def build_distance_layers(cyt_coord, nuc_coord, rna_coord, normalized=True):
"""Compute distance layers as input for the model.
Parameters
----------
cyt_coord : np.ndarray, np.int64
Array of cytoplasm boundaries coordinates with shape (nb_points, 2).
nuc_coord : np.ndarray, np.int64
Array of nucleus boundaries coordinates with shape (nb_points, 2).
rna_coord : np.ndarray, np.int64
Array of mRNAs coordinates with shape (nb_points, 2) or
(nb_points, 3).
normalized : bool
Normalized the layers between 0 and 1.
Returns
-------
distance_cyt : np.ndarray, np.float32
A 2-d tensor with shape (y, x) showing distance to the cytoplasm
border. Normalize between 0 and 1 if 'normalized' True.
distance_nuc : np.ndarray, np.float32
A 2-d tensor with shape (y, x) showing distance to the nucleus border.
Normalize between 0 and 1 if 'normalized' True.
"""
# check parameters
stack.check_array(cyt_coord,
ndim=2,
dtype=[np.int64])
if nuc_coord is not None:
stack.check_array(nuc_coord,
ndim=2,
dtype=[np.int64])
if rna_coord is not None:
stack.check_array(rna_coord,
ndim=2,
dtype=[np.int64])
stack.check_parameter(normalized=bool)
# build surface binary matrices from coordinates
cyt_surface, nuc_surface, rna_layer, _ = stack.from_coord_to_surface(
cyt_coord=cyt_coord,
nuc_coord=nuc_coord,
rna_coord=rna_coord)
# compute distances map for cytoplasm and nucleus
cyt_distance = ndi.distance_transform_edt(cyt_surface)
nuc_distance_ = ndi.distance_transform_edt(~nuc_surface)
nuc_distance = cyt_surface * nuc_distance_
if normalized:
# cast to np.float32 and normalize it between 0 and 1
cyt_distance = cyt_distance / cyt_distance.max()
nuc_distance = nuc_distance / nuc_distance.max()
# cast layer in float32
cyt_distance = stack.cast_img_float32(cyt_distance)
nuc_distance = stack.cast_img_float32(nuc_distance)
rna_layer = stack.cast_img_float32(rna_layer)
return cyt_distance, nuc_distance, rna_layer
def test_log_filter():
# float64
y_float64 = stack.cast_img_float64(y)
filtered_y_float64 = stack.log_filter(y_float64, 2)
expected_y_float64 = np.array(
[[0., 0., 0.02995949, 0.06212277, 0.07584532],
[0., 0., 0.02581818, 0.05134284, 0.06123539],
[0., 0., 0.01196859, 0.0253716, 0.02853162], [0., 0., 0., 0., 0.],
[0., 0., 0., 0., 0.]],
dtype=np.float64)
assert_allclose(filtered_y_float64, expected_y_float64, rtol=1e-6)
assert filtered_y_float64.dtype == np.float64
# float32
y_float32 = stack.cast_img_float32(y)
filtered_y = stack.log_filter(y_float32, 2)
expected_y = stack.cast_img_float32(expected_y_float64)
assert_allclose(filtered_y, expected_y, rtol=1e-6)
assert filtered_y.dtype == np.float32
# uint8
filtered_y = stack.log_filter(y, 2)
expected_y = stack.cast_img_uint8(expected_y_float64)
assert_array_equal(filtered_y, expected_y)
assert filtered_y.dtype == np.uint8
# uint16
y_uint16 = stack.cast_img_uint16(y)
filtered_y = stack.log_filter(y_uint16, 2)
expected_y = stack.cast_img_uint16(expected_y_float64)
assert_array_equal(filtered_y, expected_y)
assert filtered_y.dtype == np.uint16
def build_boundaries_layers(cyt_coord, nuc_coord, rna_coord):
"""
Parameters
----------
cyt_coord : np.ndarray, np.int64
Array of cytoplasm boundaries coordinates with shape (nb_points, 2).
nuc_coord : np.ndarray, np.int64
Array of nucleus boundaries coordinates with shape (nb_points, 2).
rna_coord : np.ndarray, np.int64
Array of mRNAs coordinates with shape (nb_points, 2) or
(nb_points, 3).
