Parallax Pathology · Technical note
Russell Parrish · analysis carried out with Claude (Anthropic) · Version 2, 4 November 2026
TCGA slides are distributed by the GDC as Aperio SVS files and by the IDC as DICOM whole-slide images [1, 2]. Most pipelines read pixel spacing from file metadata to tile slides at a fixed physical scale. A file without a spacing tag is usually dropped or assigned a default; a file with a wrong tag is tiled at the wrong physical scale without any warning. Site-related differences in TCGA slide images are well documented [3]; this note concerns one specific, checkable case.
Header census. DX slides were listed through the GDC API. TIFF headers were read by HTTP byte ranges (about 1 GB of header bytes in total): all 459 TCGA-COAD DX slides, and for TCGA-READ every DX file under 20 MB plus two per tissue source site (41 slides). For each slide we recorded dimensions, pyramid levels, compression and the Aperio ImageDescription tags (MPP, AppMag, scanner, date).
IDC. DICOM series for TCGA sites AA and AG were queried with idc-index 0.13.4 (IDC v25, the latest index it provides at the time of writing), recording PixelSpacing and TotalPixelMatrix dimensions.
Scale bearings (17 COAD derivatives).
Downstream sets. File listings of Zenodo 3784345 [4] and the ZIP central directories of the CRC_DX archives of Zenodo 2530835 [5], read by HTTP range. Tile coordinates were read from locally held archives of Zenodo 3784345.
No clinical or outcome data were used.
| Route | DX slides | Sites | Scale tags |
|---|---|---|---|
| Aperio-native scan | 283 | all except AA, T9 | 40×, 0.245–0.253 µm/px |
| MIRAX scan converted to SVS | 159 | all 156 regular AA, 3 T9 | 20×, 0.2325 µm/px |
| Small derivative | 17 | AA only | none (no MPP, magnification, scanner or date) |
TCGA-READ adds 8 small derivatives at site AG (Indivumed's rectal code), for 25 in total; all 8 lack MPP and magnification tags. The 17 COAD derivatives are written by Aperio Image Library v12.1.3. Frozen-section (TS and BS) slides exist for all 25 patients (51 for the COAD patients, 20 for the READ patients); the COAD ones are full-size scans tagged 0.5015 µm/px. GDC records a creation date of 2021-10-13 for 351 of the 459 COAD DX files across all three routes (the other 108 are dated 2018-05-21). GDC Data Release 31.0 (29 October 2021) states that "All TCGA slide images that were removed earlier this year have been restored" [6]; the shared date is consistent with that restoration and carries no information about the derivatives' origin. No GDC release note we read mentions these files.
Each of the 25 derivatives appears in IDC v25 with the same pixel matrix as the GDC file and PixelSpacing 0.0005 mm. No other AA or AG DX series in IDC carries this value; the 228 others carry 0.00023 mm. The source SVS files have no spacing tag, so the 0.5 value was assigned during or after conversion.
| Bearing | Median µm/px | Per-slide IQR | Lowest slide | Main assumption |
|---|---|---|---|---|
| Scanner-field grid | 0.90 | 0.82–0.98 | 0.62 | same camera-field size as the regular Indivumed scans |
| Tissue area | 1.20 | 1.14–1.40 | 0.90 | derivative sections are of typical size |
| Structure scale | 1.26 | 1.09–1.49 | 0.54 | compression and blur do not shift the calibrated curve |
| Zenodo 3784345 tile grid | 1.28 | (one value) | an implied choice by the tile authors, not a measurement |
The bearings differ from each other by up to about 40%, but their medians all lie between 0.9 and 1.3 µm/px, 1.8 to 2.6 times coarser than the stated 0.5, and every interquartile range excludes 0.5. Per-slide values are noisy: single slides reach 0.62 (field grid) and 0.54 (structure scale), and no bearing resolves the scale of an individual slide. The field-grid bearing is the most direct, a physical ruler present in each image, and gives the finest value; the tissue-area bearing would read too coarse if these sections are smaller than typical. A spacing of 0.93 µm/px would equal a 4-times reduction of the 0.2325 µm/px MIRAX scans; this is consistent with the field-grid median but not established.
Zenodo 3784345 (598 patients; "512x512 px at 0.5 µm/px" per its record [4]) contains all 25 derivative patients; two of their archives (TCGA-AA-3488, TCGA-AG-3732) are 1,008 bytes and hold no tiles. For the 15 COAD derivative patients with non-empty archives held locally, every tile origin lies within the derivative's 10,000-px frame and the tile side is 200 px, so the tiles were cut from these files and enlarged to 512 px. At 0.9–1.3 µm/px a 200-px crop spans 180–260 µm against the nominal 256 µm, giving about 0.35–0.51 µm per tile pixel instead of 0.5, and no detail finer than the source pixel. Zenodo 2530835 (360 CRC_DX patients) contains none of the 25.
