Skeleton node reference¶

Start with the skeleton workflow tutorial to choose 2D or 3D processing and inspect your segmentation. This page lists the outputs, units, and assumptions needed to interpret the resulting tables.

Skeletonize¶

Input: binary mask.
Output: binary skeleton mask.
Purpose: Converts foreground objects into one-pixel/voxel-wide centerlines.
Spatial processing: 3D ZYX thins each complete ZYX volume once. Leading non-spatial axes are independent blocks, so TZYX runs one ZYX volume per timepoint. 2D YX thins each YX plane independently.
Auto from axes: explicit ZYX selects volumetric 3D processing and explicit YX selects 2D processing. A generic QYX stack is ambiguous and is rejected until Image Source records a reviewed declaration such as QYX -> ZYX, or the user deliberately chooses 2D YX.
Methods: Auto resolves deterministically to Lee for 3D and Zhang for 2D. Lee is valid for 2D and 3D. Zhang 2D is never valid for a 3D block, including an empty volume.
Algorithm contract: the 3D path uses scikit-image's implementation of the Lee, Kashyap, and Chu volumetric thinning algorithm. Lee examines a 3x3x3 neighbourhood and rechecks deletion candidates to preserve connectivity. Each processed block is surrounded by background at its boundary. The output preserves input shape, axis records, calibration, and leading-axis identity.
Provenance: the output metadata history records the actual Lee or Zhang method, resolved 2D/3D block scope, neighbourhood, and boundary assumption.
Use before: Analyze Skeleton, Measure Skeleton Branches, Skeleton Keypoints, Skeleton Graph Overlay, Label Skeleton Components, Label Skeleton Branches, and Prune Skeleton Branches.

The Lee implementation follows T.-C. Lee, R. L. Kashyap, and C.-N. Chu, "Building skeleton models via 3-D medial surface/axis thinning algorithms", Computer Vision, Graphics, and Image Processing 56(6), 1994. VIPP currently pins scikit-image 0.26.0 in its supported Python 3.12 application environments.

Orthogonal 3D Skeleton QC¶

Use this small comparison before relying on a new volumetric segmentation:

Send the same explicit ZYX binary mask to two Skeletonize nodes. Set one to 3D ZYX with Lee and the other to 2D YX with Auto.
Inspect the source mask and both skeleton outputs in XY, XZ, and YZ views. A true 3D centerline should remain connected while it curves or branches through Z. A slice-wise result often appears plausible in XY but becomes a ribbon, a set of parallel paths, or disconnected fragments in XZ/YZ.
Connect each result to Skeleton Keypoints or Skeleton Graph Overlay and compare endpoints and junctions in all three orientations.
Connect each result to Analyze Skeleton. Review component, endpoint, junction, branch, and cycle counts. Large changes between the two modes are evidence that the spatial-processing choice materially affects network topology.
Keep 3D ZYX only when Z is an explicitly declared spatial axis. For an anisotropic volume, set the Z/Y/X calibration before interpreting physical branch lengths.

Analyze Skeleton¶

Input: skeleton mask, or a binary mask if Input is set to Skeletonize first.
Output: table, one row per connected skeleton component.
Purpose: Measures whole-network/component properties.
Reports: skeleton voxel count, endpoint voxels, junction voxels, isolated nodes, branch count, graph node/edge counts, voxel-graph edge count, cycle count, component context, and skeleton length in pixel/voxel units plus physical units when scale metadata is available.
Execution: manual/cached. Use Calculate or enable Auto Recalculate for small data.

Measure Skeleton Branches¶

Input: skeleton mask, or a binary mask if Input is set to Skeletonize first.
Output: table, one row per traced graph branch.
Purpose: Measures individual branches between graph nodes.
Reports: component ID, branch ID, branch type, voxel count, graph-edge count, branch length, endpoint-to-endpoint distance, tortuosity, start/end coordinates, and calibrated physical length when scale metadata is available.
Execution: manual/cached. This can produce many rows on dense networks.

Summarize Skeleton Branches¶

Input: a table from Measure Skeleton Branches.
Output: table, one row per grouping block. Auto grouping preserves useful context columns such as source, time, channel, and spatial block indices.
Purpose: Converts row-per-branch measurements into compact distributions suitable for treatment comparison, PCA-style feature extraction, and network QC.
Reports: branch count, component count, total branch length, selectable length and tortuosity statistics, and branch-type counts/fractions such as endpoint-to-junction or junction-to-junction fractions.
Use when: you want table-derived branch distributions without manually configuring the generic Summarize Measurements node.

