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f680f54 | 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 | import os
import glob
import logging
import random
import cv2
import numpy as np
import matplotlib.pyplot as plt
import tifffile as tiff
from scipy.sparse import csr_matrix
from tabulate import tabulate
from tqdm import tqdm
from huggingface_hub import snapshot_download
from nellie.im_info.verifier import FileInfo, ImInfo
from nellie.segmentation.filtering import Filter
from nellie.segmentation.labelling import Label
from skimage.filters import rank, sobel
from skimage import exposure
from skimage.morphology import disk, binary_dilation
from skimage.segmentation import find_boundaries
from skimage.util import img_as_ubyte
# Logging setup
logging.basicConfig(level=logging.INFO, format="%(asctime)s | %(levelname)s | %(message)s", datefmt="%H:%M:%S")
logger = logging.getLogger(__name__)
# =====================================================================
# Preprocessing Logic
# =====================================================================
def preprocess_method_1(image_obj: np.ndarray) -> np.ndarray:
""" W2: Organelles (Red Fluorescence) -> CLAHE & Top-Hat preprocessing """
if image_obj.ndim == 3:
image_obj = image_obj[0]
p1, p99 = np.percentile(image_obj, (1, 99.9))
robust_img = exposure.rescale_intensity(image_obj, in_range=(p1, p99), out_range=(0, 255)).astype(np.uint8)
clahe = cv2.createCLAHE(clipLimit=5.0, tileGridSize=(8, 8))
clahe_img = clahe.apply(robust_img)
return clahe_img
def preprocess_method_2(img_input: np.ndarray) -> np.ndarray:
img_min, img_max = img_input.min(), img_input.max()
img_normalized = (img_input - img_min) / (img_max - img_min if img_max > img_min else 1.0)
img_8bit = img_as_ubyte(img_normalized)
nellie_input = rank.entropy(img_8bit, disk(25))
edge_map = sobel(img_normalized)
texture_features = nellie_input * (1.0 + edge_map * 5.0)
tex_min, tex_max = texture_features.min(), texture_features.max()
tex_normalized = (texture_features - tex_min) / (tex_max - tex_min if tex_max > tex_min else 1.0)
return (tex_normalized * 255).astype(np.uint8)
def preprocess_combined(img_raw: np.ndarray) -> np.ndarray:
return preprocess_method_2(preprocess_method_1(img_raw))
# =====================================================================
# Vectorized Metric Evaluation
# =====================================================================
def evaluate_predictions(pred_mask: np.ndarray, gt_mask: np.ndarray, iou_threshold: float = 0.5):
"""Vectorized instance-level matching using strict IoU >= 0.5"""
pred_mask, gt_mask = pred_mask.astype(np.int32), gt_mask.astype(np.int32)
pred_labels = np.unique(pred_mask)[np.unique(pred_mask) != 0]
gt_labels = np.unique(gt_mask)[np.unique(gt_mask) != 0]
num_pred, num_gt = len(pred_labels), len(gt_labels)
pred_bg, gt_bg = pred_mask > 0, gt_mask > 0
union_pixel = np.logical_or(pred_bg, gt_bg).sum()
global_pixel_iou = np.logical_and(pred_bg, gt_bg).sum() / union_pixel if union_pixel > 0 else 0.0
if num_gt == 0 and num_pred == 0: return global_pixel_iou, 1.0, 1.0, 1.0, num_pred, num_gt
if num_gt == 0 or num_pred == 0: return global_pixel_iou, 0.0, 0.0, 0.0, num_pred, num_gt
overlapping = gt_bg & pred_bg
if not np.any(overlapping): return global_pixel_iou, 0.0, 0.0, 0.0, num_pred, num_gt
gt_id_map = {id_: i for i, id_ in enumerate(gt_labels)}
pred_id_map = {id_: j for j, id_ in enumerate(pred_labels)}
gt_indices = np.array([gt_id_map[x] for x in gt_mask[overlapping]])
pred_indices = np.array([pred_id_map[x] for x in pred_mask[overlapping]])
intersection = csr_matrix((np.ones(len(gt_indices), dtype=np.int32), (gt_indices, pred_indices)), shape=(num_gt, num_pred)).toarray()
gt_vec = np.array([np.bincount(gt_mask.ravel())[id_] for id_ in gt_labels])[:, None]
pred_vec = np.array([np.bincount(pred_mask.ravel())[id_] for id_ in pred_labels])[None, :]
union = gt_vec + pred_vec - intersection
iou_matrix = np.divide(intersection, union, out=np.zeros_like(intersection, dtype=float), where=union != 0)
matches = iou_matrix > iou_threshold
tp = np.sum(np.any(matches, axis=0))
precision = tp / num_pred if num_pred > 0 else 0.0
recall = tp / num_gt if num_gt > 0 else 0.0
instance_f1 = (2 * precision * recall) / (precision + recall) if (precision + recall) > 0 else 0.0
return global_pixel_iou, precision, recall, instance_f1, num_pred, num_gt
# =====================================================================
# Dataset Prep & Core Pipeline
# =====================================================================
def prepare_datasets(raw_tif_files, base_dir="generated_datasets"):
dirs = {
"Raw": os.path.join(base_dir, "dataset_raw"),
"Prep 1": os.path.join(base_dir, "dataset_prep1"),
"Prep 2": os.path.join(base_dir, "dataset_prep2"),
"Prep 1->2": os.path.join(base_dir, "dataset_prep1_2")
}
for d in dirs.values(): os.makedirs(d, exist_ok=True)
for filepath in tqdm(raw_tif_files, desc="Building Datasets"):
