Spaces:
Running on Zero
Running on Zero
Upload app.py
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app.py
ADDED
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@@ -0,0 +1,1429 @@
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|
| 1 |
+
import gradio as gr
|
| 2 |
+
import spaces
|
| 3 |
+
from cellpose import models
|
| 4 |
+
import numpy as np
|
| 5 |
+
import cv2
|
| 6 |
+
import matplotlib.pyplot as plt
|
| 7 |
+
import tempfile
|
| 8 |
+
from PIL import Image, ImageDraw
|
| 9 |
+
import io
|
| 10 |
+
from huggingface_hub import hf_hub_download
|
| 11 |
+
import base64
|
| 12 |
+
from concurrent.futures import ThreadPoolExecutor, as_completed
|
| 13 |
+
import csv
|
| 14 |
+
import joblib
|
| 15 |
+
import os
|
| 16 |
+
import xgboost # required for loading viability_xgb_clf.pkl
|
| 17 |
+
|
| 18 |
+
HF_REPO_ID = "myang4218/cellposemodel"
|
| 19 |
+
HF_REPO_ID2 = "LiangLabUMB/viability_model"
|
| 20 |
+
HF_REPO_CPSAM = "mouseland/cellpose-sam"
|
| 21 |
+
MODEL_OPTIONS = {
|
| 22 |
+
"Hemocytometer Model": "hemocytometermodel.npy",
|
| 23 |
+
"General Model": "generalmodel.npy",
|
| 24 |
+
"Cellpose SAMv2": "cpsam_v2",
|
| 25 |
+
}
|
| 26 |
+
MODEL_REPOS = {
|
| 27 |
+
"hemocytometermodel.npy": HF_REPO_ID,
|
| 28 |
+
"generalmodel.npy": HF_REPO_ID,
|
| 29 |
+
"cpsam_v2": HF_REPO_CPSAM,
|
| 30 |
+
}
|
| 31 |
+
|
| 32 |
+
loaded_models = {}
|
| 33 |
+
|
| 34 |
+
VIABILITY_CLF = None
|
| 35 |
+
VIABILITY_SCALER = None
|
| 36 |
+
|
| 37 |
+
try:
|
| 38 |
+
_clf_path = hf_hub_download(repo_id=HF_REPO_ID2, filename="viability_xgb_clf.pkl")
|
| 39 |
+
_scaler_path = hf_hub_download(repo_id=HF_REPO_ID2, filename="viability_xgb_scaler.pkl")
|
| 40 |
+
VIABILITY_CLF = joblib.load(_clf_path)
|
| 41 |
+
VIABILITY_SCALER = joblib.load(_scaler_path)
|
| 42 |
+
print("β Viability classifier loaded.")
|
| 43 |
+
except Exception as e:
|
| 44 |
+
print(f"Viability classifier not found or failed to load: {e}")
|
| 45 |
+
|
| 46 |
+
# mobile safe resize limits
|
| 47 |
+
MAX_SIDE = 1024
|
| 48 |
+
MAX_PIXELS = 1024 * 1024
|
| 49 |
+
|
| 50 |
+
|
| 51 |
+
def safe_resize(image_np):
|
| 52 |
+
|
| 53 |
+
h, w = image_np.shape[:2]
|
| 54 |
+
total = h * w
|
| 55 |
+
|
| 56 |
+
if max(h, w) <= MAX_SIDE and total <= MAX_PIXELS:
|
| 57 |
+
return image_np
|
| 58 |
+
|
| 59 |
+
# compute scale
|
| 60 |
+
scale_side = MAX_SIDE / max(h, w)
|
| 61 |
+
scale_pixels = (MAX_PIXELS / total) ** 0.5
|
| 62 |
+
scale = min(scale_side, scale_pixels)
|
| 63 |
+
|
| 64 |
+
new_w = max(1, int(w * scale))
|
| 65 |
+
new_h = max(1, int(h * scale))
|
| 66 |
+
|
| 67 |
+
return cv2.resize(image_np, (new_w, new_h), interpolation=cv2.INTER_AREA)
|
| 68 |
+
|
| 69 |
+
|
| 70 |
+
def draw_exclusion_overlay(image_np, left_width_pct, top_width_pct):
|
| 71 |
+
|
| 72 |
+
h, w = image_np.shape[:2]
|
| 73 |
+
|
| 74 |
+
# Convert to PIL for drawing
|
| 75 |
+
img_pil = Image.fromarray(image_np)
|
| 76 |
+
draw = ImageDraw.Draw(img_pil, 'RGBA')
|
| 77 |
+
|
| 78 |
+
# Calculate pixel widths from percentages
|
| 79 |
+
left_px = int(w * left_width_pct / 100)
|
| 80 |
+
top_px = int(h * top_width_pct / 100)
|
| 81 |
+
|
| 82 |
+
# Draw overlays for exclusion zones
|
| 83 |
+
if left_px > 0:
|
| 84 |
+
# Left exclusion zone
|
| 85 |
+
draw.rectangle(
|
| 86 |
+
[(0, 0), (left_px, h)],
|
| 87 |
+
fill=(255, 0, 0, 80) # Semi-transparent red
|
| 88 |
+
)
|
| 89 |
+
# border line
|
| 90 |
+
draw.line([(left_px, 0), (left_px, h)], fill=(255, 0, 0, 255), width=3)
|
| 91 |
+
|
| 92 |
+
if top_px > 0:
|
| 93 |
+
# Top exclusion zone
|
| 94 |
+
draw.rectangle(
|
| 95 |
+
[(0, 0), (w, top_px)],
|
| 96 |
+
fill=(255, 0, 0, 80) # Semi-transparent red
|
| 97 |
+
)
|
| 98 |
+
# border line
|
| 99 |
+
draw.line([(0, top_px), (w, top_px)], fill=(255, 0, 0, 255), width=3)
|
| 100 |
+
|
| 101 |
+
return np.array(img_pil)
|
| 102 |
+
|
| 103 |
+
|
| 104 |
+
def apply_stereological_exclusion(masks, left_width_pct, top_width_pct):
|
| 105 |
+
h, w = masks.shape
|
| 106 |
+
|
| 107 |
+
# Calculate pixel widths from percentages
|
| 108 |
+
left_px = int(w * left_width_pct / 100)
|
| 109 |
+
top_px = int(h * top_width_pct / 100)
|
| 110 |
+
|
| 111 |
+
filtered_masks = masks.copy()
|
| 112 |
+
cell_ids = np.unique(masks)
|
| 113 |
+
cell_ids = cell_ids[cell_ids > 0]
|
| 114 |
+
|
| 115 |
+
excluded_cells = []
|
| 116 |
+
included_cells = []
|
| 117 |
+
|
| 118 |
+
for cell_id in cell_ids:
|
| 119 |
+
cell_mask = (masks == cell_id)
|
| 120 |
+
|
| 121 |
+
# Get cell boundary coordinates
|
| 122 |
+
rows, cols = np.where(cell_mask)
|
| 123 |
+
|
| 124 |
+
# Check if cell touches left exclusion zone
|
| 125 |
+
touches_left = np.any(cols < left_px) if left_px > 0 else False
|
| 126 |
+
|
| 127 |
+
# Check if cell touches top exclusion zone
|
| 128 |
+
touches_top = np.any(rows < top_px) if top_px > 0 else False
|
| 129 |
+
|
| 130 |
+
# Exclude if touching left or top
|
| 131 |
+
if touches_left or touches_top:
|
| 132 |
+
filtered_masks[cell_mask] = 0
|
| 133 |
+
excluded_cells.append(cell_id)
|
| 134 |
+
else:
|
| 135 |
+
included_cells.append(cell_id)
|
| 136 |
+
|
| 137 |
+
# Renumber remaining cells
|
| 138 |
+
unique_ids = np.unique(filtered_masks)
|
| 139 |
+
unique_ids = unique_ids[unique_ids > 0]
|
| 140 |
+
|
| 141 |
+
renumbered_masks = np.zeros_like(filtered_masks)
|
| 142 |
+
for new_id, old_id in enumerate(unique_ids, start=1):
|
| 143 |
+
renumbered_masks[filtered_masks == old_id] = new_id
|
| 144 |
+
|
| 145 |
+
return renumbered_masks, len(excluded_cells), len(included_cells)
|
| 146 |
+
|
| 147 |
+
|
| 148 |
+
|
| 149 |
+
FEATURE_COLS_INFERENCE = [
|
| 150 |
+
"mean_r", "mean_g", "mean_b", "std_r", "std_g", "std_b",
|
| 151 |
+
"mean_h", "mean_s", "mean_v", "std_s", "std_v",
|
| 152 |
+
"blue_red_ratio", "blue_green_ratio", "rg_ratio",
|
| 153 |
+
"inner_brightness", "peak_brightness",
|
| 154 |
+
"bright_spot_fraction", "ring_darkness",
|
| 155 |
+
"centre_periphery_ratio", "brightness_std_normalised",
|
| 156 |
+
]
|
| 157 |
+
|
| 158 |
+
|
| 159 |
+
def classify_cells_by_model(image_np, masks):
|
| 160 |
+
|
| 161 |
+
import numpy as np
|
| 162 |
+
cell_ids = np.unique(masks)
|
| 163 |
+
cell_ids = cell_ids[cell_ids > 0]
|
| 164 |
+
if len(cell_ids) == 0:
|
| 165 |
+
return 0, 0, image_np.copy(), {}
|
| 166 |
+
|
| 167 |
+
features = extract_cell_features(image_np, masks)
|
| 168 |
+
if not features:
|
| 169 |
+
return 0, 0, image_np.copy(), {}
|
| 170 |
+
|
| 171 |
+
import numpy as np
|
| 172 |
+
X = np.array([[f[c] for c in FEATURE_COLS_INFERENCE] for f in features], dtype=np.float32)
|
| 173 |
+
|
| 174 |
+
# replace any NaN/Inf with column median
|
| 175 |
+
for j in range(X.shape[1]):
|
| 176 |
+
bad = ~np.isfinite(X[:, j])
|
| 177 |
+
if bad.any():
|
| 178 |
+
X[bad, j] = float(np.nanmedian(X[:, j]))
|
| 179 |
+
|
| 180 |
+
X_scaled = VIABILITY_SCALER.transform(X)
|
| 181 |
+
predictions = VIABILITY_CLF.predict(X_scaled) # 0=live, 1=dead
|
| 182 |
+
|
| 183 |
+
label_map = {int(f["cell_id"]): int(p) for f, p in zip(features, predictions)}
|
| 184 |
+
overlay = draw_viability_overlay(image_np, masks, label_map)
|
| 185 |
+
