ArticleNature communications2024
Automated neuronal reconstruction with super-multicolour Tetbow labelling and threshold-based clustering of colour hues.
Article in Nature communications, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- AAV-delivered TurboRFP enables streamlined tracing and analysis of corticospinal tract sprouting.Cell reports methods · 2026Article
- A CandiChrome toolkit for multicolor labeling ofbioRxiv : the preprint server for biology · 2026Article
- SegAnyNeuron: a neural image segmentation network with strong generalization performance by modeling image intensity variation.Brain informatics · 2026Article
- AAV-Based Bright and Sparse Labeling of Versatile Neurons Adaptable in Cre-Dependent Genetic Backgrounds.eNeuro · 2026Article
- Fluorescent Labeling Methods for Brain Structure Research.Molecules (Basel, Switzerland) · 2026Review
- Combinatorial protein barcodes enable self-correcting neuron tracing with nanoscale molecular context.bioRxiv : the preprint server for biology · 2025Article
- Historical trends in neuroanatomical tract-tracing techniques.Anatomical science international · 2025Review
- Live-cell omics with Raman spectroscopy.Microscopy (Oxford, England) · 2025Review
- Genetically Encoded Fluorescence Barcodes Allow for Single-Cell Analysis via Spectral Flow Cytometry.ACS synthetic biology · 2025Article
- Creating coveted bioluminescence colors for simultaneous multi-color bioimaging.Science advances · 2025Article
- A deep learning pipeline for morphological and viability assessment of 3D cancer cell spheroids.Biology methods & protocols · 2025Article
- Multi-color fluorescence live-cell imaging in Dictyostelium discoideum.Cell structure and function · 2024Article
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Authors and funding
8 authors.
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Abstract
Fluorescence imaging is widely used for the mesoscopic mapping of neuronal connectivity. However, neurite reconstruction is challenging, especially when neurons are densely labelled. Here, we report a strategy for the fully automated reconstruction of densely labelled neuronal circuits. Firstly, we establish stochastic super-multicolour labelling with up to seven different fluorescent proteins using the Tetbow method. With this method, each neuron is labelled with a unique combination of fluorescent proteins, which are then imaged and separated by linear unmixing. We also establish an automated neurite reconstruction pipeline based on the quantitative analysis of multiple dyes (QDyeFinder), which identifies neurite fragments with similar colour combinations. To classify colour combinations, we develop unsupervised clustering algorithm, dCrawler, in which data points in multi-dimensional space are clustered based on a given threshold distance. Our strategy allows the reconstruction of neurites for up to hundreds of neurons at the millimetre scale without using their physical continuity.
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