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Microscopy TIFFs come in many flavors: 16-bit or float pixels, single or
multi channel, and multi-page Z / time stacks. ``standardize_image`` collapses
any of those variants to a single canonical 8-bit RGB PNG used for BOTH the
on-screen preview and the model, so what you see is what gets segmented.
Which axis means what is read from the file's own metadata (``series.axes``
from tifffile: 'C'=channel, 'S'=RGB samples, 'Z'/'T'=stack, 'Y'/'X'=spatial)
rather than guessed from the array shape -- guessing cannot tell a 3-channel
(C, Y, X) image apart from a 3-slice (Z, Y, X) stack. Only when a file names
no axes at all (tifffile reports 'Q' = unknown) do we fall back to the dominant
convention: the first unknown axis of size 2-4 is the channel axis.
Reduction policy:
* multi-page / Z / time stacks -> one frame (default: the first)
* more than 3 channels -> first 3 channels (as R, G, B)
* intensity -> 0-255 -> 1st-99th percentile auto-contrast for
16-bit/float input (outlier-robust: a hot /
saturated pixel would make plain min/max
scaling collapse the real signal to black)
Standard 8-bit inputs (PNG/JPG/8-bit TIFF) are passed through unchanged in
value; only channel layout is normalized.
"""
import os
import tempfile
from typing import NamedTuple
import numpy as np
from PIL import Image
try:
import tifffile
except ImportError: # pragma: no cover - tifffile is a project dependency
tifffile = None
# Size policy. The model resizes any input to 512x512 (see segmentation.py), so a
# larger image costs memory/time and *loses* detail rather than adding any.
RECOMMENDED_SIZE = 512 # cropping to about this preserves the most detail
WARN_SIZE = 3072 # above this, advise the user to crop
MAX_SIZE = 4096 # above this, refuse: reading the pixels risks an OOM
# Refuse files whose full array would not comfortably fit in memory. Measured on
# the uncompressed array (shape x dtype) rather than the file size on disk, since
# compression makes disk size a poor proxy for what we actually allocate.
MAX_READ_BYTES = 300 * 1024 ** 2 # 300 MiB
# Axis roles, per tifffile's `series.axes` naming.
_SPATIAL = ("Y", "X")
_CHANNEL = ("C", "S") # C = separate channel planes, S = interleaved RGB samples
_UNKNOWN = ("Q", "I") # file named no axis; only these may be *guessed* as channels
# Anything else (Z, T, ...) is a stack axis and is indexed by frame.
def _read_array(path):
"""Load an image file into (array, axes) where axes names each dimension.
axes uses tifffile's convention ('C' channel, 'S' RGB samples, 'Z'/'T'
stack, 'Y'/'X' spatial, 'Q' unknown). Indexed / palette images (ImageJ
"8-bit Color", palette PNG/GIF) are expanded through their color lookup
table so we return true RGB, not bare indices.
"""
ext = os.path.splitext(path)[1].lower()
if ext in (".tif", ".tiff") and tifffile is not None:
with tifffile.TiffFile(path) as tif:
series = tif.series[0]
page = tif.pages[0]
arr = np.asarray(series.asarray())
axes = str(series.axes)
is_palette = getattr(page, "photometric", None) == tifffile.PHOTOMETRIC.PALETTE
if is_palette and page.colormap is not None:
colormap = np.asarray(page.colormap) # (3, 2**bits), uint16
rgb = np.moveaxis(colormap[:, arr], 0, -1) # (..., H, W, 3)
# TIFF colormaps are 16-bit; scale down to 8-bit (65535/255=257).
arr = np.round(rgb / 257.0).astype(np.uint8)
axes = axes + "S" # the LUT added an RGB sample axis
return arr, axes
# Everything else (and if tifffile is unavailable): PIL. Expand palette
# images to RGB so the LUT is applied instead of returning bare indices.
img = Image.open(path)
if img.mode in ("P", "PA"):
img = img.convert("RGB")
arr = np.asarray(img)
return arr, ("YXS" if arr.ndim == 3 else "YX")
def _shape_axes(path):
"""Return (shape, axes) from metadata only - no pixel decode."""
ext = os.path.splitext(path)[1].lower()
if ext in (".tif", ".tiff") and tifffile is not None:
try:
with tifffile.TiffFile(path) as tif:
series = tif.series[0]
return tuple(series.shape), str(series.axes)
except Exception: # noqa: BLE001 - fall through to PIL
pass
with Image.open(path) as img:
w, h = img.size
if img.mode in ("P", "PA", "RGB", "RGBA", "CMYK", "YCbCr", "LAB", "HSV"):
return (h, w, len(img.getbands())), "YXS"
return (h, w), "YX"
def _plan_axes(shape, axes):
"""Drop size-1 axes, infer a channel axis when the file names none, and move
it last.