Returns
-------
cyt_boundaries : np.ndarray, np.float32
A 2-d binary tensor with shape (y, x) showing cytoplasm boundaries.
border.
nuc_boundaries : np.ndarray, np.float32
A 2-d binary tensor with shape (y, x) showing nucleus boundaries.
rna_layer : np.ndarray, np.float32
Binary image of mRNAs localizations with shape (y, x).
"""
# check parameters
stack.check_array(cyt_coord,
ndim=2,
dtype=[np.int64])
if nuc_coord is not None:
stack.check_array(nuc_coord,
ndim=2,
dtype=[np.int64])
if rna_coord is not None:
stack.check_array(rna_coord,
ndim=2,
dtype=[np.int64])
# build surface binary matrices from coordinates
cyt_surface, nuc_surface, rna_layer, _ = stack.from_coord_to_surface(
cyt_coord=cyt_coord,
nuc_coord=nuc_coord,
rna_coord=rna_coord)
# from surface binary matrices to boundaries binary matrices
cyt_boundaries = stack.from_surface_to_boundaries(cyt_surface)
nuc_boundaries = stack.from_surface_to_boundaries(nuc_surface)
# cast layer in float32
cyt_boundaries = stack.cast_img_float32(cyt_boundaries)
nuc_boundaries = stack.cast_img_float32(nuc_boundaries)
rna_layer = stack.cast_img_float32(rna_layer)
return cyt_boundaries, nuc_boundaries, rna_layer
def prepare_coordinate_data(cyt_coord, nuc_coord, rna_coord):
# get a binary representation of the coordinates
cyt, nuc = from_coord_to_matrix(cyt_coord, nuc_coord)
rna_coord[:, 1:3] += stack.get_offset_value()
# fill in masks
mask_cyt, mask_nuc = stack.get_surface_layers(cyt, nuc, cast_float=False)
# get mask cytoplasm outside nucleus
mask_cyt_out = mask_cyt.copy()
mask_cyt_out[mask_nuc] = False
# compute distance maps for the cytoplasm and the nucleus
distance_cyt, distance_nuc = stack.get_distance_layers(cyt,
nuc,
normalized=False)
# normalize distance maps between 0 and 1
distance_cyt_normalized = distance_cyt / distance_cyt.max()
distance_cyt_normalized = stack.cast_img_float32(distance_cyt_normalized)
distance_nuc_normalized = distance_nuc / distance_nuc.max()
distance_nuc_normalized = stack.cast_img_float32(distance_nuc_normalized)
# get rna outside nucleus
mask_rna_in = mask_nuc[rna_coord[:, 1], rna_coord[:, 2]]
rna_coord_out = rna_coord[~mask_rna_in]
# get centroids
centroid_cyt = get_centroid_surface(mask_cyt)
centroid_nuc = get_centroid_surface(mask_nuc)
centroid_rna = get_centroid_rna(rna_coord)
if len(rna_coord_out) == 0:
centroid_rna_out = centroid_cyt.copy()
else:
centroid_rna_out = get_centroid_rna(rna_coord_out)
# get centroid distance maps
distance_cyt_centroid = get_centroid_distance_map(centroid_cyt, mask_cyt)
distance_nuc_centroid = get_centroid_distance_map(centroid_nuc, mask_cyt)
distance_rna_out_centroid = get_centroid_distance_map(
centroid_rna_out, mask_cyt)
prepared_inputs = (mask_cyt, mask_nuc, mask_cyt_out, distance_cyt,
distance_nuc, distance_cyt_normalized,