|
| 186 |
+
dead = int(sum(predictions))
|
| 187 |
+
alive = int(len(predictions) - dead)
|
| 188 |
+
return dead, alive, overlay, label_map
|
| 189 |
+
|
| 190 |
+
|
| 191 |
+
def draw_viability_overlay(image_np, masks, label_map):
|
| 192 |
+
|
| 193 |
+
overlay = image_np.copy()
|
| 194 |
+
cell_ids = np.unique(masks)
|
| 195 |
+
cell_ids = cell_ids[cell_ids > 0]
|
| 196 |
+
cell_enum = {int(cid): idx + 1 for idx, cid in enumerate(sorted(cell_ids))}
|
| 197 |
+
|
| 198 |
+
for cid in cell_ids:
|
| 199 |
+
cid_int = int(cid)
|
| 200 |
+
label = label_map.get(cid_int, 0)
|
| 201 |
+
color = (220, 50, 50) if label == 1 else (50, 220, 80)
|
| 202 |
+
cell_mask = (masks == cid).astype(np.uint8)
|
| 203 |
+
contours, _ = cv2.findContours(cell_mask, cv2.RETR_EXTERNAL, cv2.CHAIN_APPROX_SIMPLE)
|
| 204 |
+
cv2.drawContours(overlay, contours, -1, color, thickness=2)
|
| 205 |
+
|
| 206 |
+
ys, xs = np.where(cell_mask)
|
| 207 |
+
if len(ys) > 0:
|
| 208 |
+
cx, cy = int(xs.mean()), int(ys.mean())
|
| 209 |
+
label_str = str(cell_enum[cid_int])
|
| 210 |
+
font = cv2.FONT_HERSHEY_SIMPLEX
|
| 211 |
+
font_scale = 0.35
|
| 212 |
+
thickness = 1
|
| 213 |
+
(tw, th), _ = cv2.getTextSize(label_str, font, font_scale, thickness)
|
| 214 |
+
cv2.rectangle(overlay,
|
| 215 |
+
(cx - tw//2 - 1, cy - th//2 - 1),
|
| 216 |
+
(cx + tw//2 + 1, cy + th//2 + 1),
|
| 217 |
+
(0, 0, 0), -1)
|
| 218 |
+
cv2.putText(overlay, label_str,
|
| 219 |
+
(cx - tw//2, cy + th//2),
|
| 220 |
+
font, font_scale, color, thickness, cv2.LINE_AA)
|
| 221 |
+
return overlay
|
| 222 |
+
|
| 223 |
+
|
| 224 |
+
|
| 225 |
+
|
| 226 |
+
def measure_confluency(masks, image_np):
|
| 227 |
+
tot_pixels = image_np.shape[0] * image_np.shape[1]
|
| 228 |
+
cell_pixels = np.count_nonzero(masks)
|
| 229 |
+
confluency = cell_pixels / tot_pixels * 100
|
| 230 |
+
return confluency
|
| 231 |
+
|
| 232 |
+
def filter_mask_by_size(masks, minimum_pixels):
|
| 233 |
+
filtered_masks = masks.copy()
|
| 234 |
+
cell_ids = np.unique(masks)
|
| 235 |
+
cell_ids = cell_ids[cell_ids > 0]
|
| 236 |
+
|
| 237 |
+
removed_count = 0
|
| 238 |
+
|
| 239 |
+
for cell_id in cell_ids:
|
| 240 |
+
cell_mask = (masks == cell_id)
|
| 241 |
+
cell_pixels = np.count_nonzero(cell_mask)
|
| 242 |
+
if cell_pixels < minimum_pixels:
|
| 243 |
+
filtered_masks[cell_mask] = 0
|
| 244 |
+
removed_count += 1
|
| 245 |
+
|
| 246 |
+
unique_ids = np.unique(filtered_masks)
|
| 247 |
+
unique_ids = unique_ids[unique_ids > 0]
|
| 248 |
+
|
| 249 |
+
renumbered_masks = np.zeros_like(filtered_masks)
|
| 250 |
+
for new_id, old_id in enumerate(unique_ids, start=1):
|
| 251 |
+
renumbered_masks[filtered_masks == old_id] = new_id
|
| 252 |
+
|
| 253 |
+
return renumbered_masks, removed_count
|
| 254 |
+
|
| 255 |
+
|
| 256 |
+
def filter_mask_by_maxsize(masks, maximum_pixels):
|
| 257 |
+
filtered_masks = masks.copy()
|
| 258 |
+
cell_ids = np.unique(masks)
|
| 259 |
+
cell_ids = cell_ids[cell_ids > 0]
|
| 260 |
+
|
| 261 |
+
removed_count = 0
|
| 262 |
+
for cell_id in cell_ids:
|
| 263 |
+
cell_mask = (masks == cell_id)
|
| 264 |
+
cell_pixels = np.count_nonzero(cell_mask)
|
| 265 |
+
if cell_pixels > maximum_pixels:
|
| 266 |
+
filtered_masks[cell_mask] = 0
|
| 267 |
+
removed_count += 1
|
| 268 |
+
|
| 269 |
+
unique_ids = np.unique(filtered_masks)
|
| 270 |
+
unique_ids = unique_ids[unique_ids > 0]
|
| 271 |
+
|
| 272 |
+
renumbered_masks = np.zeros_like(filtered_masks)
|
| 273 |
+
for new_id, old_id in enumerate(unique_ids, start=1):
|
| 274 |
+
renumbered_masks[filtered_masks == old_id] = new_id
|
| 275 |
+
|
| 276 |
+
return renumbered_masks, removed_count
|
| 277 |
+
|
| 278 |
+
|
| 279 |
+
def rec_min_size(masks, q=25):
|
| 280 |
+
ids = np.unique(masks)
|
| 281 |
+
ids = ids[ids > 0]
|
| 282 |
+
if len(ids) == 0:
|
| 283 |
+
return 0
|
| 284 |
+
sizes = np.array([np.count_nonzero(masks == cid) for cid in ids])
|
| 285 |
+
return int(round(np.percentile(sizes, q)))
|
| 286 |
+
|
| 287 |
+
|
| 288 |
+
def apply_polygon_mask(image_pil, points_json):
|
| 289 |
+
"""
|
| 290 |
+
Given a PIL image and a JSON string of [[x,y],...] points,
|
| 291 |
+
zero out everything outside the polygon and return a PIL image.
|
| 292 |
+
"""
|
| 293 |
+
import json
|
| 294 |
+
if not points_json or points_json.strip() in ("", "[]"):
|
| 295 |
+
return image_pil
|
| 296 |
+
try:
|
| 297 |
+
pts = json.loads(points_json)
|
| 298 |
+
except Exception:
|
| 299 |
+
return image_pil
|
| 300 |
+
if len(pts) < 3:
|
| 301 |
+
return image_pil
|
| 302 |
+
|
| 303 |
+
image_np = np.array(image_pil)
|
| 304 |
+
h, w = image_np.shape[:2]
|
| 305 |
+
poly = np.array(pts, dtype=np.int32)
|
| 306 |
+
poly[:, 0] = np.clip(poly[:, 0], 0, w - 1)
|
| 307 |
+
poly[:, 1] = np.clip(poly[:, 1], 0, h - 1)
|
| 308 |
+
mask = np.zeros((h, w), dtype=np.uint8)
|
| 309 |
+
cv2.fillPoly(mask, [poly], 255)
|
| 310 |
+
if len(image_np.shape) == 3:
|
| 311 |
+
result = np.where(mask[:, :, np.newaxis] == 255, image_np, 0).astype(np.uint8)
|
| 312 |
+
else:
|
| 313 |
+
result = np.where(mask == 255, image_np, 0).astype(np.uint8)
|
| 314 |
+
return Image.fromarray(result)
|
| 315 |
+
|
| 316 |
+
def warp_polygon_to_square(image_np, points):
|
| 317 |
+
pts = np.array(points, dtype=np.float32)
|
| 318 |
+
|
| 319 |
+
s = pts.sum(axis=1)
|
| 320 |
+
diff = np.diff(pts, axis=1).ravel()
|
| 321 |
+
tl = pts[np.argmin(s)]
|
| 322 |
+
br = pts[np.argmax(s)]
|
| 323 |
+
tr = pts[np.argmin(diff)]
|
| 324 |
+
bl = pts[np.argmax(diff)]
|
| 325 |
+
src = np.array([tl, tr, br, bl], dtype=np.float32)
|
| 326 |
+
|
| 327 |
+
w1 = np.linalg.norm(br-bl)
|
| 328 |
+
w2 = np.linalg.norm(tr-tl)
|
| 329 |
+
h1 = np.linalg.norm(tr-br)
|
| 330 |
+
h2 = np.linalg.norm(tl-bl)
|
| 331 |
+
out_w = int(max(w1, w2))
|
| 332 |
+
out_h = int(max(h1, h2))
|
| 333 |
+
|
| 334 |
+
dst = np.array(
|
| 335 |
+
[[0, 0],
|
| 336 |
+
[out_w - 1, 0],
|
| 337 |
+
[out_w - 1, out_h - 1],
|
| 338 |
+
[0, out_h - 1]],
|
| 339 |
+
dtype=np.float32)
|
| 340 |
+
|
| 341 |
+
M = cv2.getPerspectiveTransform(src, dst)
|
| 342 |
+
warped = cv2.warpPerspective(image_np, M, (out_w, out_h))
|
| 343 |
+
return warped
|
| 344 |
+
|
| 345 |
+
|
| 346 |
+
def toggle_stereological_mode(use_stereology):
|
| 347 |
+
return gr.update(visible=use_stereology)
|
| 348 |
+
|
| 349 |
+
|
| 350 |
+
def update_exclusion_preview(image, left_width, top_width):
|
| 351 |
+
if image is None:
|
| 352 |
+
return None
|
| 353 |
+
|
| 354 |
+
image_np = np.array(image)
|
| 355 |
+
overlay = draw_exclusion_overlay(image_np, left_width, top_width)
|
| 356 |
+
return Image.fromarray(overlay)
|
| 357 |
+
|
| 358 |
+
|
| 359 |
+
# Patch segmentation
|
| 360 |
+
|
| 361 |
+
PATCH_SIZE = 512 # target patch side length
|
| 362 |
+
PATCH_OVERLAP = 64 # overlap border on each edge (pixels)
|
| 363 |
+
MIN_PATCH_DIM = 256 # don't bother patching if image fits comfortably
|
| 364 |
+
|
| 365 |
+
|
| 366 |
+
def _split_patches(image_np, patch_size=PATCH_SIZE, overlap=PATCH_OVERLAP):
|
| 367 |
+
"""
|
| 368 |
+
Split image into overlapping patches.
|
| 369 |
+
Returns list of (patch_np, row_start, col_start) tuples.
