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16 changes: 9 additions & 7 deletions starfish/core/spots/FindSpots/blob.py
Original file line number Diff line number Diff line change
Expand Up @@ -251,13 +251,15 @@ def run(
merged_z_tables[(r, ch)] = pd.concat(
[merged_z_tables[(r, ch)], spot_attributes_list[i][0].spot_attrs.data])
new = []
r_chs = sorted([*merged_z_tables])
selectors = list(image_stack._iter_axes({Axes.ROUND, Axes.CH}))
for i, (r, ch) in enumerate(r_chs):
merged_z_tables[(r, ch)]['spot_id'] = range(len(merged_z_tables[(r, ch)]))
spot_attrs = SpotAttributes(merged_z_tables[(r, ch)].reset_index(drop=True))
new.append((PerImageSliceSpotResults(spot_attrs=spot_attrs, extras=None),
selectors[i]))
# Iterate through the merged tables in the order expected by _iter_axes
for selector in image_stack._iter_axes({Axes.ROUND, Axes.CH}):
r = selector[Axes.ROUND]
ch = selector[Axes.CH]
if (r, ch) in merged_z_tables:
merged_z_tables[(r, ch)]['spot_id'] = range(len(merged_z_tables[(r, ch)]))
spot_attrs = SpotAttributes(merged_z_tables[(r, ch)].reset_index(drop=True))
new.append((PerImageSliceSpotResults(spot_attrs=spot_attrs, extras=None),
selector))

spot_attributes_list = new

Expand Down
90 changes: 90 additions & 0 deletions starfish/core/spots/FindSpots/test/test_spot_detection.py
Original file line number Diff line number Diff line change
Expand Up @@ -263,3 +263,93 @@ def test_blob_detector_2d_with_reference_image():
# Check that radius is reasonable
assert np.all(radius_values < 100), f"Radius values too large: {radius_values}"
assert np.all(radius_values > 0), f"Radius values should be positive: {radius_values}"


def test_blob_detector_round_channel_assignment():
"""Test that BlobDetector with is_volume=False assigns spots to correct (round, channel) pairs.

This is a regression test for a bug where spots were assigned to incorrect round/channel
combinations due to a mismatch between the sorted (r, ch) tuples and the iteration order
from _iter_axes({Axes.ROUND, Axes.CH}).
"""
# Create a test ImageStack with 2 rounds, 3 channels, 1 z-plane
# Each (round, channel) pair gets a spot at a unique location
data = np.zeros((2, 3, 1, 100, 100), dtype=np.float32)

# Add spots at different y-coordinates for each (round, channel)
# Round 0, Channel 0: y=10
data[0, 0, 0, 8:13, 8:13] = 1.0
# Round 0, Channel 1: y=20
data[0, 1, 0, 18:23, 18:23] = 1.0
# Round 0, Channel 2: y=30
data[0, 2, 0, 28:33, 28:33] = 1.0
# Round 1, Channel 0: y=40
data[1, 0, 0, 38:43, 38:43] = 1.0
# Round 1, Channel 1: y=50
data[1, 1, 0, 48:53, 48:53] = 1.0
# Round 1, Channel 2: y=60
data[1, 2, 0, 58:63, 58:63] = 1.0

image_stack = ImageStack.from_numpy(data)

# Test with is_volume=False
detector_2d = BlobDetector(
min_sigma=1,
max_sigma=3,
num_sigma=5,
threshold=0.01,
is_volume=False,
measurement_type='mean'
)

spots_2d = detector_2d.run(image_stack=image_stack)

# Test with is_volume=True for comparison
detector_3d = BlobDetector(
min_sigma=1,
max_sigma=3,
num_sigma=5,
threshold=0.01,
is_volume=True,
measurement_type='mean'
)

spots_3d = detector_3d.run(image_stack=image_stack)

# Define expected y-coordinates for each (round, channel) pair
expected_y = {
(0, 0): 10,
(0, 1): 20,
(0, 2): 30,
(1, 0): 40,
(1, 1): 50,
(1, 2): 60,
}

# Check that both is_volume=False and is_volume=True produce the same results
for r in range(2):
for ch in range(3):
# Get spots for this (round, channel)
spots_2d_data = spots_2d[{Axes.ROUND: r, Axes.CH: ch}].spot_attrs.data
spots_3d_data = spots_3d[{Axes.ROUND: r, Axes.CH: ch}].spot_attrs.data

# Both should have found the spot
assert len(spots_2d_data) > 0, \
f"No spots found for Round={r}, CH={ch} with is_volume=False"
assert len(spots_3d_data) > 0, \
f"No spots found for Round={r}, CH={ch} with is_volume=True"

# Check that the y-coordinate matches the expected value
y_2d = spots_2d_data['y'].values[0]
y_3d = spots_3d_data['y'].values[0]
expected = expected_y[(r, ch)]

assert np.abs(y_2d - expected) < 5, \
f"is_volume=False: Round={r}, CH={ch} has y={y_2d:.0f}, expected ~{expected}"
assert np.abs(y_3d - expected) < 5, \
f"is_volume=True: Round={r}, CH={ch} has y={y_3d:.0f}, expected ~{expected}"

# Both should produce the same y-coordinate
assert np.abs(y_2d - y_3d) < 2, \
f"Round={r}, CH={ch}: is_volume=False has y={y_2d:.0f} " \
f"but is_volume=True has y={y_3d:.0f}"