distance_nuc_normalized, rna_coord_out, centroid_cyt,
centroid_nuc, centroid_rna, centroid_rna_out,
distance_cyt_centroid, distance_nuc_centroid,
distance_rna_out_centroid)
return prepared_inputs
def test_cast_float32(dtype):
# from integer to np.float32
if np.issubdtype(dtype, np.integer):
x = [[1, 2, 3], [4, 5, 6], [7, 8, 9]]
tensor = np.array(x).reshape((3, 3)).astype(dtype)
tensor[0, 0] = np.iinfo(dtype).min
tensor[2, 2] = np.iinfo(dtype).max
# from float to np.float32
else:
x = [[0.1, 0.2, 0.3], [0.4, 0.5, 0.6], [0.7, 0.8, 1.0]]
tensor = np.array(x).reshape((3, 3)).astype(dtype)
# cast in float32
tensor_float32 = stack.cast_img_float32(tensor)
assert tensor_float32.dtype == np.float32
# check value
if dtype in [np.uint8, np.uint16, np.uint32, np.uint64]:
assert tensor_float32.min() == 0
assert tensor_float32.max() == 1
elif dtype in [np.int8, np.int16, np.int32, np.int64]:
assert tensor_float32.min() == -1
assert tensor_float32.max() == 1
else:
assert_array_almost_equal(tensor_float32, tensor)
def test_cast_float32(dtype):
# from integer to np.float32
if np.issubdtype(dtype, np.integer):
x = [[1, 2, 3], [4, 5, 6], [7, 8, 9]]
tensor = np.array(x).reshape((3, 3)).astype(dtype)
tensor[2, 2] = np.iinfo(dtype).max
# from float to np.float32
else:
x = [[0.1, 0.2, 0.3], [0.4, 0.5, 0.6], [0.7, 0.8, 1.0]]
tensor = np.array(x).reshape((3, 3)).astype(dtype)
# cast in float32
if dtype == np.float64:
with pytest.warns(UserWarning):
tensor_float32 = stack.cast_img_float32(tensor)
else:
tensor_float32 = stack.cast_img_float32(tensor)
assert tensor_float32.dtype == np.float32
def test_gaussian_filter():
# float64
y_float64 = stack.cast_img_float64(y)
filtered_y_float64 = stack.gaussian_filter(y_float64, 2)
expected_y_float64 = np.array(
[[0.08928096, 0.1573019, 0.22897881, 0.28086597, 0.3001061],
[0.08668051, 0.14896399, 0.21282558, 0.25752308, 0.27253406],
[0.07634613, 0.12664142, 0.17574502, 0.20765944, 0.2155001],
[0.05890843, 0.09356377, 0.12493327, 0.1427122, 0.14374558],
[0.03878372, 0.05873308, 0.07492625, 0.08201409, 0.07939603]],
dtype=np.float64)
assert_allclose(filtered_y_float64, expected_y_float64, rtol=1e-6)
assert filtered_y_float64.dtype == np.float64
# float32
y_float32 = stack.cast_img_float32(y)
filtered_y = stack.gaussian_filter(y_float32, 2)
expected_y = stack.cast_img_float32(expected_y_float64)
assert_allclose(filtered_y, expected_y, rtol=1e-6)
assert filtered_y.dtype == np.float32
# uint8
with pytest.raises(ValueError):
stack.gaussian_filter(y, 2, allow_negative=True)
filtered_y = stack.gaussian_filter(y, 2)
expected_y = stack.cast_img_uint8(expected_y_float64)
assert_array_equal(filtered_y, expected_y)
assert filtered_y.dtype == np.uint8
# uint16
y_uint16 = stack.cast_img_uint16(y)
with pytest.raises(ValueError):
stack.gaussian_filter(y_uint16, 2, allow_negative=True)