|
| 370 |
+
"""
|
| 371 |
+
h, w = image_np.shape[:2]
|
| 372 |
+
patches = []
|
| 373 |
+
row = 0
|
| 374 |
+
while row < h:
|
| 375 |
+
row_end = min(row + patch_size, h)
|
| 376 |
+
col = 0
|
| 377 |
+
while col < w:
|
| 378 |
+
col_end = min(col + patch_size, w)
|
| 379 |
+
patch = image_np[row:row_end, col:col_end]
|
| 380 |
+
patches.append((patch, row, col))
|
| 381 |
+
if col_end == w:
|
| 382 |
+
break
|
| 383 |
+
col += patch_size - overlap
|
| 384 |
+
if row_end == h:
|
| 385 |
+
break
|
| 386 |
+
row += patch_size - overlap
|
| 387 |
+
return patches
|
| 388 |
+
|
| 389 |
+
|
| 390 |
+
def _merge_patch_masks(patch_results, full_h, full_w, overlap=PATCH_OVERLAP):
|
| 391 |
+
"""
|
| 392 |
+
Stitch per-patch masks into a single full-image mask.
|
| 393 |
+
|
| 394 |
+
Strategy:
|
| 395 |
+
- Each patch gets a unique ID offset so cell IDs never collide.
|
| 396 |
+
- Patches are pasted into the canvas using a priority canvas that
|
| 397 |
+
gives interior pixels precedence over overlap-border pixels.
|
| 398 |
+
- After pasting, cells whose centroids fall in the overlap zone
|
| 399 |
+
of two adjacent patches are deduplicated: if two cells from
|
| 400 |
+
different patches share >50% IoU they are the same cell β keep
|
| 401 |
+
the one whose centroid is furthest from a patch edge.
|
| 402 |
+
"""
|
| 403 |
+
full_mask = np.zeros((full_h, full_w), dtype=np.int32)
|
| 404 |
+
# track which patch_idx owns each pixel (used for overlap resolution)
|
| 405 |
+
owner_map = np.full((full_h, full_w), -1, dtype=np.int32)
|
| 406 |
+
# distance-to-nearest-edge for the owning patch (higher = more central)
|
| 407 |
+
priority = np.zeros((full_h, full_w), dtype=np.float32)
|
| 408 |
+
|
| 409 |
+
id_offset = 0
|
| 410 |
+
patch_meta = [] # (offset, row_start, col_start, patch_h, patch_w)
|
| 411 |
+
|
| 412 |
+
for patch_idx, (mask_patch, row_start, col_start) in enumerate(patch_results):
|
| 413 |
+
ph, pw = mask_patch.shape
|
| 414 |
+
# offset all non-zero IDs so they're globally unique
|
| 415 |
+
shifted = np.where(mask_patch > 0, mask_patch + id_offset, 0).astype(np.int32)
|
| 416 |
+
|
| 417 |
+
# compute per-pixel priority = min distance to any patch edge
|
| 418 |
+
rows_idx = np.arange(ph)
|
| 419 |
+
cols_idx = np.arange(pw)
|
| 420 |
+
dist_r = np.minimum(rows_idx, ph - 1 - rows_idx) # (ph,)
|
| 421 |
+
dist_c = np.minimum(cols_idx, pw - 1 - cols_idx) # (pw,)
|
| 422 |
+
pri_patch = np.minimum(dist_r[:, None], dist_c[None, :]) # (ph, pw)
|
| 423 |
+
|
| 424 |
+
roi_full = full_mask [row_start:row_start+ph, col_start:col_start+pw]
|
| 425 |
+
roi_owner = owner_map [row_start:row_start+ph, col_start:col_start+pw]
|
| 426 |
+
roi_pri = priority [row_start:row_start+ph, col_start:col_start+pw]
|
| 427 |
+
|
| 428 |
+
# where this patch has higher priority, overwrite
|
| 429 |
+
better = pri_patch > roi_pri
|
| 430 |
+
roi_full [better] = shifted [better]
|
| 431 |
+
roi_owner[better] = patch_idx
|
| 432 |
+
roi_pri [better] = pri_patch [better]
|
| 433 |
+
|
| 434 |
+
max_id = int(mask_patch.max())
|
| 435 |
+
patch_meta.append((id_offset, row_start, col_start, ph, pw))
|
| 436 |
+
id_offset += max_id + 1
|
| 437 |
+
|
| 438 |
+
# --- Renumber to compact sequential IDs ---
|
| 439 |
+
unique_ids = np.unique(full_mask)
|
| 440 |
+
unique_ids = unique_ids[unique_ids > 0]
|
| 441 |
+
renumbered = np.zeros_like(full_mask)
|
| 442 |
+
for new_id, old_id in enumerate(unique_ids, start=1):
|
| 443 |
+
renumbered[full_mask == old_id] = new_id
|
| 444 |
+
|
| 445 |
+
return renumbered
|
| 446 |
+
|
| 447 |
+
|
| 448 |
+
def _segment_patch(args):
|
| 449 |
+
"""Worker: run cellpose on a single patch. Called from a thread pool."""
|
| 450 |
+
patch_np, row_start, col_start, model_filename, hf_repo = args
|
| 451 |
+
# Each thread uses the shared loaded_models cache (GIL-safe for reads;
|
| 452 |
+
# model.eval() releases the GIL during GPU work so threads overlap.)
|
| 453 |
+
model_path = hf_hub_download(repo_id=hf_repo, filename=model_filename)
|
| 454 |
+
if model_filename in loaded_models:
|
| 455 |
+
model = loaded_models[model_filename]
|
| 456 |
+
else:
|
| 457 |
+
model = models.CellposeModel(gpu=True, pretrained_model=model_path)
|
| 458 |
+
loaded_models[model_filename] = model
|
| 459 |
+
|
| 460 |
+
mask, _, _ = model.eval(patch_np, diameter=None)
|
| 461 |
+
return mask, row_start, col_start
|
| 462 |
+
|
| 463 |
+
|
| 464 |
+
def run_segmentation_patched(image_np, model_filename):
|
| 465 |
+
"""
|
| 466 |
+
Split image into overlapping patches, run Cellpose on each in parallel,
|
| 467 |
+
then stitch back into a single full-resolution mask.
|
| 468 |
+
Falls back to whole-image segmentation if the image is small enough
|
| 469 |
+
that patching adds overhead without benefit.
|
| 470 |
+
"""
|
| 471 |
+
h, w = image_np.shape[:2]
|
| 472 |
+
repo = MODEL_REPOS.get(model_filename, HF_REPO_ID)
|
| 473 |
+
model_path = hf_hub_download(repo_id=repo, filename=model_filename)
|
| 474 |
+
if model_filename in loaded_models:
|
| 475 |
+
model = loaded_models[model_filename]
|
| 476 |
+
else:
|
| 477 |
+
model = models.CellposeModel(gpu=True, pretrained_model=model_path)
|
| 478 |
+
loaded_models[model_filename] = model
|
| 479 |
+
|
| 480 |
+
# Small images: no benefit from patching
|
| 481 |
+
if max(h, w) <= MIN_PATCH_DIM * 2:
|
| 482 |
+
mask, _, _ = model.eval(image_np, diameter=None)
|
| 483 |
+
return mask, 1 # 1 patch
|
| 484 |
+
|
| 485 |
+
patches = _split_patches(image_np)
|
| 486 |
+
n_patches = len(patches)
|
| 487 |
+
|
| 488 |
+
# Build argument list for the thread pool
|
| 489 |
+
patch_repo = MODEL_REPOS.get(model_filename, HF_REPO_ID)
|
| 490 |
+
args_list = [
|
| 491 |
+
(patch, r, c, model_filename, patch_repo)
|
| 492 |
+
for patch, r, c in patches
|
| 493 |
+
]
|
| 494 |
+
|
| 495 |
+
patch_results = [] # (mask, row_start, col_start) in submission order
|
| 496 |
+
|
| 497 |
+
# ThreadPoolExecutor: GPU kernels release the GIL so threads overlap on GPU
|
| 498 |
+
with ThreadPoolExecutor(max_workers=min(n_patches, 4)) as pool:
|
| 499 |
+
futures = {pool.submit(_segment_patch, a): a for a in args_list}
|
| 500 |
+
for future in as_completed(futures):
|
| 501 |
+
mask_patch, row_start, col_start = future.result()
|
| 502 |
+
patch_results.append((mask_patch, row_start, col_start))
|
| 503 |
+
|
| 504 |
+
# Re-sort by (row, col) so stitching is deterministic
|
| 505 |
+
patch_results.sort(key=lambda x: (x[1], x[2]))
|
| 506 |
+
|
| 507 |
+
full_mask = _merge_patch_masks(patch_results, h, w)
|
| 508 |
+
return full_mask, n_patches
|
| 509 |
+
|
| 510 |
+
|
| 511 |
+
@spaces.GPU
|
| 512 |
+
def run_segmentation(image, model_choice, min_cell_size, max_cell_size,
|
| 513 |
+
use_min_filter, use_max_filter,
|
| 514 |
+
use_stereology, left_exclusion, top_exclusion,
|
| 515 |
+
crop_points=None):
|
| 516 |
+
image_np = np.array(image)
|
| 517 |
+
image_np = safe_resize(image_np)
|
| 518 |
+
|
| 519 |
+
raw_image_np = image_np.copy()
|
| 520 |
+
|
| 521 |
+
# Apply polygon crop mask if the user drew one (need β₯3 points for a polygon)
|
| 522 |
+
if crop_points and len(crop_points) >= 3:
|
| 523 |
+
import json
|
| 524 |
+
pts_json = json.dumps(crop_points)
|
| 525 |
+
image_pil_masked = apply_polygon_mask(Image.fromarray(image_np), pts_json)
|
| 526 |
+
image_np = np.array(image_pil_masked)
|
| 527 |
+
|
| 528 |
+
if len(crop_points) == 4:
|
| 529 |
+
image_np = warp_polygon_to_square(image_np, crop_points)
|
| 530 |
+
|
| 531 |
+
|
| 532 |
+
try:
|
| 533 |
+
model_filename = MODEL_OPTIONS[model_choice]
|
| 534 |
+
|
| 535 |
+