fname = os.path.basename(filepath)
if os.path.exists(os.path.join(dirs["Prep 1->2"], fname)):
continue
img_raw = np.squeeze(tiff.imread(filepath))
tiff.imwrite(os.path.join(dirs["Raw"], fname), img_raw)
tiff.imwrite(os.path.join(dirs["Prep 1"], fname), preprocess_method_1(img_raw))
tiff.imwrite(os.path.join(dirs["Prep 2"], fname), preprocess_method_2(img_raw))
tiff.imwrite(os.path.join(dirs["Prep 1->2"], fname), preprocess_combined(img_raw))
return dirs
def run_nellie_experiment(dataset_dir, combined_data_path, run_native_filter, target_filenames):
files = [os.path.join(dataset_dir, fname) for fname in target_filenames]
results, saved_masks = [], {}
for filepath in tqdm(files, desc=f"Evaluating", leave=False):
if not os.path.exists(filepath): continue
filename = os.path.basename(filepath)
gt_mask_path = os.path.join(combined_data_path, filename.replace("_w2.TIF", "_w3_seg.npy"))
if not os.path.exists(gt_mask_path): continue
f_info = FileInfo(filepath=filepath)
f_info.find_metadata()
f_info.load_metadata()
img_array = np.squeeze(tiff.imread(filepath))
f_info.axes = "ZYX" if img_array.ndim == 3 else "YX"
f_info.good_axes, f_info.good_dims = True, True
im_info = ImInfo(file_info=f_info)
if run_native_filter:
frangi_filter = Filter(im_info=im_info, device="auto")
frangi_filter.run()
else:
prep_path = im_info.create_output_path(pipeline_path="im_preprocessed", ext=".ome.tif")
im_info.allocate_memory(output_path=prep_path, data=img_array, dtype=str(img_array.dtype))
label_processor = Label(im_info=im_info, otsu_thresh_intensity=False, min_radius_um=10.0, device="auto")
label_processor.run()
pred_mask = np.squeeze(tiff.imread(im_info.pipeline_paths["im_instance_label"]))
gt_data = np.load(gt_mask_path, allow_pickle=True)
if gt_data.ndim == 0 and isinstance(gt_data.item(), dict):
gt_data = gt_data.item().get("masks", gt_data.item())
gt_to_compare = np.squeeze(gt_data)
metrics = evaluate_predictions(pred_mask, gt_to_compare, iou_threshold=0.5)
results.append(metrics)
saved_masks[filename] = (pred_mask, metrics[4])
return results, saved_masks
# =====================================================================
# Plot Single Experiment
# =====================================================================
def plot_single_experiment_visual(sample_filename, combined_data_path, data_src_dir, raw_data_dir, config_name, pred_mask, pred_count):
logger.info(f"Generating visual plot for: {config_name}")
# --- Load Data ---
gt_data = np.load(os.path.join(combined_data_path, sample_filename.replace("_w2.TIF", "_w3_seg.npy")), allow_pickle=True)
if gt_data.ndim == 0 and isinstance(gt_data.item(), dict):
gt_data = gt_data.item().get("masks", gt_data.item())
gt_mask = np.squeeze(gt_data)
gt_count = len(np.unique(gt_mask)[np.unique(gt_mask) != 0])
feed_img = tiff.imread(os.path.join(data_src_dir, sample_filename))
raw_img = tiff.imread(os.path.join(raw_data_dir, sample_filename))
# --- Ground Truth High-Def Assets ---
gt_binary = gt_mask > 0
gt_boundaries = find_boundaries(gt_binary, mode="outer")
gt_thick_boundaries = binary_dilation(gt_boundaries, disk(1))
gt_masked_fill = np.ma.masked_where(~gt_binary, gt_binary)
gt_masked_edges = np.ma.masked_where(~gt_thick_boundaries, gt_thick_boundaries)
# --- Prediction High-Def Assets ---
pred_binary = pred_mask > 0
pred_boundaries = find_boundaries(pred_binary, mode="outer")
pred_thick_boundaries = binary_dilation(pred_boundaries, disk(1))
pred_masked_fill = np.ma.masked_where(~pred_binary, pred_binary)
pred_masked_edges = np.ma.masked_where(~pred_thick_boundaries, pred_thick_boundaries)
cmap_choice = "gray" if "Prep 1" in data_src_dir or "Raw" in data_src_dir else "magma"
fig, axes = plt.subplots(1, 3, figsize=(18, 6))
# 1. Ground Truth Overlay (on Raw Image)
axes[0].imshow(raw_img, cmap="gray")
axes[0].imshow(gt_masked_fill, cmap="summer", alpha=0.25)
axes[0].imshow(gt_masked_edges, cmap="Greens", alpha=0.9)
axes[0].set_title(f"GT Overlay on Raw (Count: {gt_count})", fontsize=12, fontweight="bold")
axes[0].axis("off")
# 2. Input Image Alone (The preprocessed image actually fed to Nellie)
axes[1].imshow(feed_img, cmap=cmap_choice)
axes[1].set_title(f"Data Fed into Nellie", fontsize=12, fontweight="bold")
axes[1].axis("off")
# 3. Predicted Mask Overlay (on Raw Image)
axes[2].imshow(raw_img, cmap="gray")
axes[2].imshow(pred_masked_fill, cmap="autumn", alpha=0.25)
axes[2].imshow(pred_masked_edges, cmap="Wistia", alpha=0.9)
axes[2].set_title(f"Predicted Mask on Raw (Count: {pred_count})", fontsize=12, fontweight="bold")
axes[2].axis("off")
plt.suptitle(f"EXPERIMENT: {config_name}\nFile: {sample_filename}", fontsize=16, fontweight="bold")
plt.tight_layout()
plt.show()
# =====================================================================
# Main Execution Flow
# =====================================================================
def main():
n_samples = 10 # Evaluate 10 images
logger.info("Downloading dataset...")