filtered_y = stack.gaussian_filter(y_uint16, 2)
expected_y = stack.cast_img_uint16(expected_y_float64)
assert_array_equal(filtered_y, expected_y)
assert filtered_y.dtype == np.uint16
def test_background_removal_gaussian():
# float64
y_float64 = stack.cast_img_float64(y)
filtered_y_float64 = stack.remove_background_gaussian(y_float64, 2)
expected_y_float64 = np.array(
[[0., 0., 0.01415845, 0.36227129, 0.],
[0., 0., 0.25776265, 0.66404555, 0.43726986],
[0., 0., 0.11052949, 0.59626213, 0.], [0., 0.42016172, 0., 0., 0.],
[0., 0., 0., 0., 0.]],
dtype=np.float64)
assert_allclose(filtered_y_float64, expected_y_float64, rtol=1e-6)
assert filtered_y_float64.dtype == np.float64
# float32
y_float32 = stack.cast_img_float32(y)
filtered_y = stack.remove_background_gaussian(y_float32, 2)
expected_y = stack.cast_img_float32(expected_y_float64)
assert_allclose(filtered_y, expected_y, rtol=1e-6)
assert filtered_y.dtype == np.float32
# uint8
with pytest.raises(ValueError):
stack.gaussian_filter(y, 2, allow_negative=True)
filtered_y = stack.remove_background_gaussian(y, 2)
expected_y = stack.cast_img_uint8(expected_y_float64)
assert_array_equal(filtered_y, expected_y)
assert filtered_y.dtype == np.uint8
# uint16
y_uint16 = stack.cast_img_uint16(y)
with pytest.raises(ValueError):
stack.gaussian_filter(y_uint16, 2, allow_negative=True)
filtered_y = stack.remove_background_gaussian(y_uint16, 2)
expected_y = stack.cast_img_uint16(expected_y_float64)
assert_array_equal(filtered_y, expected_y)
assert filtered_y.dtype == np.uint16
def build_cyt_relief(image_projected, nuc_labelled, mask_cyt, alpha=0.8):
"""Compute a 2-d representation of the cytoplasm to be used by watershed
algorithm.
Cells are represented as watershed, with a low values to the center and
maximum values at their borders.
The equation used is:
relief = alpha * relief_pixel + (1 - alpha) * relief_distance
- 'relief_pixel' exploit the differences in pixel intensity values.
- 'relief_distance' use the distance from the nuclei.
Parameters
----------
image_projected : np.ndarray, np.uint
Projected image of the cytoplasm with shape (y, x).
nuc_labelled : np.ndarray,
Result of the nuclei segmentation with shape (y, x).
mask_cyt : np.ndarray, bool
Binary mask of the cytoplasm with shape (y, x).
alpha : float or int
Weight of the pixel intensity values to compute the relief. A value of
0 and 1 respectively return 'relief_distance' and 'relief_pixel'.
Returns
-------
relief : np.ndarray, np.uint
Relief image of the cytoplasm with shape (y, x).
"""
# check parameters
stack.check_array(image_projected,
ndim=2,
dtype=[np.uint8, np.uint16])
stack.check_array(nuc_labelled,
ndim=2,
dtype=[np.uint8, np.uint16, np.int64, bool])
stack.check_array(mask_cyt,
ndim=2,
dtype=[bool])
stack.check_parameter(alpha=(float, int))