Returns (shape, axes, guessed) as lists + a flag saying whether the channel
axis had to be inferred rather than read from the file.
"""
axes = list(axes)
shape = list(shape)
if len(axes) != len(shape): # be defensive; keep the trailing spatial axes
axes = ["Q"] * (len(shape) - 2) + ["Y", "X"]
pairs = [(a, d) for a, d in zip(axes, shape) if d != 1]
axes = [a for a, _ in pairs]
shape = [d for _, d in pairs]
guessed = False
if not any(a in _CHANNEL for a in axes):
# Only guess on axes the file left unnamed. An axis the file explicitly
# calls Z/T is a stack even when its length happens to be 3.
for i, (a, d) in enumerate(zip(axes, shape)):
if a in _UNKNOWN and d in (2, 3, 4):
axes[i] = "C"
guessed = True
break
ci = next((i for i, a in enumerate(axes) if a in _CHANNEL), None)
if ci is not None:
axes.append(axes.pop(ci))
shape.append(shape.pop(ci))
return shape, axes, guessed
def _reduce_to_hwc(arr, axes, frame=0):
"""Collapse a labelled array to 2D (H, W) or 3D (H, W, C).
The channel axis (named by the file, or inferred by ``_plan_axes`` only when
the file names none) is moved last and kept whole. The remaining stack axes
(Z / time / page) are indexed: the first by ``frame``, any deeper ones by 0.
"""
axes = list(axes)
# Drop size-1 axes, keeping arr and axes in sync.
for i in range(len(axes) - 1, -1, -1):
if arr.shape[i] == 1:
arr = arr.reshape(arr.shape[:i] + arr.shape[i + 1:])
axes.pop(i)
_, planned, _ = _plan_axes(arr.shape, axes)
# Apply the plan to the array: infer the channel axis, then move it last.
if "C" in planned and not any(a in _CHANNEL for a in axes):
for i, (a, d) in enumerate(zip(axes, arr.shape)):
if a in _UNKNOWN and d in (2, 3, 4):
axes[i] = "C"
break
ci = next((i for i, a in enumerate(axes) if a in _CHANNEL), None)
if ci is not None:
arr = np.moveaxis(arr, ci, -1)
axes.append(axes.pop(ci))
# Index the leading stack axes: first by `frame`, deeper by 0.
target = 3 if ci is not None else 2
first = True
while len(axes) > target:
idx = int(frame) if first else 0
idx = max(0, min(idx, arr.shape[0] - 1))
arr = arr[idx]
axes.pop(0)
first = False
return arr
class ImageInfo(NamedTuple):
"""How a file's dimensions were interpreted (read from metadata only).
frames - length of the first stack (Z/T) axis, 1 if not a stack
channels - length of the channel axis, 1 if single-channel
axes - the file's own axes string, e.g. 'CZYX' ('Q' = unnamed)
guessed - True if the channel axis was inferred rather than read
shape - the file's raw shape as stored, e.g. (1, 4, 1, 1024, 1024)
width - pixels along the X axis (0 if unknown)
height - pixels along the Y axis (0 if unknown)
"""
frames: int
channels: int
axes: str
guessed: bool
shape: tuple
width: int
height: int
def inspect_image(path):
"""Describe a file's structure from metadata only (no pixel decode)."""
try:
shape, axes = _shape_axes(path)
planned_shape, planned_axes, guessed = _plan_axes(shape, axes)
has_c = bool(planned_axes) and planned_axes[-1] in _CHANNEL
channels = int(planned_shape[-1]) if has_c else 1
target = 3 if has_c else 2
frames = int(planned_shape[0]) if len(planned_axes) > target else 1
# Spatial size comes from the named Y/X axes, so it stays correct
# whatever order the other axes are in.
width = height = 0
for a, d in zip(axes, shape):
if a == "Y":
height = int(d)
elif a == "X":
width = int(d)
if not (width and height): # no Y/X named; fall back to the header probe
width, height = image_size(path)
return ImageInfo(frames, channels, str(axes), guessed, tuple(shape), width, height)
except Exception: # noqa: BLE001
return ImageInfo(1, 1, "", False, (), 0, 0)
def count_frames(path):
"""Return how many frames a file's stack (Z/T) axis has (1 if not a stack)."""
return inspect_image(path).frames
def array_nbytes(path):
"""Bytes that opening this file's full array would allocate.
Computed from shape + dtype in the header - no pixel decode - so it is safe
to call on a file that is too large to open. Returns 0 if unknown.
