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Spots

The Spots tab provides spot detection and colocalization visualization for analyzing fluorescent spots in your images.

Interface overview

Spot detection section

Controls:

  • Image layer (dropdown): Select the image layer containing spots to detect.
  • Detector (dropdown): Choose a spot detection algorithm.
  • Settings (dynamic panel): Detector-specific parameters.
  • Minimum diameter (spin box): Minimum spot diameter in pixels (0 = no minimum filter).
  • Maximum diameter (spin box): Maximum spot diameter in pixels (0 = no maximum filter).
  • Run (button): Execute spot detection on the selected image.

Diameter filtering behavior: After detection, connected components are filtered against diameter-derived thresholds:

  • For 2D labels, thresholds are converted to effective area: A = pi * (d / 2)^2.
  • For 3D labels, thresholds are converted to effective volume: V = (4/3) * pi * (d / 2)^3.

Components with pixel/voxel counts outside the computed range are removed.

Colocalization section

Controls:

  • Labels A (dropdown): First labels layer.
  • Labels B (dropdown): Second labels layer.
  • Visualize (button): Compute intersection and visualize overlaps.

Output: Creates a Points layer named {labels_A}_{labels_B}_colocalization with yellow ring markers at overlap locations.

Available detectors

Detector Algorithm Description
rmp Rotational Morphological Processing (RMP) Spot extraction with rotating images and thin structuring elements. Compatible with 2D and 3D images.
ufish U-FISH Spot extraction with a compact deep learning model for 2D and 3D images.

Output layers

  • Spot detection outputs <image layer>_<detector>_spot_labels.

Detector settings

rmp

Source: src/senoquant/tabs/spots/models/rmp/details.json and src/senoquant/tabs/spots/models/rmp/model.py.

Method reference: Rotational Morphological Processing for spot detection.

Setting Type Default Range Description
Spot diameter (px) int 10 3 - 9999 Expected spot diameter used by the extraction structuring element.
Auto threshold bool true n/a Uses Otsu thresholding on the normalized response.
Manual threshold float 0.50 0.0 - 1.0 Fixed threshold when Auto threshold is off. Disabled when auto-thresholding is enabled.

RMP simplifications: - Angle spacing is fixed internally to 5 degrees. - Wavelet denoising is always enabled (both before and after top-hat extraction).

ufish

Source: src/senoquant/tabs/spots/models/ufish/details.json and src/senoquant/tabs/spots/models/ufish/model.py.

Setting Type Default Range Description
Spot size float 1.0 0.25 - 4.0 Spot-scale control. 1.0 is default, >1 biases detection toward larger spots, <1 toward smaller spots.
Threshold float 0.5 0.0 - 1.0 Foreground threshold on the enhanced response (lower = more spots).

UFISH simplification: - Wavelet denoising is always enabled before enhancement/segmentation.