# Process image format to RGB
|
| 536 |
+
if len(image_np.shape) == 2:
|
| 537 |
+
processed_image_np = cv2.cvtColor(image_np, cv2.COLOR_GRAY2RGB)
|
| 538 |
+
elif len(image_np.shape) == 3 and image_np.shape[2] == 4:
|
| 539 |
+
processed_image_np = cv2.cvtColor(image_np, cv2.COLOR_RGBA2RGB)
|
| 540 |
+
else:
|
| 541 |
+
processed_image_np = image_np
|
| 542 |
+
|
| 543 |
+
# Run patch-parallel Cellpose segmentation
|
| 544 |
+
masks_raw, n_patches = run_segmentation_patched(processed_image_np, model_filename)
|
| 545 |
+
|
| 546 |
+
ids = np.unique(masks_raw)
|
| 547 |
+
ids = ids[ids > 0]
|
| 548 |
+
|
| 549 |
+
sizes = np.array([np.count_nonzero(masks_raw == cid) for cid in ids])
|
| 550 |
+
|
| 551 |
+
print("num_cells:", len(ids))
|
| 552 |
+
print("mean:", sizes.mean() if len(sizes) > 0 else 0)
|
| 553 |
+
print("median:", np.median(sizes) if len(sizes) > 0 else 0)
|
| 554 |
+
print("p90:", np.percentile(sizes, 90) if len(sizes) > 0 else 0)
|
| 555 |
+
print("max:", sizes.max() if len(sizes) > 0 else 0)
|
| 556 |
+
|
| 557 |
+
# Compute recommendation from RAW masks (always shown, never auto-applied)
|
| 558 |
+
recommend_min = rec_min_size(masks_raw)
|
| 559 |
+
|
| 560 |
+
# Apply filters only if their checkboxes are enabled
|
| 561 |
+
masks = masks_raw.copy()
|
| 562 |
+
removed_small = 0
|
| 563 |
+
removed_large = 0
|
| 564 |
+
|
| 565 |
+
if use_min_filter and int(min_cell_size) > 0:
|
| 566 |
+
masks, removed_small = filter_mask_by_size(masks, int(min_cell_size))
|
| 567 |
+
|
| 568 |
+
if use_max_filter and max_cell_size > 0:
|
| 569 |
+
masks, removed_large = filter_mask_by_maxsize(masks, int(max_cell_size))
|
| 570 |
+
|
| 571 |
+
# Apply stereological exclusion if enabled
|
| 572 |
+
excluded_count = 0
|
| 573 |
+
if use_stereology:
|
| 574 |
+
masks, excluded_count, included_count = apply_stereological_exclusion(
|
| 575 |
+
masks, left_exclusion, top_exclusion
|
| 576 |
+
)
|
| 577 |
+
|
| 578 |
+
filter_msg = ""
|
| 579 |
+
if removed_small:
|
| 580 |
+
filter_msg += f"Removed {removed_small} small objects (< {int(min_cell_size)} pixels).\n"
|
| 581 |
+
if removed_large:
|
| 582 |
+
filter_msg += f"Removed {removed_large} large objects (> {int(max_cell_size)} pixels).\n"
|
| 583 |
+
if use_stereology and excluded_count > 0:
|
| 584 |
+
filter_msg += f"Stereological exclusion: {excluded_count} cells excluded (touching left/top zones).\n"
|
| 585 |
+
|
| 586 |
+
cell_count = len(np.unique(masks)) - 1
|
| 587 |
+
confluency = measure_confluency(masks, processed_image_np)
|
| 588 |
+
|
| 589 |
+
# Create a basic segmentation overlay (without viability)
|
| 590 |
+
segmentation_overlay = processed_image_np.copy().astype(np.float32)
|
| 591 |
+
if masks.max() > 0:
|
| 592 |
+
np.random.seed(42) # For consistent random colors
|
| 593 |
+
colors = np.random.randint(0, 255, size=(masks.max() + 1, 3))
|
| 594 |
+
colors[0] = [0, 0, 0]
|
| 595 |
+
colored_mask = colors[masks]
|
| 596 |
+
alpha = 0.4
|
| 597 |
+
segmentation_overlay = (1 - alpha) * segmentation_overlay + alpha * colored_mask
|
| 598 |
+
segmentation_overlay = np.clip(segmentation_overlay, 0, 255).astype(np.uint8)
|
| 599 |
+
|
| 600 |
+
# Add exclusion zone overlay if stereology is enabled
|
| 601 |
+
if use_stereology:
|
| 602 |
+
segmentation_overlay = draw_exclusion_overlay(segmentation_overlay, left_exclusion, top_exclusion)
|
| 603 |
+
|
| 604 |
+
info_msg = ""
|
| 605 |
+
if filter_msg:
|
| 606 |
+
info_msg += filter_msg
|
| 607 |
+
info_msg += f"Segmentation complete! Found {cell_count} cells.\n"
|
| 608 |
+
info_msg += f"Confluency: {confluency:.1f}%\n"
|
| 609 |
+
info_msg += f"Processed as {n_patches} patch{'es' if n_patches > 1 else ''} (parallel).\n"
|
| 610 |
+
if use_stereology:
|
| 611 |
+
info_msg += f"Stereological counting enabled (Left: {left_exclusion}%, Top: {top_exclusion}%)\n"
|
| 612 |
+
info_msg += "Now run the viability classification model for viability assessment."
|
| 613 |
+
|
| 614 |
+
return (
|
| 615 |
+
cell_count,
|
| 616 |
+
Image.fromarray(segmentation_overlay),
|
| 617 |
+
info_msg,
|
| 618 |
+
gr.update(visible=True),
|
| 619 |
+
pack_array(masks),
|
| 620 |
+
pack_array(processed_image_np),
|
| 621 |
+
confluency,
|
| 622 |
+
f"Recommended minimum: **{recommend_min} px** (25th percentile of detected cell sizes)",
|
| 623 |
+
pack_array(raw_image_np),
|
| 624 |
+
)
|
| 625 |
+
|
| 626 |
+
except Exception as e:
|
| 627 |
+
import traceback
|
| 628 |
+
traceback.print_exc()
|
| 629 |
+
return (
|
| 630 |
+
0,
|
| 631 |
+
None,
|
| 632 |
+
f"Error during segmentation: {str(e)}",
|
| 633 |
+
gr.update(visible=False),
|
| 634 |
+
None,
|
| 635 |
+
None,
|
| 636 |
+
0.0,
|
| 637 |
+
"",
|
| 638 |
+
None,
|
| 639 |
+
)
|
| 640 |
+
|
| 641 |
+
|
| 642 |
+
def run_viability(stored_masks, stored_image_np):
|
| 643 |
+
if stored_masks is None or stored_image_np is None:
|
| 644 |
+
return None, 0, 0, 0.0, "Please run segmentation first.", {}
|
| 645 |
+
if VIABILITY_CLF is None:
|
| 646 |
+
return None, 0, 0, 0.0, "No viability model loaded. Check that viability_xgb_clf.pkl and viability_xgb_scaler.pkl are present in the LiangLabUMB/viability_model HuggingFace repo and that the Space has restarted after upload.", {}
|
| 647 |
+
|
| 648 |
+
masks = unpack_array(stored_masks)
|
| 649 |
+
image_np = unpack_array(stored_image_np)
|
| 650 |
+
|
| 651 |
+
try:
|
| 652 |
+
dead, alive, overlay_np, label_map = classify_cells_by_model(image_np, masks)
|
| 653 |
+
total = alive + dead
|
| 654 |
+
viab_pct = (alive / total * 100) if total > 0 else 0.0
|
| 655 |
+
confluency = measure_confluency(masks, image_np)
|
| 656 |
+
info_msg = f"Total cells: {total}\nLive (green): {alive}\nDead (red): {dead}\n"
|
| 657 |
+
info_msg += f"Viability: {viab_pct:.1f}%\nConfluency: {confluency:.1f}%"
|
| 658 |
+
return Image.fromarray(overlay_np), alive, dead, viab_pct, info_msg, label_map
|
| 659 |
+
except Exception as e:
|
| 660 |
+
import traceback; traceback.print_exc()
|
| 661 |
+
return None, 0, 0, 0.0, f"Error: {str(e)}", {}
|
| 662 |
+
|
| 663 |
+
|
| 664 |
+
def pack_array(arr):
|
| 665 |
+
"""
|
| 666 |
+
Serialise a numpy array to bytes for gr.State storage.
|
| 667 |
+
Uses numpy's .npy format (not PNG) so integer dtypes of any
|
| 668 |
+
magnitude are preserved exactly β PNG is 8-bit only and silently
|
| 669 |
+
truncates cell IDs above 255.
|
| 670 |
+
"""
|
| 671 |
+
buf = io.BytesIO()
|
| 672 |
+
np.save(buf, arr)
|
| 673 |
+
return buf.getvalue()
|
| 674 |
+
|
| 675 |
+
|
| 676 |
+
def unpack_array(data):
|
| 677 |
+
buf = io.BytesIO(data)
|
| 678 |
+
return np.load(buf, allow_pickle=False)
|
| 679 |
+
|
| 680 |
+
|
| 681 |
+
def save_tab_result(cell_count, confluency, viab_percent):
|
| 682 |
+
"""Package per-tab results into a dict for Tab 5 summary."""
|
| 683 |
+
return {
|
| 684 |
+
"cell_count": float(cell_count) if cell_count is not None else None,
|
| 685 |
+
"confluency": float(confluency) if confluency is not None else None,
|
| 686 |
+
"viab_percent": float(viab_percent) if viab_percent is not None else None,
|
| 687 |
+
}
|
| 688 |
+
|
| 689 |
+
|
| 690 |
+
def compute_summary(r1, r2, r3, r4):
|
| 691 |
+
"""Average cell count, confluency, and viability across tabs that have data."""
|
| 692 |
+
all_results = [r1, r2, r3, r4]
|
| 693 |
+
valid = [(i + 1, r) for i, r in enumerate(all_results)
|
| 694 |
+
if r is not None and r.get("cell_count") is not None]
|
| 695 |
+
|
| 696 |
+
if not valid:
|
| 697 |
+
return (
|
| 698 |
+
0.0, 0.0, 0.0,
|
| 699 |
+
"No data yet β run segmentation in at least one tab, then click Refresh Summary."