# use pixel intensity of the cytoplasm channel to compute the seed.
if alpha == 1:
relief = image_projected.copy()
max_intensity = np.iinfo(image_projected.dtype).max
relief = max_intensity - relief
relief[nuc_labelled > 0] = 0
relief[mask_cyt == 0] = max_intensity
relief = stack.rescale(relief)
# use distance from the nuclei
elif alpha == 0:
binary_mask_nuc = nuc_labelled > 0
relief = ndi.distance_transform_edt(~binary_mask_nuc)
relief[mask_cyt == 0] = relief.max()
relief = np.true_divide(relief, relief.max(), dtype=np.float32)
if image_projected.dtype == np.uint8:
relief = stack.cast_img_uint8(relief)
else:
relief = stack.cast_img_uint16(relief)
# use both previous methods
elif 0 < alpha < 1:
relief_pixel = image_projected.copy()
max_intensity = np.iinfo(image_projected.dtype).max
relief_pixel = max_intensity - relief_pixel
relief_pixel[nuc_labelled > 0] = 0
relief_pixel[mask_cyt == 0] = max_intensity
relief_pixel = stack.rescale(relief_pixel)
relief_pixel = stack.cast_img_float32(relief_pixel)
binary_mask_nuc = nuc_labelled > 0
relief_distance = ndi.distance_transform_edt(~binary_mask_nuc)
relief_distance[mask_cyt == 0] = relief_distance.max()
relief_distance = np.true_divide(relief_distance,
relief_distance.max(),
dtype=np.float32)
relief = alpha * relief_pixel + (1 - alpha) * relief_distance
if image_projected.dtype == np.uint8:
relief = stack.cast_img_uint8(relief)
else:
relief = stack.cast_img_uint16(relief)
else:
raise ValueError("Parameter 'alpha' is wrong. Must be comprised "
"between 0 and 1. Currently 'alpha' is {0}"
.format(alpha))
return relief
def prepare_coordinate_data(cyt_coord, nuc_coord, rna_coord):
"""
Parameters
----------
cyt_coord
nuc_coord
rna_coord
Returns
-------
"""
# convert coordinates in binary mask surfaces
mask_cyt, mask_nuc, _, rna_coord = stack.from_coord_to_surface(
cyt_coord=cyt_coord,
nuc_coord=nuc_coord,
rna_coord=rna_coord,
external_coord=True)
# get mask cytoplasm outside nucleus
mask_cyt_out = mask_cyt.copy()
mask_cyt_out[mask_nuc] = False
# compute distance maps for the cytoplasm and the nucleus
distance_cyt = ndi.distance_transform_edt(mask_cyt)
distance_nuc_ = ndi.distance_transform_edt(~mask_nuc)
distance_nuc = mask_cyt * distance_nuc_
# cast distance map in float32
distance_cyt = distance_cyt.astype(np.float32)
distance_nuc = distance_nuc.astype(np.float32)
# normalize distance maps between 0 and 1
distance_cyt_normalized = distance_cyt / distance_cyt.max()
distance_cyt_normalized = stack.cast_img_float32(distance_cyt_normalized)
distance_nuc_normalized = distance_nuc / distance_nuc.max()
distance_nuc_normalized = stack.cast_img_float32(distance_nuc_normalized)
# get rna outside nucleus
mask_rna_in = mask_nuc[rna_coord[:, 1], rna_coord[:, 2]]
rna_coord_out = rna_coord[~mask_rna_in]
# get centroids
centroid_cyt = _get_centroid_surface(mask_cyt)
centroid_nuc = _get_centroid_surface(mask_nuc)
if len(rna_coord) == 0:
centroid_rna = centroid_cyt.copy()
else:
centroid_rna = _get_centroid_rna(rna_coord)
if len(rna_coord_out) == 0:
centroid_rna_out = centroid_cyt.copy()
else:
centroid_rna_out = _get_centroid_rna(rna_coord_out)
# get centroid distance maps
distance_cyt_centroid = _get_centroid_distance_map(centroid_cyt, mask_cyt)
distance_nuc_centroid = _get_centroid_distance_map(centroid_nuc, mask_cyt)
distance_rna_out_centroid = _get_centroid_distance_map(
centroid_rna_out, mask_cyt)
prepared_inputs = (mask_cyt, mask_nuc, mask_cyt_out, distance_cyt,
distance_nuc, distance_cyt_normalized,
distance_nuc_normalized, rna_coord_out, centroid_cyt,
centroid_nuc, centroid_rna, centroid_rna_out,
distance_cyt_centroid, distance_nuc_centroid,
distance_rna_out_centroid)
return prepared_inputs