In exploratory work on the Zenodo 3784345 tiles, the derivative patients separated from all others on simple image measures and on a tissue classifier. The figures below are from the authors' own analysis and are given as an example of how the files behave, not as a general estimate.
| Tiles from | Laplacian variance (median) | Classifier: tumour | Fat | Background | Mucus |
|---|---|---|---|---|---|
| Aperio-native sites | 1,099 | all non-derivative patients: 0.577, 0.008, 0.007, 0.033 | |||
| Regular Indivumed scans | 215 | ||||
| Derivative patients | 18 | 0.351 | 0.130 | 0.119 | 0.162 |
Classifier: frozen DINOv2 ViT-S/14 features with a logistic probe trained on the NCT-CRC-HE-100K-NONORM tiles [7]; mean predicted share over quadrants of 30 tiles per patient.
Scripts, outputs and SHA-256 hashes are held by the authors (folders TCGA_PRODUCTION_ROUTES_20261104 and DERIVATIVE_NOTE_20261104) and are available on request. Only public metadata and image pixels were used.
| Patient | Project | GDC file ID | Pixels (w × h) | MB | Field-grid µm/px |
|---|---|---|---|---|---|
| TCGA-AA-3488 | COAD | b8085eff-8407-4fd5-b2ba-70fa649aa07d | 10000 × 7918 | 7.8 | 1.02 |
| TCGA-AA-3489 | COAD | 35c1e23c-8adb-4a7f-865f-be1d6e10b3ef | 8711 × 10000 | 7.3 | 0.97 |
| TCGA-AA-3492 | COAD | fa3a810e-09c6-4342-8923-1fe70546d9de | 9018 × 10000 | 8.6 | 0.98 |
| TCGA-AA-3494 | COAD | b6c0ea5f-32d3-4210-b85b-9cb435e8167e | 8087 × 10000 | 8.7 | 0.82 |
| TCGA-AA-3495 | COAD | daff3278-bdad-4db0-96c8-290918dd99ac | 8125 × 10000 | 10.0 | 0.70 |
| TCGA-AA-3496 | COAD | efd81ef1-0260-4e2c-ad3e-64733b3072a2 | 9734 × 10000 | 8.2 | 1.07 |
| TCGA-AA-3506 | COAD | 76c04cfc-4bf8-486e-ad34-c60da2d3d64b | 8447 × 10000 | 10.3 | 0.90 |
| TCGA-AA-3509 | COAD | 3ac10c45-4096-4839-982d-3019e77587db | 7948 × 10000 | 11.4 | 0.90 |
| TCGA-AA-3510 | COAD | 9b05cf34-c548-482f-9e41-a77e2ba5a253 | 8520 × 10000 | 9.5 | 0.62 |
| TCGA-AA-3511 | COAD | cdcd8072-7739-496e-ac05-6df73e3cd940 | 10000 × 7175 | 10.0 | 0.97 |
| TCGA-AA-3655 | COAD | 866108f6-0a38-4493-9eb5-864ae8b1eb4a | 10000 × 8593 | 12.8 | 0.89 |
| TCGA-AA-3660 | COAD | 1ca35a8e-3911-4247-a906-832f3223e192 | 10000 × 6751 | 8.5 | 0.86 |
| TCGA-AA-3662 | COAD | 6fccd4e0-9cc3-4abe-b216-e1049284a4a4 | 10000 × 7946 | 9.5 | 0.80 |
| TCGA-AA-3663 | COAD | 5fe3d9b5-c3f6-46d7-a06d-84e395017cf6 | 10000 × 9869 | 12.0 | 0.88 |
| TCGA-AA-3697 | COAD | 5f4f67bf-5b13-4aa4-929b-9aa096bb89d5 | 7081 × 10000 | 9.6 | 1.77 |
| TCGA-AA-3712 | COAD | 3962ba26-19f7-46be-af73-e39b37a51c64 | 6298 × 10000 | 8.2 | 0.74 |
| TCGA-AA-3713 | COAD | a72573a6-2462-4185-b4e8-a081c95d8fd7 | 7645 × 10000 | 8.2 | 1.28 |
| TCGA-AG-3591 | READ | 82c5f12b-1a92-4d33-8f5d-7a703bcd4ef9 | 10000 × 7799 | 7.6 | not measured |
| TCGA-AG-3592 | READ | 15a3d699-1c64-4711-969b-dbffda0a5f9f | 10000 × 6751 | 10.1 | not measured |
| TCGA-AG-3725 | READ | 2cd68fa7-8178-478f-bd58-aeaa0fe42380 | 10000 × 9089 | 4.7 | not measured |
| TCGA-AG-3731 | READ | ff868863-d558-4e5d-b0f7-0f1dc7b21fe8 | 6043 × 10000 | 9.1 | not measured |
| TCGA-AG-3732 | READ | 0f4229c7-5a59-41d4-aecf-50388c12336d | 8293 × 10000 | 7.7 | not measured |
| TCGA-AG-3742 | READ | 7d2955fd-c31d-4e26-93b0-a9e38195cf7e | 7186 × 10000 | 7.7 | not measured |
| TCGA-AG-3891 | READ | d024059f-5466-4fc7-a383-6a4bf428245c | 10000 × 9092 | 12.4 | not measured |
| TCGA-AG-4009 | READ | 67cd0eac-769d-45d6-be6f-1a852a8a897e | 9445 × 10000 | 8.8 | not measured |
Per-slide field-grid values are individually uncertain (Section 4); IDC states 0.5 for every row.