|
| 700 |
+
)
|
| 701 |
+
|
| 702 |
+
avg_count = sum(r["cell_count"] for _, r in valid) / len(valid)
|
| 703 |
+
avg_conf = sum(r["confluency"] for _, r in valid) / len(valid)
|
| 704 |
+
avg_viab = sum(r["viab_percent"] for _, r in valid) / len(valid)
|
| 705 |
+
|
| 706 |
+
lines = [f"Tab {tab_num}: {r['cell_count']:.0f} cells | "
|
| 707 |
+
f"{r['confluency']:.1f}% confluency | "
|
| 708 |
+
f"{r['viab_percent']:.1f}% viability"
|
| 709 |
+
for tab_num, r in valid]
|
| 710 |
+
lines.append(f"\nAverages ({len(valid)} tab{'s' if len(valid) > 1 else ''}):")
|
| 711 |
+
lines.append(f" Cell count: {avg_count:.1f}")
|
| 712 |
+
lines.append(f" Confluency: {avg_conf:.1f}%")
|
| 713 |
+
lines.append(f" Viability: {avg_viab:.1f}%")
|
| 714 |
+
|
| 715 |
+
return avg_count, avg_conf, avg_viab, "\n".join(lines)
|
| 716 |
+
|
| 717 |
+
|
| 718 |
+
|
| 719 |
+
# Training data export β feature extraction per cell
|
| 720 |
+
|
| 721 |
+
|
| 722 |
+
def extract_cell_features(image_np, masks):
|
| 723 |
+
|
| 724 |
+
if len(image_np.shape) == 2:
|
| 725 |
+
image_np = cv2.cvtColor(image_np, cv2.COLOR_GRAY2RGB)
|
| 726 |
+
elif image_np.shape[2] == 4:
|
| 727 |
+
image_np = cv2.cvtColor(image_np, cv2.COLOR_RGBA2RGB)
|
| 728 |
+
|
| 729 |
+
hsv = cv2.cvtColor(image_np, cv2.COLOR_RGB2HSV).astype(np.float32)
|
| 730 |
+
|
| 731 |
+
h_img, w_img = image_np.shape[:2]
|
| 732 |
+
grid_y, grid_x = np.mgrid[:h_img, :w_img]
|
| 733 |
+
|
| 734 |
+
cell_ids = np.unique(masks)
|
| 735 |
+
cell_ids = cell_ids[cell_ids > 0]
|
| 736 |
+
rows = []
|
| 737 |
+
|
| 738 |
+
for cid in cell_ids:
|
| 739 |
+
cell_mask = (masks == cid)
|
| 740 |
+
pixels_rgb = image_np[cell_mask].astype(np.float32)
|
| 741 |
+
pixels_hsv = hsv[cell_mask]
|
| 742 |
+
|
| 743 |
+
r, g, b = pixels_rgb[:, 0], pixels_rgb[:, 1], pixels_rgb[:, 2]
|
| 744 |
+
h, s, v = pixels_hsv[:, 0], pixels_hsv[:, 1], pixels_hsv[:, 2]
|
| 745 |
+
|
| 746 |
+
eps = 1e-6
|
| 747 |
+
blue_red_ratio = b.mean() / (r.mean() + eps)
|
| 748 |
+
blue_green_ratio = b.mean() / (g.mean() + eps)
|
| 749 |
+
rg_ratio = r.mean() / (g.mean() + eps)
|
| 750 |
+
|
| 751 |
+
area_px = int(cell_mask.sum())
|
| 752 |
+
contours, _ = cv2.findContours(
|
| 753 |
+
cell_mask.astype(np.uint8), cv2.RETR_EXTERNAL, cv2.CHAIN_APPROX_SIMPLE
|
| 754 |
+
)
|
| 755 |
+
perimeter = cv2.arcLength(contours[0], True) if contours else 1.0
|
| 756 |
+
circularity = (4 * np.pi * area_px / (perimeter ** 2 + eps)) if perimeter > 0 else 0.0
|
| 757 |
+
|
| 758 |
+
ys_cell = grid_y[cell_mask].astype(np.float32)
|
| 759 |
+
xs_cell = grid_x[cell_mask].astype(np.float32)
|
| 760 |
+
centroid_y = ys_cell.mean()
|
| 761 |
+
centroid_x = xs_cell.mean()
|
| 762 |
+
|
| 763 |
+
cell_radius = np.sqrt(area_px / np.pi) + eps
|
| 764 |
+
dist_norm = np.sqrt((xs_cell - centroid_x)**2 + (ys_cell - centroid_y)**2) / cell_radius
|
| 765 |
+
|
| 766 |
+
v_all = hsv[:, :, 2][cell_mask]
|
| 767 |
+
|
| 768 |
+
# Tight inner core (15% radius) β captures specular highlight spot only
|
| 769 |
+
inner_mask = dist_norm < 0.15
|
| 770 |
+
# Membrane ring zone (20-60%) β dark navy ring on live cells
|
| 771 |
+
ring_mask = (dist_norm >= 0.20) & (dist_norm <= 0.60)
|
| 772 |
+
# Outer zone (>60%) β denominator for centre ratio
|
| 773 |
+
outer_mask = dist_norm > 0.60
|
| 774 |
+
|
| 775 |
+
inner_brightness = float(v_all[inner_mask].mean()) if inner_mask.any() else float(v.mean())
|
| 776 |
+
ring_brightness = float(v_all[ring_mask].mean()) if ring_mask.any() else float(v.mean())
|
| 777 |
+
outer_brightness = float(v_all[outer_mask].mean()) if outer_mask.any() else float(v.mean())
|
| 778 |
+
|
| 779 |
+
# Peak V β specular spot is just a few pixels so mean dilutes it
|
| 780 |
+
peak_brightness = float(v_all.max())
|
| 781 |
+
|
| 782 |
+
# Fraction of cell pixels with V > 200 (specular highlight region)
|
| 783 |
+
bright_spot_fraction = float((v_all > 200).sum()) / (len(v_all) + eps)
|
| 784 |
+
|
| 785 |
+
# Ring darkness: ratio of ring zone to outer zone brightness
|
| 786 |
+
# Live: ring << outer (dark membrane ring) -> ratio < 1
|
| 787 |
+
# Dead: uniform blob -> ratio ~ 1
|
| 788 |
+
ring_darkness = ring_brightness / (outer_brightness + eps)
|
| 789 |
+
|
| 790 |
+
centre_periphery_ratio = inner_brightness / (outer_brightness + eps)
|
| 791 |
+
|
| 792 |
+
brightness_std_normalised = float(v.std()) / (float(v.mean()) + eps)
|
| 793 |
+
|
| 794 |
+
rows.append({
|
| 795 |
+
"cell_id": int(cid),
|
| 796 |
+
"mean_r": float(r.mean()),
|
| 797 |
+
"mean_g": float(g.mean()),
|
| 798 |
+
"mean_b": float(b.mean()),
|
| 799 |
+
"std_r": float(r.std()),
|
| 800 |
+
"std_g": float(g.std()),
|
| 801 |
+
"std_b": float(b.std()),
|
| 802 |
+
"mean_h": float(h.mean()),
|
| 803 |
+
"mean_s": float(s.mean()),
|
| 804 |
+
"mean_v": float(v.mean()),
|
| 805 |
+
"std_s": float(s.std()),
|
| 806 |
+
"std_v": float(v.std()),
|
| 807 |
+
"blue_red_ratio": round(blue_red_ratio, 5),
|
| 808 |
+
"blue_green_ratio": round(blue_green_ratio, 5),
|
| 809 |
+
"rg_ratio": round(rg_ratio, 5),
|
| 810 |
+
"area_px": area_px,
|
| 811 |
+
"circularity": round(float(circularity), 5),
|
| 812 |
+
"inner_brightness": round(inner_brightness, 3),
|
| 813 |
+
"peak_brightness": round(peak_brightness, 3),
|
| 814 |
+
"bright_spot_fraction": round(bright_spot_fraction, 6),
|
| 815 |
+
"ring_darkness": round(ring_darkness, 5),
|
| 816 |
+
"centre_periphery_ratio": round(centre_periphery_ratio, 5),
|
| 817 |
+
"brightness_std_normalised": round(brightness_std_normalised, 5),
|
| 818 |
+
})
|
| 819 |
+
|
| 820 |
+
return rows
|
| 821 |
+
|
| 822 |
+
def attach_viability_labels(cell_features, masks, image_np, label_map=None):
|
| 823 |
+
"""
|
| 824 |
+
Attach model predictions (from label_map) to each feature dict.
|
| 825 |
+
label_map: {cell_id: 0=live, 1=dead} from classify_cells_by_model.
|
| 826 |
+
If label_map is None, defaults all labels to 0 (live).
|
| 827 |
+
"""
|
| 828 |
+
if not cell_features:
|
| 829 |
+
return []
|
| 830 |
+
labelled = []
|
| 831 |
+
for feat in cell_features:
|
| 832 |
+
row = dict(feat)
|
| 833 |
+
cid = int(feat["cell_id"])
|
| 834 |
+
row["label"] = int(label_map.get(cid, 0)) if label_map else 0
|
| 835 |
+
row["corrected"] = False
|
| 836 |
+
labelled.append(row)
|
| 837 |
+
return labelled
|
| 838 |
+
|
| 839 |
+
|
| 840 |
+
def export_cell_data_csv(cell_data):
|
| 841 |
+
"""Write cell_data list-of-dicts to a temp CSV and return the file path."""
|
| 842 |
+
if not cell_data:
|
| 843 |
+
return None
|
| 844 |
+
tmp = tempfile.NamedTemporaryFile(
|
| 845 |
+
mode="w", suffix=".csv", delete=False, newline=""
|
| 846 |
+
)
|
| 847 |
+
# Union of all keys across rows so any late-added keys (e.g. "corrected") are included
|
| 848 |
+
fieldnames = list(dict.fromkeys(k for row in cell_data for k in row.keys()))
|
| 849 |
+
writer = csv.DictWriter(tmp, fieldnames=fieldnames, extrasaction="ignore")
|
| 850 |
+
writer.writeheader()
|
| 851 |
+
writer.writerows(cell_data)
|
| 852 |
+
tmp.close()
|
| 853 |
+
return tmp.name
|
| 854 |
+
|
| 855 |
+
|
| 856 |
+
def prepare_export(stored_masks, stored_image, threshold_bias):
|
| 857 |
+
"""
|
| 858 |
+
Called by the Export button. Unpacks state, extracts features,
|
| 859 |
+
attaches labels, writes CSV, returns (path, status_message).
|
| 860 |
+
"""
|
| 861 |
+
if stored_masks is None or stored_image is None:
|
| 862 |
+
return None, "Run segmentation first before exporting."
|
| 863 |
+
|
| 864 |
+
masks = unpack_array(stored_masks)
|
| 865 |
+
image_np = unpack_array(stored_image)
|
| 866 |
+
|
| 867 |
+
features = extract_cell_features(image_np, masks)
|
| 868 |
+
if not features:
|
| 869 |
+
return None, "No cells found to export."
|
| 870 |
+
|
| 871 |
+
labelled = attach_viability_labels(features, masks, image_np, threshold_bias)
|
| 872 |
+
path = export_cell_data_csv(labelled)
|
| 873 |
+
|
| 874 |
+
n = len(labelled)
|
| 875 |
+
dead = sum(1 for r in labelled if r["label"] == 1)
|
| 876 |
+
alive = n - dead
|
| 877 |
+
msg = (f"Exported {n} cells ({alive} live, {dead} dead) β "
|
| 878 |
+
f"threshold bias={threshold_bias:+d}.\n"
|
| 879 |
+
f"Columns: {', '.join(list(labelled[0].keys())[:6])}β¦ "
|
| 880 |
+
f"({len(labelled[0])} total).")
|
| 881 |
+
return path, msg
|
| 882 |
+
|
| 883 |
+
|
| 884 |
+
|
| 885 |
+
# Tab builder
|
| 886 |
+
|
| 887 |
+
def draw_polygon_overlay(image_pil, points):
|
| 888 |
+
"""
|
| 889 |
+
Draw numbered vertex dots and polygon edges onto a copy of image_pil.
|
| 890 |
+
points: list of (x, y) tuples in original image pixel space.
|
| 891 |
+
Returns a new PIL image.
