Research workbench

IHC DAB Analysis

Free online IHC DAB staining analyzer. Colour deconvolution, DAB quantification, H-Score calculation. No upload, runs in browser.

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Drop IHC image here or click to upload

Drop or click to add JPEG / PNG files · Multiple files supported · 20MB max each

PNGJPG

Current Method

Ruifrok Deconvolution

Best for hematoxylin + DAB slides because it separates counterstain from DAB signal.

Threshold Source

Auto · Otsu

Estimated from the current image and threshold method. Good as a starting point.

Scoring Mode

Relative

Splits weak / moderate / strong from this image’s DAB range. Best for single-image exploration.

White Reference

rgb(255, 255, 255)

If the background is yellowish or gray, calibrating white first improves OD and tissue detection.

⚙️ Advanced Settings ▸

What this tool measures right now

This browser-based analyzer is designed for hematoxylin + DAB images that have already been cropped to a representative region. It separates brown DAB signal from the blue counterstain, detects tissue pixels, and reports positive tissue percentage, mean DAB optical density, H-Score, and optional DAB intensity summaries such as IntDen and CTCF.

Ruifrok deconvolution for stain separation, plus an HSV fallback for quick brown-signal checks.

Threshold-based quantification of positive tissue within the detected tissue mask.

Pixel-based H-Score using relative, fixed-OD, or IHC Profiler-style grading modes.

Live overlay preview so you can see whether the thresholding agrees with the visible stain pattern.

Recommended workflow

Crop first. Use a representative field or ROI before uploading. This tool does not draw ROIs or navigate whole-slide images.

Start with Ruifrok. For standard hematoxylin + DAB slides, colour deconvolution is usually more stable than HSV.

Check the overlay. Make sure the highlighted pixels match the brown stain you intend to quantify.

Only then tune threshold or white reference. If the background is not neutral white, use the eyedropper before comparing results.

Export your settings with the result. Threshold and scoring mode directly affect the numbers.

Interpret with care

This is not cell segmentation. The reported H-Score is calculated from pixel intensities inside the detected tissue mask, not from counted cells or nuclei.

Comparison is only meaningful when acquisition is consistent. Keep microscope, illumination, exposure, and stain protocol as constant as possible.

Use fixed OD cutoffs for cross-image comparison. Relative mode is convenient for exploring one image, but fixed mode is better when you want consistency across a cohort.

Best suited to exploratory research workflows. This tool is not a clinical diagnostic system.

Frequently Asked Questions

Is any image data uploaded to a server?

No. The image stays in your browser. All stain separation and quantification are executed client-side.

Which stain combinations are supported?

The current implementation is tuned for hematoxylin + DAB. It does not expose a custom stain-matrix editor in the UI.

Can I use it on a full whole-slide image?

Not directly. Large slides should be cropped to a representative field before analysis so the preview remains interpretable and the browser stays responsive.

What does “positive tissue (%)” mean?

It is the fraction of pixels above the DAB threshold within detected tissue, not the fraction of the entire uploaded canvas.

IHC DAB Analysis

Analysis Settings Summary