dataset_dir = snapshot_download(repo_id="champ7/celldatamag", repo_type="dataset", allow_patterns="combined_data/*")
combined_data_path = os.path.join(dataset_dir, "combined_data")
# Extract complete list and sample N files
raw_tif_files = sorted(glob.glob(os.path.join(combined_data_path, "*.TIF")))
if n_samples and n_samples < len(raw_tif_files):
random.seed(42) # Ensure the same random subset is used across runs
raw_tif_files = random.sample(raw_tif_files, n_samples)
logger.info(f"Subsampled dataset to {n_samples} random images.")
target_filenames = [os.path.basename(f) for f in raw_tif_files]
sample_filename = target_filenames[0]
dirs = prepare_datasets(raw_tif_files)
configs = [
{"name": "Prep 1 Only (Bypassing Nellie Filter)", "data_src": "Prep 1", "run_filter": False},
{"name": "Prep 2 Only (Bypassing Nellie Filter)", "data_src": "Prep 2", "run_filter": False},
{"name": "Prep 1->2 Only (Bypassing Nellie Filter)", "data_src": "Prep 1->2", "run_filter": False},
{"name": "Nellie Native Only (Raw -> Nellie Filter)", "data_src": "Raw", "run_filter": True},
{"name": "Prep 1 + Nellie Native Filter", "data_src": "Prep 1", "run_filter": True},
{"name": "Prep 2 + Nellie Native Filter", "data_src": "Prep 2", "run_filter": True},
{"name": "Prep 1->2 + Nellie Native Filter", "data_src": "Prep 1->2", "run_filter": True},
]
aggregate_table = []
headers = ["Strategy Configuration", "Total GT Cells", "Total Pred Cells", "Mean Pixel IoU", "Mean Precision", "Mean Recall", "Mean F1"]
for config in configs:
logger.info(f"Running strategy: {config['name']}")
metrics, saved_masks = run_nellie_experiment(
dataset_dir=dirs[config["data_src"]],
combined_data_path=combined_data_path,
run_native_filter=config["run_filter"],
target_filenames=target_filenames
)
ious, precs, recs, f1s = [m[0] for m in metrics], [m[1] for m in metrics], [m[2] for m in metrics], [m[3] for m in metrics]
total_pred, total_gt = sum([m[4] for m in metrics]), sum([m[5] for m in metrics])
current_result_row = [
config["name"],
total_gt, total_pred,
f"{np.mean(ious):.4f}", f"{np.mean(precs):.4f}", f"{np.mean(recs):.4f}", f"{np.mean(f1s):.4f}"
]
aggregate_table.append(current_result_row)
# Immediate output for this run
print(f"\n" + "="*80)
print(f" EXPERIMENT RESULTS: {config['name']}")
print(f"="*80)
print(tabulate([current_result_row], headers=headers, tablefmt="fancy_grid"))
print("\n")
# Visual Plot
pred_mask_sample, pred_count_sample = saved_masks[sample_filename]
plot_single_experiment_visual(
sample_filename=sample_filename,
combined_data_path=combined_data_path,
data_src_dir=dirs[config["data_src"]],
raw_data_dir=dirs["Raw"],
config_name=config["name"],
pred_mask=pred_mask_sample,
pred_count=pred_count_sample
)
# Final Summary Table
print("\n" + "=" * 115)
print(" " * 35 + f"FINAL 7-WAY EVALUATION REPORT (N={len(target_filenames)} images)")
print("=" * 115)
print(tabulate(aggregate_table, headers=headers, tablefmt="fancy_grid"))
if __name__ == "__main__":
main() |