|
| 892 |
+
"""
|
| 893 |
+
img = image_pil.copy().convert("RGBA")
|
| 894 |
+
overlay = Image.new("RGBA", img.size, (0, 0, 0, 0))
|
| 895 |
+
draw = ImageDraw.Draw(overlay)
|
| 896 |
+
|
| 897 |
+
if len(points) >= 2:
|
| 898 |
+
# Draw edges
|
| 899 |
+
for i in range(len(points) - 1):
|
| 900 |
+
draw.line([points[i], points[i + 1]], fill=(74, 170, 255, 220), width=3)
|
| 901 |
+
if len(points) == 4:
|
| 902 |
+
draw.line([points[-1], points[0]], fill=(74, 170, 255, 220), width=3)
|
| 903 |
+
# Semi-transparent fill
|
| 904 |
+
draw.polygon(points, fill=(74, 170, 255, 50))
|
| 905 |
+
|
| 906 |
+
# Draw vertex dots + numbers
|
| 907 |
+
r = max(8, min(img.width, img.height) // 60)
|
| 908 |
+
for i, (x, y) in enumerate(points):
|
| 909 |
+
draw.ellipse([x - r, y - r, x + r, y + r],
|
| 910 |
+
fill=(74, 170, 255, 255), outline=(255, 255, 255, 255))
|
| 911 |
+
draw.text((x, y), str(i + 1), fill=(255, 255, 255, 255), anchor="mm")
|
| 912 |
+
|
| 913 |
+
combined = Image.alpha_composite(img, overlay)
|
| 914 |
+
return combined.convert("RGB")
|
| 915 |
+
|
| 916 |
+
|
| 917 |
+
def add_crop_point(image_pil, points, evt: gr.SelectData):
|
| 918 |
+
"""
|
| 919 |
+
Called by gr.Image .select(). Appends the clicked coordinate,
|
| 920 |
+
redraws the overlay, returns (updated_image, updated_points).
|
| 921 |
+
Ignores clicks once 4 points are set.
|
| 922 |
+
"""
|
| 923 |
+
if image_pil is None:
|
| 924 |
+
return image_pil, points
|
| 925 |
+
if points is None:
|
| 926 |
+
points = []
|
| 927 |
+
if len(points) >= 4:
|
| 928 |
+
return draw_polygon_overlay(image_pil, points), points
|
| 929 |
+
|
| 930 |
+
x, y = int(evt.index[0]), int(evt.index[1])
|
| 931 |
+
new_points = points + [(x, y)]
|
| 932 |
+
return draw_polygon_overlay(image_pil, new_points), new_points
|
| 933 |
+
|
| 934 |
+
|
| 935 |
+
def clear_crop_points(image_pil):
|
| 936 |
+
"""Reset polygon β return original image with no overlay and empty points."""
|
| 937 |
+
return image_pil, []
|
| 938 |
+
|
| 939 |
+
|
| 940 |
+
|
| 941 |
+
|
| 942 |
+
|
| 943 |
+
# Label correction grid
|
| 944 |
+
|
| 945 |
+
THUMB_SIZE = 80
|
| 946 |
+
GRID_COLS = 10
|
| 947 |
+
BORDER = 4
|
| 948 |
+
LABEL_H = 16
|
| 949 |
+
|
| 950 |
+
def _crop_cell_thumb(image_np, masks, cid):
|
| 951 |
+
"""
|
| 952 |
+
Return a tight square crop of the cell, padded to THUMB_SIZE Γ THUMB_SIZE.
|
| 953 |
+
"""
|
| 954 |
+
ys, xs = np.where(masks == cid)
|
| 955 |
+
if len(ys) == 0:
|
| 956 |
+
return Image.fromarray(np.zeros((THUMB_SIZE, THUMB_SIZE, 3), dtype=np.uint8))
|
| 957 |
+
|
| 958 |
+
y0, y1 = ys.min(), ys.max() + 1
|
| 959 |
+
x0, x1 = xs.min(), xs.max() + 1
|
| 960 |
+
|
| 961 |
+
# add a small context border around the tight bounding box
|
| 962 |
+
pad = max(4, int(max(y1 - y0, x1 - x0) * 0.15))
|
| 963 |
+
h, w = image_np.shape[:2]
|
| 964 |
+
y0c = max(0, y0 - pad)
|
| 965 |
+
y1c = min(h, y1 + pad)
|
| 966 |
+
x0c = max(0, x0 - pad)
|
| 967 |
+
x1c = min(w, x1 + pad)
|
| 968 |
+
|
| 969 |
+
crop = image_np[y0c:y1c, x0c:x1c].copy()
|
| 970 |
+
|
| 971 |
+
# dim pixels that don't belong to this cell
|
| 972 |
+
dim_mask = (masks[y0c:y1c, x0c:x1c] != cid)
|
| 973 |
+
crop[dim_mask] = (crop[dim_mask] * 0.3).astype(np.uint8)
|
| 974 |
+
|
| 975 |
+
pil = Image.fromarray(crop).resize((THUMB_SIZE, THUMB_SIZE), Image.LANCZOS)
|
| 976 |
+
return pil
|
| 977 |
+
|
| 978 |
+
|
| 979 |
+
def build_correction_grid(image_np, masks, labelled_features, raw_image_np=None):
|
| 980 |
+
|
| 981 |
+
if not labelled_features:
|
| 982 |
+
placeholder = Image.fromarray(
|
| 983 |
+
np.zeros((THUMB_SIZE, THUMB_SIZE, 3), dtype=np.uint8)
|
| 984 |
+
)
|
| 985 |
+
return placeholder
|
| 986 |
+
|
| 987 |
+
thumb_src = raw_image_np if raw_image_np is not None else image_np
|
| 988 |
+
|
| 989 |
+
n = len(labelled_features)
|
| 990 |
+
n_cols = GRID_COLS
|
| 991 |
+
n_rows = (n + n_cols - 1) // n_cols
|
| 992 |
+
|
| 993 |
+
cell_h = THUMB_SIZE + 2 * BORDER + LABEL_H
|
| 994 |
+
cell_w = THUMB_SIZE + 2 * BORDER
|
| 995 |
+
|
| 996 |
+
grid_w = n_cols * cell_w
|
| 997 |
+
grid_h = n_rows * cell_h
|
| 998 |
+
|
| 999 |
+
grid = Image.new("RGB", (grid_w, grid_h), (30, 30, 30))
|
| 1000 |
+
draw = ImageDraw.Draw(grid)
|
| 1001 |
+
|
| 1002 |
+
for idx, feat in enumerate(labelled_features):
|
| 1003 |
+
cid = feat["cell_id"]
|
| 1004 |
+
label = feat["label"] # 0=live, 1=dead (may have been corrected)
|
| 1005 |
+
color = (220, 50, 50) if label == 1 else (50, 200, 80)
|
| 1006 |
+
|
| 1007 |
+
thumb = _crop_cell_thumb(thumb_src, masks, cid)
|
| 1008 |
+
|
| 1009 |
+
col = idx % n_cols
|
| 1010 |
+
row = idx // n_cols
|
| 1011 |
+
x0 = col * cell_w
|
| 1012 |
+
y0 = row * cell_h
|
| 1013 |
+
|
| 1014 |
+
# coloured border rectangle
|
| 1015 |
+
draw.rectangle([x0, y0, x0 + cell_w - 1, y0 + cell_h - 1], outline=color, width=BORDER)
|
| 1016 |
+
|
| 1017 |
+
# paste thumbnail inside border
|
| 1018 |
+
grid.paste(thumb, (x0 + BORDER, y0 + BORDER))
|
| 1019 |
+
|
| 1020 |
+
# small cell-id label strip
|
| 1021 |
+
strip_y = y0 + BORDER + THUMB_SIZE
|
| 1022 |
+
draw.rectangle([x0, strip_y, x0 + cell_w - 1, y0 + cell_h - 1],
|
| 1023 |
+
fill=(20, 20, 20))
|
| 1024 |
+
draw.text((x0 + BORDER + 2, strip_y + 1),
|
| 1025 |
+
f"#{cid} {'D' if label == 1 else 'L'}",
|
| 1026 |
+
fill=color)
|
| 1027 |
+
|
| 1028 |
+
return grid
|
| 1029 |
+
|
| 1030 |
+
|
| 1031 |
+
def toggle_cell_label(labelled_features, image_np, masks, raw_image_np, evt: gr.SelectData):
|
| 1032 |
+
"""
|
| 1033 |
+
Called when user taps the correction grid image.
|
| 1034 |
+
Maps the tap pixel coordinate back to which thumbnail was tapped,
|
| 1035 |
+
flips that cell's label, rebuilds and returns the updated grid.
|
| 1036 |
+
"""
|
| 1037 |
+
if not labelled_features or image_np is None:
|
| 1038 |
+
return build_correction_grid(image_np, masks, labelled_features), labelled_features
|
| 1039 |
+
|
| 1040 |
+
cell_w = THUMB_SIZE + 2 * BORDER
|
| 1041 |
+
cell_h = THUMB_SIZE + 2 * BORDER + LABEL_H
|
| 1042 |
+
|
| 1043 |
+
px, py = int(evt.index[0]), int(evt.index[1])
|
| 1044 |
+
col = px // cell_w
|
| 1045 |
+
row = py // cell_h
|
| 1046 |
+
idx = row * GRID_COLS + col
|
| 1047 |
+
|
| 1048 |
+
if idx < 0 or idx >= len(labelled_features):
|
| 1049 |
+
return build_correction_grid(image_np, masks, labelled_features, raw_image_np), labelled_features
|
| 1050 |
+
|
| 1051 |
+
# Flip the label
|
| 1052 |
+
updated = list(labelled_features) # shallow copy of list
|
| 1053 |
+
cell = dict(updated[idx]) # copy the dict so we don't mutate in place
|
| 1054 |
+
cell["label"] = 1 - cell["label"] # 0β1 or 1β0
|
| 1055 |
+
cell["corrected"] = True
|
| 1056 |
+
updated[idx] = cell
|
| 1057 |
+
|
| 1058 |
+
grid = build_correction_grid(image_np, masks, updated, raw_image_np)
|
| 1059 |
+
n_corrected = sum(1 for f in updated if f.get("corrected"))
|
| 1060 |
+
return grid, updated, f"Tapped cell #{cell['cell_id']} β {'Dead' if cell['label']==1 else 'Live'}. {n_corrected} correction(s) total."
|
| 1061 |
+
|
| 1062 |
+
|
| 1063 |
+
def prepare_export_corrected(stored_masks, stored_image, labelled_features, label_map):
|
| 1064 |
+
"""Export CSV using labelled_features with any manual corrections applied."""
|
| 1065 |
+
if stored_masks is None or stored_image is None:
|
| 1066 |
+
return None, "Run segmentation first before exporting."
|
| 1067 |
+
masks = unpack_array(stored_masks)
|
| 1068 |
+
image_np = unpack_array(stored_image)
|
| 1069 |
+
if not labelled_features:
|
| 1070 |
+
features = extract_cell_features(image_np, masks)
|
| 1071 |
+
labelled_features = attach_viability_labels(features, masks, image_np, label_map)
|
| 1072 |
+
if not labelled_features:
|
| 1073 |
+
return None, "No cells found to export."