"""
ext = os.path.splitext(path)[1].lower()
if ext in (".tif", ".tiff") and tifffile is not None:
try:
with tifffile.TiffFile(path) as tif:
series = tif.series[0]
return int(np.prod(series.shape)) * int(np.dtype(series.dtype).itemsize)
except Exception: # noqa: BLE001 - fall through to PIL
pass
try:
with Image.open(path) as img:
w, h = img.size
return int(w) * int(h) * len(img.getbands()) # 8-bit assumption
except Exception: # noqa: BLE001
return 0
def image_size(path):
"""Return an image's (width, height) by reading only its header.
Never decodes pixel data, so this is safe to call as a size guard on a file
that would be too large to load. Returns (0, 0) if the size cannot be
determined, so callers treat it as "unknown" and proceed.
"""
ext = os.path.splitext(path)[1].lower()
if ext in (".tif", ".tiff") and tifffile is not None:
try:
with tifffile.TiffFile(path) as tif:
page = tif.pages[0]
return int(page.imagewidth), int(page.imagelength)
except Exception: # noqa: BLE001 - fall through to PIL
pass
try:
with Image.open(path) as img: # PIL parses the header lazily
return int(img.size[0]), int(img.size[1])
except Exception: # noqa: BLE001
return 0, 0
def _to_rgb(arr):
"""Turn a 2D or (H, W, C) array into exactly 3 channels."""
if arr.ndim == 2:
return np.stack([arr] * 3, axis=-1)
channels = arr.shape[2]
if channels == 1:
return np.repeat(arr, 3, axis=2)
if channels == 2:
# Pad a zero third channel rather than inventing signal.
return np.concatenate([arr, np.zeros_like(arr[:, :, :1])], axis=2)
return arr[:, :, :3]
def _load_rgb(path, frame=0):
"""Read a file and reduce it to an (H, W, 3) array plus its source dtype."""
raw, axes = _read_array(path)
arr = _reduce_to_hwc(raw, axes, frame=frame)
arr = _to_rgb(arr)
return arr, raw.dtype
def _to_uint8(arr, stretch):
"""Map pixel values to uint8.
When ``stretch`` is True (non-8-bit input) a 1st-99th percentile auto-
contrast stretch is applied - outlier-robust, so a hot/saturated pixel does
not collapse the visible signal to black. Otherwise values are only clipped,
so standard 8-bit images are unchanged.
"""
arr = arr.astype(np.float32)
if not stretch:
return np.clip(arr, 0, 255).astype(np.uint8)
lo, hi = np.percentile(arr, (1, 99))
if hi <= lo: # near-flat image; fall back to full min/max
lo, hi = float(arr.min()), float(arr.max())
if hi <= lo: # truly constant image
return np.zeros(arr.shape, dtype=np.uint8)
arr = np.clip((arr - lo) / (hi - lo), 0.0, 1.0)
return (arr * 255.0).astype(np.uint8)
def _save_png(arr, out_path=None, out_dir=None, base=None, suffix="_std.png"):
"""Save an (H, W, 3) uint8 array as a PNG and return the path."""
if out_path is None:
if out_dir is not None:
os.makedirs(out_dir, exist_ok=True)
out_path = os.path.join(out_dir, (base or "image") + suffix)
else:
tmp = tempfile.NamedTemporaryFile(delete=False, suffix=suffix)
out_path = tmp.name
tmp.close()
Image.fromarray(arr, mode="RGB").save(out_path)
return out_path
def standardize_image(path, frame=0, out_dir=None, out_path=None):
"""Standardize any image/TIFF to a canonical 8-bit RGB PNG.
Used for both the preview and the model: one frame, <=3 channels, with a
1st-99th percentile auto-contrast stretch for 16-bit/float input (8-bit is
passed through unchanged).
Args:
path: path to the input image (TIFF, PNG, JPG, ...).
frame: which frame of a stack to use (0-based; ignored for non-stacks).
out_dir: optional directory for the output PNG.
out_path: optional explicit output file path (overrides out_dir).
Returns:
Path to the standardized PNG. On any failure the original ``path`` is
returned unchanged so callers degrade gracefully.
"""
try:
arr, dtype = _load_rgb(path, frame=frame)
arr = _to_uint8(arr, stretch=(dtype != np.uint8))
base = os.path.splitext(os.path.basename(path))[0]
return _save_png(arr, out_path=out_path, out_dir=out_dir, base=base, suffix="_std.png")
except Exception as e: # noqa: BLE001 - never let preprocessing crash a run
print(f"⚠️ standardize_image failed for {path}: {e}; using original file")
return path
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