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Detection Engine

Tissue Detection

Finding where the tissue is on a slide

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Definition
Before you analyze cells, you need to know where the tissue is. Tissue Detection works on the preview image, the top level of the sample's cache, and separates tissue from background with an intensity threshold. It joins nearby fragments, drops tiny ones and inflates the outline slightly. The engine returns a white-on-black tissue mask, and the ROI tool of the same name turns the detection into regions.
H&EDSPTCH
Video · Primary
User Interface - Image Viewer + Toolbar
Video · Primary
Supporting
User Interface - Object Inspector
Video · Supporting
User Interface - How to use ROI, ANN and EA tool
Video · Supporting
TQ to SQ workflow
Video · Supporting
User Interface - Raw Data
Video · Supporting
Analysis Boundary
A mask later engines can use
Preview-Based Speed
Runs on the cache's top level
Automatic or typed threshold
Intensity Threshold, computed or entered
TMA Support
A separate TMA Detection tool

How It Works

Two things in StrataQuest share this name. The Tissue Detection engine sits in a layer's Pre-Processing section (Add > Detection > Tissue Detection) and returns a mask, tissue white and everything else black. The Tissue Detection tool, opened with Autodetect ROIs on the ROI toolbar, uses the same settings to draw regions of interestLoading... on the sample. Both run on the preview image, the top level of the cache, so the cache must be built first.

The settings describe four adjustments. An intensity threshold, computed automatically or typed, separates tissue from background. Fragments within Tissue Fragments – max. merging distance are joined into one region, regions smaller than Tissue Fragments – min. area are dropped, and Contour Inflate Size widens each region by a number of pixels so the tissue edges come along. Exclude border connected regions drops regions touching the border.

The Engines Reference's recipe is to start permissive and tighten. Set the threshold low so everything shows, raise it until the tissue is right, raise the merging setting in small steps until fragments join, and raise the minimum area last to clear small specks. Detection Profiles save a tuned set of values for reuse.

Simplified

The engine works on a small preview of the whole slide and classifies each pixel as tissue or not. Then it joins nearby pieces, drops specks and widens the outline a little so the tissue edge stays in.

Science Behind It

The image pyramid. A whole-slide scan holds far more pixels than the tissue outline needs. In digital pathology generally, scans are stored as pyramids: the same image at several resolutions, each level a fraction of the one below. Finding the edge between tissue and glass needs only a coarse level, which makes the job fast. StrataQuest's cache is this kind of pyramid, and Tissue Detection reads its top level.

Thresholds and histograms. On many brightfield previews the histogramLoading... of pixel values shows two populations, bright glass and darker tissue, with a valley between them, and a method such as Otsu'sLoading... picks a cutoff in that valley. The Engines Reference doesn't name the method behind Automatic, though it does name Otsu for the Nuclei and Total Area engines. Its own example is a fluorescence sample, where tissue is typically brighter than the background, so don't assume tissue is always the darker class.

From mask to regions. After thresholding, the tissue is a set of separate patches. In image processing generally, connected component labelingLoading... turns patches into distinct regions, each with its own number. Merging distance and minimum area then act on regions: the first joins pieces of one section split by tears or faint staining, and the second drops dust and debris.

Why inflate. The outermost cells of a section are often faint, or only partly inside a tight outline. Contour Inflate Size adds a margin of pixels so the tissue edges and a small surrounding area are included. A layer's Events Strategy then determines what happens to cells cut by a region's boundary.

Simplified

Whole-slide images are huge, so the engine reads a small preview instead. On a preview, glass and tissue usually differ clearly in brightness, and a single cutoff separates them. The outline is then widened a little so the cells at the tissue edge stay in.

Parameters & Settings

ParameterDescription
Input imageThe image for tissue detection. Once it is set, the Tissue Detection button opens the Tissue Detection window.
AutomaticWhen checked, the Intensity Threshold is computed automatically on the gray image within the range you select, and shown in the edit box.
Intensity ThresholdThe intensity threshold. Uncheck Automatic to type a value; the Gray indicator shows pixel intensities in the gray image.
Tissue Fragments – max. merging distanceIncrease it if a region is split into many small subregions; the result is a single region containing them.
Tissue Fragments – min. areaThe smallest area a tissue region can have; smaller regions are not considered.
Contour Inflate SizeThe detected region is inflated by this number of pixels, to include the tissue edges and a small surrounding area.
Exclude border connected regionsRegions touching the border are excluded from detection.
Maximum number of regionsROI tool only (Autodetect ROIs): the maximum number of regions to detect.
Detection ProfilesDefault stores the current values; Restore Defaults returns to the last saved defaults; Profile saves the settings; Load Profile loads a saved profile.

In the ROI tool the User Manual describes Automatic differently: it determines red, green and blue channel values within three intervals, from a lower value (default 5) to an upper value (default 255). Applying Illumination Correction first improves Autodetect ROIs.

Source: StrataQuest 8.1 Engines Reference, ch. 44 (pp. 550–553); User Manual §10.3.1–10.3.2 (pp. 91–98), §11.1.1 (pp. 155–157).

Simplified

Leave Automatic on to start. If tissue comes out in pieces, raise the merging distance; if specks of dust appear, raise the minimum area; if edge cells are left out, raise Contour Inflate Size.

Practical Example

Suppose you have a tissue microarray slide with many cores, and a whole-section slide from the same study.

  1. Build the cache for each sample, and apply Illumination Correction.
  2. Whole section: press Autodetect ROIs. In the dialog, start with a low Intensity Threshold, raise it until the section is outlined, then raise Tissue Fragments – max. merging distance until the section is one region. Save the settings as a profile for the rest of the batch.
  3. Microarray: press TMA Detection on the toolbar instead. If spots are broken or the contrast is poor, set Spot Mean Radius by hand, then choose Create Samples for each Spot.
  4. Inside a layer: add the Tissue Detection engine in Pre-Processing when a later engine needs a tissue mask as input.

Each TMA spot becomes its own sample, so NucleiLoading... detection, measurements and diagrams are reported per core.

Simplified

For a whole section, Autodetect ROIs outlines the tissue for you. For a microarray, the TMA Detection tool finds each core and can make it its own sample, so every core gets its own counts and statistics.

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