|
| 1074 |
+
path = export_cell_data_csv(labelled_features)
|
| 1075 |
+
n = len(labelled_features)
|
| 1076 |
+
dead = sum(1 for r in labelled_features if r["label"] == 1)
|
| 1077 |
+
alive = n - dead
|
| 1078 |
+
corrected = sum(1 for r in labelled_features if r.get("corrected"))
|
| 1079 |
+
msg = (f"Exported {n} cells ({alive} live, {dead} dead). "
|
| 1080 |
+
f"{corrected} label(s) manually corrected.")
|
| 1081 |
+
return path, msg
|
| 1082 |
+
|
| 1083 |
+
def build_tab(tab_index, masks_state, image_state, result_state):
|
| 1084 |
+
with gr.Tab(f"Tab {tab_index}"):
|
| 1085 |
+
gr.Markdown("Run segmentation")
|
| 1086 |
+
|
| 1087 |
+
# Per-tab state: list of (x,y) crop polygon points
|
| 1088 |
+
crop_points_state = gr.State(value=[])
|
| 1089 |
+
# Clean copy of the uploaded image (no polygon drawn on it)
|
| 1090 |
+
base_image_state = gr.State(value=None)
|
| 1091 |
+
#raw image state
|
| 1092 |
+
raw_image_state = gr.State(value=None)
|
| 1093 |
+
|
| 1094 |
+
with gr.Row():
|
| 1095 |
+
with gr.Column():
|
| 1096 |
+
img_input = gr.Image(
|
| 1097 |
+
type="pil",
|
| 1098 |
+
label="Upload image",
|
| 1099 |
+
image_mode="RGB",
|
| 1100 |
+
height=512
|
| 1101 |
+
)
|
| 1102 |
+
|
| 1103 |
+
gr.Markdown(
|
| 1104 |
+
"### Crop region (optional)\n"
|
| 1105 |
+
"Click/tap up to **4 points** on the image below to define the region "
|
| 1106 |
+
"to segment. The polygon will be drawn as you click. "
|
| 1107 |
+
"Leave empty to segment the full image."
|
| 1108 |
+
)
|
| 1109 |
+
|
| 1110 |
+
crop_display = gr.Image(
|
| 1111 |
+
type="pil",
|
| 1112 |
+
label="Click to set crop vertices (up to 4)",
|
| 1113 |
+
interactive=True,
|
| 1114 |
+
height=400,
|
| 1115 |
+
)
|
| 1116 |
+
|
| 1117 |
+
crop_status = gr.Markdown("*Upload an image to enable cropping*")
|
| 1118 |
+
|
| 1119 |
+
clear_crop_btn = gr.Button("β Clear crop points", size="sm")
|
| 1120 |
+
|
| 1121 |
+
model_dropdown = gr.Dropdown(
|
| 1122 |
+
choices=list(MODEL_OPTIONS.keys()),
|
| 1123 |
+
label="Select Model",
|
| 1124 |
+
value="Hemocytometer Model"
|
| 1125 |
+
)
|
| 1126 |
+
|
| 1127 |
+
gr.Markdown("### Size Filters")
|
| 1128 |
+
|
| 1129 |
+
use_min_filter = gr.Checkbox(
|
| 1130 |
+
label="Enable minimum size filter",
|
| 1131 |
+
value=False,
|
| 1132 |
+
info="Remove objects smaller than the threshold below"
|
| 1133 |
+
)
|
| 1134 |
+
min_size_slider = gr.Slider(
|
| 1135 |
+
minimum=0,
|
| 1136 |
+
maximum=500,
|
| 1137 |
+
value=0,
|
| 1138 |
+
step=10,
|
| 1139 |
+
label="Minimum Cell Size (pixels)",
|
| 1140 |
+
)
|
| 1141 |
+
min_size_recommendation = gr.Markdown(
|
| 1142 |
+
value="*Run segmentation to see recommended minimum*",
|
| 1143 |
+
)
|
| 1144 |
+
|
| 1145 |
+
use_max_filter = gr.Checkbox(
|
| 1146 |
+
label="Enable maximum size filter",
|
| 1147 |
+
value=False,
|
| 1148 |
+
info="Remove objects larger than the threshold below"
|
| 1149 |
+
)
|
| 1150 |
+
max_size_slider = gr.Slider(
|
| 1151 |
+
minimum=0,
|
| 1152 |
+
maximum=10000,
|
| 1153 |
+
value=10000,
|
| 1154 |
+
step=10,
|
| 1155 |
+
label="Maximum Cell Size (pixels)",
|
| 1156 |
+
)
|
| 1157 |
+
|
| 1158 |
+
gr.Markdown("### Stereological Counting")
|
| 1159 |
+
use_stereo = gr.Checkbox(
|
| 1160 |
+
label="Enable Stereological Counting",
|
| 1161 |
+
value=False,
|
| 1162 |
+
info="Use unbiased stereological rules for cell counting"
|
| 1163 |
+
)
|
| 1164 |
+
|
| 1165 |
+
with gr.Group(visible=False) as stereo_controls:
|
| 1166 |
+
gr.Markdown("""
|
| 1167 |
+
**Stereological Counting Rules:**
|
| 1168 |
+
- Cells touching LEFT or TOP exclusion zones are EXCLUDED
|
| 1169 |
+
- Cells touching RIGHT or BOTTOM edges are INCLUDED
|
| 1170 |
+
- This provides unbiased counting for quantification
|
| 1171 |
+
""")
|
| 1172 |
+
|
| 1173 |
+
excl_preview = gr.Image(
|
| 1174 |
+
type="pil",
|
| 1175 |
+
label="Exclusion Zone Preview (Red = Excluded)",
|
| 1176 |
+
height=500
|
| 1177 |
+
)
|
| 1178 |
+
|
| 1179 |
+
left_excl = gr.Slider(
|
| 1180 |
+
minimum=0,
|
| 1181 |
+
maximum=50,
|
| 1182 |
+
value=10,
|
| 1183 |
+
step=1,
|
| 1184 |
+
label="Left Exclusion Width (%)",
|
| 1185 |
+
info="Width of left exclusion zone"
|
| 1186 |
+
)
|
| 1187 |
+
|
| 1188 |
+
top_excl = gr.Slider(
|
| 1189 |
+
minimum=0,
|
| 1190 |
+
maximum=50,
|
| 1191 |
+
value=10,
|
| 1192 |
+
step=1,
|
| 1193 |
+
label="Top Exclusion Width (%)",
|
| 1194 |
+
info="Width of top exclusion zone"
|
| 1195 |
+
)
|
| 1196 |
+
|
| 1197 |
+
segment_btn = gr.Button("π¬ Run Segmentation", variant="primary", size="lg")
|
| 1198 |
+
|
| 1199 |
+
with gr.Column():
|
| 1200 |
+
cell_count_out = gr.Number(label="Total Cells Detected", precision=0)
|
| 1201 |
+
confluency_out = gr.Number(label="Confluency (%)", precision=1)
|
| 1202 |
+
overlay_out = gr.Image(type="pil", label="Segmentation Result")
|
| 1203 |
+
info_out = gr.Textbox(label="Processing Info", lines=4)
|
| 1204 |
+
|
| 1205 |
+
with gr.Group(visible=False) as viability_section:
|
| 1206 |
+
gr.Markdown("### Viability Assessment (Trypan Blue)")
|
| 1207 |
+
|
| 1208 |
+
viab_run_btn = gr.Button("Run Viability Analysis", variant="primary")
|
| 1209 |
+
|
| 1210 |
+
with gr.Row():
|
| 1211 |
+
live_count_out = gr.Number(label="Live Cells (Green)", precision=0)
|
| 1212 |
+
dead_count_out = gr.Number(label="Dead Cells (Red)", precision=0)
|
| 1213 |
+
|
| 1214 |
+
viab_overlay = gr.Image(type="pil", label="Viability (Green=Live Β· Red=Dead)")
|
| 1215 |
+
viab_percent_out = gr.Number(label="Viability (%)", precision=1)
|
| 1216 |
+
viab_info = gr.Textbox(label="Analysis Results", lines=4)
|
| 1217 |
+
|
| 1218 |
+
gr.Markdown("### Label Correction & Export")
|
| 1219 |
+
gr.Markdown(
|
| 1220 |
+
"After running viability, click **Build correction grid** to review every cell. "
|
| 1221 |
+
"**Green border = Live, Red border = Dead** (model predictions). "
|
| 1222 |
+
"Tap any thumbnail to flip its label β the counts and overlay update instantly. "
|
| 1223 |
+
"Export the corrected CSV for retraining."
