sets with spatially varying correlation structure
  • CatsCradle - [R] - Spatial analysis framework for tissue neighbourhoods

Spatial Trajectories

  • spaTrack - [Python] - Spatial trajectory analysis
  • scSpace - [Python] - Reconstruction of cell pseudo-space from single-cell RNA sequencing data
  • SOCS - [Python] - Accurate trajectory inference in time-series spatial transcriptomics with structurally-constrained optimal transport
  • STORIES - [Python] - Spatiotemporal Reconstruction Using Optimal Transport for cell trajectory inference from spatial transcriptomics profiled at multiple time points

Cell-Cell Communication

  • Spatia - [Python] - Spatial cell-cell interaction analysis
  • CellAgentChat - [Python] - Agent-based cell communication modeling
  • SpaTalk - [R] - Knowledge-graph-based cell-cell communication inference
  • SpaOTsc - [Python] - Inferring spatial and signaling relationships between cells
  • MISTy - [R] - Explainable multi-view framework for dissecting intercellular signaling
  • DeepLinc - [Python] - De novo reconstruction of cell interaction landscapes
  • CellChat - [R] - Inferrence of cell-cell communication from multiple spatially resolved transcriptomics datasets
  • COMMOT - [Python] - Screening cell-cell communication in spatial transcriptomics via collective optimal transport
  • NicheNet - [R] - Linking ligands to downstream target gene regulation
  • DeepTalk - [Python] - Single-cell resolution cell-cell communication using deep learning
  • CellNEST - [Python] - Cell–cell relay networks using attention mechanisms on spatial transcriptomics
  • FlowSig - [Python] - Inferring pattern-driving intercellular flows from single-cell and spatial transcriptomics

Metacells & Scalability

  • SuperSpot - [R] - Metacell analysis for spatial data
  • SEraster - [R] - Rasterization method for spatial data processing

Subcellular Analysis

  • Sprawl - [Python] - Subcellular transcript localization
  • Bento - [Python] - Python toolkit for subcellular analysis of spatial transcriptomics data
  • FISHfactor - [Python] - Analysis of subcellular transcript patterns
  • InSTAnT - [Python] - Intracellular spatial transcript analysis
  • troutpy - [Python] - Analysis of transcripts outside segmented cells in spatial transcriptomics data

Copy Number Variations

  • CalicoST - [Python] - CNV detection in spatial data
  • inSituCNV - [Python] - Inference of Copy Number Variations in Image-Based Spatial Transcriptomics

Isoform Analysis

  • SPLISOSM - [Python] - Spatial isoform statistical modeling for detecting isoform-resolution patterns (alternative splicing, polyadenylation) from spatial transcriptomics data | Paper | Docs

Transcription Factors & Gene Regulatory Networks

  • STAN - [R] - Spatial transcription factor analysis

Technical Enhancements

Slide Alignment

  • PASTE/PASTE2 - [Python] - Probabilistic alignment of spatial transcriptomics experiments
  • SPIRAL - [R] - Integrating and aligning spatially resolved transcriptomics data across different experiments, conditions, and technologies
  • TOAST - [Python] - Topography Aware Optimal Transport for Alignment of Spatial Omics Data
  • STalign - [Python] - Alignment of spatial transcriptomics data using diffeomorphic metric mapping
  • SANTO - [Python] - A coarse-to-fine alignment and stitching method for spatial omics

Super Resolution

  • TESLA - [Python] - Super resolution for 10X Visium
  • istar - [Python] - Super resolution for Visium
  • BayesSPACE - [R] - Subspot resolution
  • Spotiphy - [Python] - Super resolution tool for spatial data

Transcripts + Histology

  • ST-Net - [Python] - Integrating spatial gene expression and tumor morphology via deep learning
  • SpaceDIVA - [Python] - Integration of transcript data with histological images
  • HEST - [Python] - Dataset for Spatial Transcriptomics and Histology Image Analysis
  • CellLENS - [Python] - Cell Local Environment Neighborhood Scan
  • DeepSpot - [Python] - Leveraging Spatial Context for Enhanced Spatial Transcriptomics Prediction from H&E Images
  • SpotWhisperer - [Python] - Molecularly informed analysis of histopathology images using natural language
  • STPath - [Python] - A Generative Foundation Model for Integrating Spatial Transcriptomics and Whole Slide Images
  • AESTETIK - [Python] - AutoEncoder for Spatial Transcriptomics Expression with Topology and Image Knowledge

Benchmarks

Datasets & Foundation Models

Datasets

  • HISSTA - [Python/R] - Histopathology spatial transcriptomics dataset
  • STOmicsDB - [Web] - Spatial transcriptomics database
  • HistAI Pathology Datahub - [Python] - Skills repo / HistAI Whole Slide Image Data Hub

Foundation Models

  • KRONOS - [Python] - Foundation Model for Multiplex Spatial Proteomic Images
  • scGPT-spatial - [Python] - Language model for spatial transcriptomics
  • Phikon-v2 - [Python] - Spatial biology foundation model
  • Bioptimus H-optimus-0 - [Python] - Biology-focused foundation model
  • Bioptimus H-optimus-1 - [Python] - Latest biology-focused foundation model from Bioptimus
  • DeepCell dataset - [Web] - CNN + human features embeddings
  • TITAN - [Python] - A multimodal whole-slide foundation model for pathology
  • Virchow - [Python] - Foundation model for computational pathology
  • UNI and UNI2 - [Python] - Universal pathology foundation models
  • CONCH - [Python] - Contrastive learning for histopathology
  • GIGApath - [Python] - Large-scale pathology foundation model
  • OmiCLIP - [Python] - A visual–omics foundation model to bridge histopathology with spatial transcriptomics
  • Nicheformer - [Python] - Transformer-based foundation model pretrained on SpatialCorpus-110M containing over 110 million cells for spatial composition and label prediction | GitHub
  • FOCUS - [Python] - Foundational generative model for cross-platform unified ST enhancement conditioned on H&E images, scRNA-seq references, and spatial co-expression priors (trained on 1.7M H&E-ST pairs, 10 platforms)
  • Atlas 2 - [Python] - Foundation models for clinical deployment

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