Skeleton Graph Tables¶

Input: skeleton mask, or a binary mask if Input is set to Skeletonize first.
Output: two table outputs: graph nodes and graph edges.
Purpose: Exports the explicit graph representation for downstream network analysis, plotting, or external graph tools.
Node table reports: component ID, graph node ID, node type, node degree, skeleton voxel index, and spatial coordinates.
Edge table reports: component ID, edge ID, start/end node IDs, branch type, branch voxel count, branch edge count, path length, endpoint distance, tortuosity, start/end coordinates, and calibrated physical length when scale metadata is available.
Execution: manual/cached. This is the most direct table export of the skeleton graph.

Measure Overall Skeleton Network¶

Input: skeleton mask, or a binary mask if Input is set to Skeletonize first.
Output: table, one row per analyzed spatial block, such as one image, timepoint, channel, or slice depending on spatial processing.
Purpose: Measures whole-network topology directly from the skeleton mask. This is not just a table summary of Measure Skeleton Branches; it computes component-level and graph-level metrics that require the original skeleton graph.
Reports: component count, skeleton voxel count, largest-component fraction, isolated component count, endpoint/junction/isolated-node counts, branch and graph-edge counts, cycle count, total skeleton length, branch length summaries, mean tortuosity, connectedness fraction, and fragmentation index. It also reports normalized connectedness features such as isolated component fraction, branches/endpoints/junctions/cycles per component, and components/branches/endpoints/junctions/cycles per skeleton length or calibrated physical length when available.
Use when: you want one compact feature row per image, timepoint, channel, or other spatial block for treatment comparison, PCA-style analysis, or mitochondrial connectedness QC.
Execution: manual/cached.

Use Measure Skeleton Branches -> Summarize Skeleton Branches when you want branch-length distributions and branch-type fractions grouped by image, timepoint, channel, condition, or branch type. Use the generic Summarize Measurements node only when you need custom statistics on arbitrary table columns. Use Measure Overall Skeleton Network when you need network-level quantities such as connected components, cycles, isolated nodes, largest-component fraction, and normalized connectedness metrics.

Skeleton Keypoints¶

Input: skeleton mask.
Output: three mask outputs: endpoints, junctions, and isolated nodes.
Purpose: Visual QC of graph topology.
Use when: you need to check whether a segmentation/skeletonization step is creating too many breaks, junctions, or isolated fragments.

Skeleton Graph Overlay¶

Input: skeleton mask.
Output: channel-last RGB image.
Purpose: Visual QC overlay for graph topology in napari.
Display modes: colored edges with colored nodes, colored edges only, or white edges with colored nodes.
Node colors: endpoints are green, junctions are magenta, and isolated nodes are cyan/blue.
Use when: you want graph branches and nodes to be visually obvious. 2D overlays display as one RGB image layer; 3D overlays display as separate additive red/green/blue layers so napari can render the colors reliably.

Label Skeleton Components¶

Input: skeleton mask.
Output: label image.
Purpose: Assigns a label ID to each connected skeleton component.
Use when: you need to inspect or count disconnected skeleton networks.

Label Skeleton Branches¶

Input: skeleton mask.
Output: label image.
Purpose: Assigns label IDs to branch paths between graph nodes.
Use when: you need an inspectable branch map rather than a table.
Note: Junction voxels are deliberately not assigned to branch labels so connected branch paths remain visually separable.

Prune Skeleton Branches¶

Input: skeleton mask.
Output: binary skeleton mask.
Purpose: Removes short terminal spurs and optional isolated skeleton voxels.
Key settings: minimum terminal branch length, length units (Pixels/voxels or Physical units), pruning passes, isolated-voxel removal, and 2D/3D spatial processing.
Use when: thresholding or skeletonization creates small terminal artifacts that inflate endpoint and branch counts.
Physical units: require pixel-size/axis-scale metadata. If no calibration exists, physical-unit pruning behaves like unit spacing.

Interpreting Graph Terms¶

Endpoint: skeleton voxel/pixel with one graph neighbor.
Junction: skeleton voxel/pixel with three or more graph neighbors.
Isolated node: foreground skeleton voxel/pixel with no graph neighbors.
Branch: path between two graph nodes, usually endpoint-to-junction, junction-to-junction, endpoint-to-endpoint, or a cycle.
Tortuosity: branch path length divided by endpoint-to-endpoint distance. A straight branch has tortuosity near 1.

Common Pitfalls¶

Do not feed a thick binary mask into branch-label or keypoint nodes unless you intentionally want graph analysis on the thick mask. Usually run Skeletonize first.
For anisotropic 3D data, set pixel size / units before measurement so physical length columns use the correct z spacing.
For slice-wise analysis of a stack, explicitly use 2D spatial processing. For true volumetric connectedness, use 3D spatial processing.
If the graph has many tiny branches, inspect the segmentation first, then try Fill Holes, Remove Small Objects, or Prune Skeleton Branches.