|
| 1224 |
+
)
|
| 1225 |
+
|
| 1226 |
+
build_grid_btn = gr.Button("π² Build correction grid", variant="secondary")
|
| 1227 |
+
labelled_state = gr.State(value=[])
|
| 1228 |
+
label_map_state = gr.State(value={})
|
| 1229 |
+
|
| 1230 |
+
correction_grid = gr.Image(
|
| 1231 |
+
type="pil",
|
| 1232 |
+
label="Tap a cell to flip its label (green=live Β· red=dead)",
|
| 1233 |
+
interactive=True,
|
| 1234 |
+
visible=False,
|
| 1235 |
+
)
|
| 1236 |
+
correction_status = gr.Markdown(visible=False)
|
| 1237 |
+
|
| 1238 |
+
with gr.Row():
|
| 1239 |
+
export_btn = gr.Button("β¬οΈ Export corrected CSV", variant="secondary")
|
| 1240 |
+
export_info = gr.Textbox(label="Export status", lines=2, interactive=False)
|
| 1241 |
+
export_file = gr.File(label="Download CSV", visible=False)
|
| 1242 |
+
|
| 1243 |
+
# ---- Event handlers ------------------------------------------------
|
| 1244 |
+
|
| 1245 |
+
use_stereo.change(
|
| 1246 |
+
fn=toggle_stereological_mode,
|
| 1247 |
+
inputs=[use_stereo],
|
| 1248 |
+
outputs=[stereo_controls]
|
| 1249 |
+
)
|
| 1250 |
+
|
| 1251 |
+
def on_image_upload(img):
|
| 1252 |
+
if img is None:
|
| 1253 |
+
return None, None, "*Upload an image to enable cropping*"
|
| 1254 |
+
return img, img, "*Image loaded β click up to 4 points to define crop region*"
|
| 1255 |
+
|
| 1256 |
+
img_input.change(
|
| 1257 |
+
fn=on_image_upload,
|
| 1258 |
+
inputs=[img_input],
|
| 1259 |
+
outputs=[crop_display, base_image_state, crop_status]
|
| 1260 |
+
).then(fn=lambda: [], outputs=[crop_points_state])
|
| 1261 |
+
|
| 1262 |
+
img_input.change(fn=update_exclusion_preview,
|
| 1263 |
+
inputs=[img_input, left_excl, top_excl], outputs=[excl_preview])
|
| 1264 |
+
left_excl.change(fn=update_exclusion_preview,
|
| 1265 |
+
inputs=[img_input, left_excl, top_excl], outputs=[excl_preview])
|
| 1266 |
+
top_excl.change(fn=update_exclusion_preview,
|
| 1267 |
+
inputs=[img_input, left_excl, top_excl], outputs=[excl_preview])
|
| 1268 |
+
|
| 1269 |
+
def on_crop_click(base_img, points, evt: gr.SelectData):
|
| 1270 |
+
updated_img, updated_pts = add_crop_point(base_img, points, evt)
|
| 1271 |
+
n = len(updated_pts)
|
| 1272 |
+
status = (f"*{n} / 4 points set β keep clicking*" if n < 4
|
| 1273 |
+
else "*4 points set β β click **β Clear** to redo, or run segmentation*")
|
| 1274 |
+
return updated_img, updated_pts, status
|
| 1275 |
+
|
| 1276 |
+
crop_display.select(fn=on_crop_click,
|
| 1277 |
+
inputs=[base_image_state, crop_points_state],
|
| 1278 |
+
outputs=[crop_display, crop_points_state, crop_status])
|
| 1279 |
+
|
| 1280 |
+
def on_clear_crop(base_img):
|
| 1281 |
+
img, pts = clear_crop_points(base_img)
|
| 1282 |
+
return img, pts, "*Points cleared β click to set new vertices*"
|
| 1283 |
+
|
| 1284 |
+
clear_crop_btn.click(fn=on_clear_crop,
|
| 1285 |
+
inputs=[base_image_state],
|
| 1286 |
+
outputs=[crop_display, crop_points_state, crop_status])
|
| 1287 |
+
|
| 1288 |
+
segment_btn.click(
|
| 1289 |
+
fn=run_segmentation,
|
| 1290 |
+
inputs=[img_input, model_dropdown, min_size_slider, max_size_slider,
|
| 1291 |
+
use_min_filter, use_max_filter,
|
| 1292 |
+
use_stereo, left_excl, top_excl, crop_points_state],
|
| 1293 |
+
outputs=[cell_count_out, overlay_out, info_out, viability_section,
|
| 1294 |
+
masks_state, image_state, confluency_out, min_size_recommendation, raw_image_state]
|
| 1295 |
+
)
|
| 1296 |
+
|
| 1297 |
+
# ---- Run Viability button -------------------------------------------
|
| 1298 |
+
def on_run_viability(stored_masks, stored_image):
|
| 1299 |
+
overlay, alive, dead, viab_pct, info, label_map = run_viability(stored_masks, stored_image)
|
| 1300 |
+
return overlay, alive, dead, viab_pct, info, label_map
|
| 1301 |
+
|
| 1302 |
+
viab_run_btn.click(
|
| 1303 |
+
fn=on_run_viability,
|
| 1304 |
+
inputs=[masks_state, image_state],
|
| 1305 |
+
outputs=[viab_overlay, live_count_out, dead_count_out,
|
| 1306 |
+
viab_percent_out, viab_info, label_map_state]
|
| 1307 |
+
).then(
|
| 1308 |
+
fn=save_tab_result,
|
| 1309 |
+
inputs=[cell_count_out, confluency_out, viab_percent_out],
|
| 1310 |
+
outputs=[result_state]
|
| 1311 |
+
)
|
| 1312 |
+
|
| 1313 |
+
# ---- Build correction grid -----------------------------------------
|
| 1314 |
+
def on_build_grid(stored_masks, stored_image, label_map, stored_raw_image):
|
| 1315 |
+
if stored_masks is None or stored_image is None or not label_map:
|
| 1316 |
+
return (gr.update(visible=False), [],
|
| 1317 |
+
gr.update(value="*Run viability analysis first.*", visible=True))
|
| 1318 |
+
masks = unpack_array(stored_masks)
|
| 1319 |
+
image_np = unpack_array(stored_image)
|
| 1320 |
+
raw_image_np = unpack_array(stored_raw_image) if stored_raw_image is not None else None
|
| 1321 |
+
features = extract_cell_features(image_np, masks)
|
| 1322 |
+
labelled = attach_viability_labels(features, masks, image_np, label_map)
|
| 1323 |
+
if not labelled:
|
| 1324 |
+
return (gr.update(visible=False), [],
|
| 1325 |
+
gr.update(value="*No cells found.*", visible=True))
|
| 1326 |
+
grid = build_correction_grid(image_np, masks, labelled, raw_image_np)
|
| 1327 |
+
n = len(labelled)
|
| 1328 |
+
dead = sum(1 for r in labelled if r["label"] == 1)
|
| 1329 |
+
msg = (f"*{n} cells β {n-dead} live (green), {dead} dead (red). "
|
| 1330 |
+
f"Tap any thumbnail to flip its label.*")
|
| 1331 |
+
return gr.update(value=grid, visible=True), labelled, gr.update(value=msg, visible=True)
|
| 1332 |
+
|
| 1333 |
+
build_grid_btn.click(
|
| 1334 |
+
fn=on_build_grid,
|
| 1335 |
+
inputs=[masks_state, image_state, label_map_state, raw_image_state],
|
| 1336 |
+
outputs=[correction_grid, labelled_state, correction_status]
|
| 1337 |
+
)
|
| 1338 |
+
|
| 1339 |
+
# ---- Grid tap β flip label, update overlay + counts ----------------
|
| 1340 |
+
def on_grid_tap(labelled, stored_masks, stored_image, stored_raw_image, evt: gr.SelectData):
|
| 1341 |
+
if not labelled or stored_masks is None:
|
| 1342 |
+
return None, labelled, "", 0, 0, 0.0, None, {}
|
| 1343 |
+
masks = unpack_array(stored_masks)
|
| 1344 |
+
image_np = unpack_array(stored_image)
|
| 1345 |
+
raw_image_np = unpack_array(stored_raw_image) if stored_raw_image is not None else None
|
| 1346 |
+
grid, updated, msg = toggle_cell_label(labelled, image_np, masks, raw_image_np, evt)
|
| 1347 |
+
|
| 1348 |
+
# Rebuild label_map from corrected labelled list
|
| 1349 |
+
new_label_map = {int(f["cell_id"]): int(f["label"]) for f in updated}
|
| 1350 |
+
overlay_np = draw_viability_overlay(image_np, masks, new_label_map)
|
| 1351 |
+
dead = sum(1 for f in updated if f["label"] == 1)
|
| 1352 |
+
alive = len(updated) - dead
|
| 1353 |
+
total = alive + dead
|
| 1354 |
+
viab_pct = (alive / total * 100) if total > 0 else 0.0
|
| 1355 |
+
|
| 1356 |
+
return (grid, updated, f"*{msg}*",
|
| 1357 |
+
alive, dead, viab_pct,
|
| 1358 |
+
Image.fromarray(overlay_np), new_label_map)
|
| 1359 |
+
|
| 1360 |
+
correction_grid.select(
|
| 1361 |
+
fn=on_grid_tap,
|
| 1362 |
+
inputs=[labelled_state, masks_state, image_state, raw_image_state],
|
| 1363 |
+
outputs=[correction_grid, labelled_state, correction_status,
|
| 1364 |
+
live_count_out, dead_count_out, viab_percent_out,
|
| 1365 |
+
viab_overlay, label_map_state]
|
| 1366 |
+
)
|
| 1367 |
+
|
| 1368 |
+
# ---- Export --------------------------------------------------------
|
| 1369 |
+
def on_export(stored_masks, stored_image, labelled, label_map):
|
| 1370 |
+
path, msg = prepare_export_corrected(stored_masks, stored_image, labelled, label_map)
|
| 1371 |
+
if path is None:
|
| 1372 |
+
return gr.update(visible=False), msg
|
| 1373 |
+
return gr.update(value=path, visible=True), msg
|
| 1374 |
+
|
| 1375 |
+
export_btn.click(
|
| 1376 |
+
fn=on_export,
|
| 1377 |
+
inputs=[masks_state, image_state, labelled_state, label_map_state],
|
| 1378 |
+
outputs=[export_file, export_info]
|
| 1379 |
+
)
|
| 1380 |
+
|
| 1381 |
+
|
| 1382 |
+
|
| 1383 |
+
# Gradio interface
|
| 1384 |
+
|
| 1385 |
+
with gr.Blocks(
|
| 1386 |
+
title="CellposeCellCounter",
|
| 1387 |
+
theme=gr.themes.Soft(),
|
| 1388 |
+
) as demo:
|
| 1389 |
+
gr.Markdown("# CellposeCellCounter")
|
| 1390 |
+
gr.Markdown("For accurate cell confluency, crop the image to display only desired area. Note that some image file types are not yet supported. PNG and JPEG are preferred.")
|
| 1391 |
+
|
| 1392 |
+
# Shared mask/image state (one pair per tab so tabs don't clobber each other)
|
| 1393 |
+
masks_states = [gr.State(value=None) for _ in range(4)]
|
| 1394 |
+
image_states = [gr.State(value=None) for _ in range(4)]
|
| 1395 |
+
result_states = [gr.State(value=None) for _ in range(4)]
|
| 1396 |
+
|
| 1397 |
+
# Build Tabs 1β4 with a loop
|
| 1398 |
+
for i in range(4):
|
| 1399 |
+
build_tab(i + 1, masks_states[i], image_states[i], result_states[i])
|
| 1400 |
+
|
| 1401 |
+
# -------------------------------------------------------------------------
|
| 1402 |
+
# Tab 5 β Summary
|
| 1403 |
+
# -------------------------------------------------------------------------
|
| 1404 |
+
with gr.Tab("Tab 5 β Summary"):
|
| 1405 |
+
gr.Markdown("## Average Results Across All Tabs")
|
| 1406 |
+
gr.Markdown(
|
| 1407 |
+
"Run segmentation in one or more tabs, "
|
| 1408 |
+
"then click **Refresh Summary** to see the averages."
|
| 1409 |
+
)
|
| 1410 |
+
|
| 1411 |
+
refresh_btn = gr.Button("π Refresh Summary", variant="primary", size="lg")
|
| 1412 |
+
|
| 1413 |
+
with gr.Row():
|
| 1414 |
+
avg_count_out = gr.Number(label="Avg Cell Count", precision=1)
|
| 1415 |
+
avg_conf_out = gr.Number(label="Avg Confluency (%)", precision=1)
|
| 1416 |
+
avg_viab_out = gr.Number(label="Avg Viability (%)", precision=1)
|
| 1417 |
+
|
| 1418 |
+
summary_box = gr.Textbox(label="Per-Tab Breakdown", lines=10)
|
| 1419 |
+
|
| 1420 |
+
refresh_btn.click(
|
| 1421 |
+
fn=compute_summary,
|
| 1422 |
+
inputs=result_states, # list of 4 gr.State components
|
| 1423 |
+
outputs=[avg_count_out, avg_conf_out, avg_viab_out, summary_box]
|
| 1424 |
+
)
|
| 1425 |
+
|
| 1426 |
+
|
| 1427 |
+
|
| 1428 |
+
if __name__ == "__main__":
|
| 1429 |
+
demo.launch()
|