phenix-icechunk

Expose an Opera Phenix / Harmony Index.xml export as a virtual OME-Zarr 0.5 HCS plate inside an icechunk repo. No pixels are copied: every zarr chunk is a virtual reference into the original TIFFs, so the repo is a few hundred KB.

Setup

uv venv --python 3.12 .venv
uv pip install --python .venv/bin/python -e .

Convert

.venv/bin/phenix-icechunk "/path/to/Measurement 2/Images/Index.xml" data/plate.icechunk

# + flatfield correction by default, + Bio-Formats OME-TIFFs (one per well) next to the repo
.venv/bin/phenix-icechunk "/path/to/Measurement 2/Images/Index.xml" data/plate.icechunk \
    --flatfield --ome-tiff data/ome_tiff

# + max z projection (after flatfield), committed as its own transaction, exported as OME-TIFF
.venv/bin/phenix-icechunk "/path/to/Measurement 2/Images/Index.xml" data/plate.icechunk \
    --flatfield --zproject --ome-tiff data/ome_tiff        # --zproject mean|median|min|sum|std

Read as xarray

import zarr
from phenix_icechunk import open_plate, open_field, open_datatree

zarr.config.set({"async.concurrency": 16})        # default 10; 16 was best on /nfs
plate = open_plate("data/plate.icechunk")         # open once, reuse for many fields

da = open_field(plate, "A4", 0)                   # lazy xr.DataArray, nothing read yet
dapi = da.sel(c="DAPI").isel(t=0).values          # reads only the 12 DAPI planes
roi = da.sel(x=slice(1900, 2000), y=slice(-7100, -7000))  # select in µm
da.attrs["plate_id"], da.attrs["assay_layout"], da.coords["acquisition_time"]

da = open_field(plate, "A4", 0, chunks={})        # same, dask-backed (needs dask)

tree = open_datatree(plate)                       # whole plate as xr.DataTree, all lazy
tree["A/4/17"]["image"]                           # field 17 of well A4
tree["phenix/images"]                             # per-image table as a Dataset

open_field returns dims (t, c, z, y, x) with:

• coords:
  • c: channel names, plus channel_id, excitation/emission wavelength, exposure time, acquisition type, binning, camera, objective, colour.
  • z: plane z in µm, plus plane_id.
  • y, x: µm in stage space. This is the same transform as the OME-Zarr metadata, so neighbouring fields line up.
  • t: seconds, plus timepoint_id.
  • Per-plane (t, c, z) coords: image_id, acquisition_time (datetime64, UTC), abs_position_z, intensity_mean/stddev/min/max, temperature, co2, humidity. Pick other table columns with plane_coords={"name": "column", ...}.
• attrs: well (ID, row/col, assay layout), field (ID, stage position, acquisition start/end, OME-XML image ID), plate (IDs, type, measurement, user, instrument, source Index.xml), and the field's full OME-Zarr metadata under attrs["ome"].

Reads through xarray cost the same as raw zarr: about 0.35 s for a full 4×12-plane field. Nothing is decoded to float, because fill values are not applied, so pixels stay uint16.

Flatfield correction

from phenix_icechunk import flatfield_profiles

da = open_field(plate, "A4", 0, flatfield=True)   # still lazy, float32
# flatfield=None (default) follows the repo: True if it was converted with --flatfield
da.attrs["flatfield"]                             # formula + per-channel profile summary
tree = open_datatree(plate, flatfield=True)       # same for every field
flatfield_profiles(plate)                         # the (c, y, x) profiles themselves

Each channel's Harmony FlatfieldProfile (Background, an Acapella 2-D polynomial) is evaluated as P = Σ Coefficients[k][j] · u^(k−j) · v^j, with u = (x − Origin₀)·Scale₀ and v = (y − Origin₁)·Scale₁. It is normalised to mean 1, and pixels become raw / P, so intensities keep their scale.

• The correction is applied on read, per plane. Nothing in the repo changes, and it costs about 0–0.1 s per full field on top of the 0.5 s read.
• The term order was checked against the data. The median of all 56 fields per channel correlates with P at 0.95–0.98, against 0.52–0.81 for the transposed order. After correction, background non-flatness drops from 3.6–6.3 % to 0.9–2.1 %, with the mean unchanged.
• Channels without a polynomial (e.g. Character: Flat) are left as they are (attrs["flatfield"]["channels"][name]["applied"] == False).
• If the image size differs from the profile's Dims (binning), the profile is rescaled.

Z projection

from phenix_icechunk import zproject

zproject("data/plate.icechunk", "mean")            # add another method later: one more commit
mip = open_field(plate, "A4", 0, zproject="max")   # (t, c, 1, y, x), attrs["zproject"]
tree = open_datatree(plate, zproject="max")

• Projections are real pixels (they can't be virtual). They are stored as a full OME-Zarr HCS plate under /projections/z<method>/<row>/<col>/<field>/0 (z size 1, z = mid-stack), and listed in root attrs["phenix"]["projections"].
• Each projection is its own icechunk commit on top of the virtual plate, e.g. Z max projection of 56 fields after flatfield correction. The plate commit is never touched, so a failed or replaced projection leaves it intact. Re-running a method needs overwrite=True, and the old version stays in history.
• Order: project(flatfield(raw)). The stored projection already has the correction, so open_field(..., zproject=...) refuses a conflicting flatfield= argument.
• dtype: max/min of raw planes stay uint16. With flatfield, or for mean/median/sum/std, the result is float32. For this plate the float32 max projection takes 3.3 GB in the repo and took 40 s to compute.

OME-TIFF export (Bio-Formats / Fiji / QuPath / OMERO)

--ome-tiff DIR on convert, or later from any repo:

from phenix_icechunk import export_ome_tiff
export_ome_tiff("data/plate.icechunk", "data/ome_tiff", flatfield=True, compression="zlib")
export_ome_tiff("data/plate.icechunk", "data/ome_tiff", zproject="max")  # -> <plate>_<well>_zmax.ome.tiff

The output is one <plate>_<well>.ome.tiff per well:

• Every field of view is its own series (tile): XYZCT planes, 512×512 internal tiles, BigTIFF.
• The OME-XML in the first IFD carries the Plate/Well/WellSample layout with stage positions, channels (names, wavelengths, modes, colours), every Plane (ΔT, exposure, XYZ), StageLabel, ImagingEnvironment, instrument, and assay-layout/channel MapAnnotations. It is the same document as /OME/METADATA.ome.xml, restricted to the well, plus TiffData per image.
• With flatfield, corrected pixels are rounded and clipped back to uint16.

Checked with Bio-Formats 8.1.1 showinf: the file opens as OME-TIFF with 56 series of Z=12 × C=4 × 2160², and the plate/well/well-sample positions are parsed. Planes extracted with bfconvert are bit-identical to the corrected repo planes. The well above (25 GB raw) took about 100 s to write and is 10.5 GB with zlib.

Low-level (zarr)

img = plate["A/4/0/0"]                           # zarr.Array, (t, c, z, y, x)
stack = img[0]                                   # numpy

open_plate grants icechunk access to the repo's own file:// container. Opening the repo with plain icechunk.Repository.open also works, but you have to pass authorize_virtual_chunk_access yourself.

from phenix_icechunk import read_file, read_table

df = read_table(plate["phenix/images"])                 # one row per <Image>, 76 columns
xml = read_file(plate["phenix/files/Images/Index.xml"]) # original bytes
ome = read_file(plate["OME/METADATA.ome.xml"]).decode() # generated OME-XML
plate["phenix"].attrs["index_xml"]["channels"]["1"]     # every Map entry for channel 1

What is stored

Every piece of metadata in the export is stored in the repo, both verbatim and parsed:

The plate, well and field metadata validate with ome-zarr-models (v0.5). Only the pixel strip offsets are not stored as metadata, because they are the virtual chunk refs.

How it works

• Index.xml gives the well/field/plane/channel/timepoint of every TIFF. Each field becomes one (t, c, z, y, x) array with OME-Zarr multiscales (scale from ImageResolution, z step from PositionZ, translation from stage position: x = +stageX, y = −stageY).
• TIFF headers are scanned in parallel (about 3 ms per file on /nfs) to get strip offsets and file sizes.
• Harmony writes two TIFF layouts, and both are supported:
  • LZW strips, e.g. 18 strips of 122 rows (OT_NIM).
  • LZW 512×512 tiles in newer exports, e.g. 25 tiles per 2160² plane (large_GBM).
• --chunking auto (default) picks tile for tiled exports and plane for strip exports.
• plane chunking: one chunk per plane, referencing the whole TIFF file and decoded by imagecodecs' imagecodecs_tiff codec (libtiff). That is one large read per plane, which is what NFS serves fastest, and it works the same for strips and tiles.
• strip / tile chunking: one chunk per TIFF strip or tile, whichever the files use, decoded by the bundled phenix_tiff_strip codec. The short last strip is zero-padded by the codec, and padded edge tiles are clipped by zarr. Partial reads touch fewer bytes, but the request count goes up 18–25×.
• Uncompressed TIFFs with contiguous strips get a plain bytes-codec chunk per plane.
• Each reference stores the TIFF's mtime, so icechunk raises an error rather than return wrong pixels if a source file is rewritten.

The codecs are registered through the zarr.codecs entry point, so any zarr-python 3 reader in this environment can decode the chunks. Readers elsewhere need imagecodecs (plane mode) or this package (strip mode).

Benchmark (bench/bench_read.py)

The measurement has 56 fields × 4 channels × 12 z planes of 2160² uint16, LZW-compressed. The table shows one field (448 MB decoded); each method read a different field from a cold cache.

Strip chunking does not pay off on this NFS mount, because per-request latency dominates. It may help on object storage or for viewers that fetch small tiles.

Notes / limitations

• icechunk 2.3 cannot match a virtual-chunk container whose prefix needs percent-encoding (for example the space in Measurement 2), so the container is set at the deepest URL-safe parent directory.
• Only one resolution level is exposed; pyramids would have to be materialised.
• Flatfield correction is read-time only (flatfield=True) and uses the Background profile. No darkfield/offset is subtracted, because Harmony exports here carry no separate darkfield, and the Background profile already describes the raw background including the camera offset.
• icechunk only accepts zarr keys, so files are stored as uint8 arrays, not plain files. Tools expecting bioformats2raw's plain OME/METADATA.ome.xml file need read_file.
• All channels must share image size, dtype and TIFF layout. Otherwise conversion stops with an error.