def write_thread(out_file_path: Path,
data: np.ndarray,
metadata: OmeXml,
chan_name: str):
""" Thread for saving images
This function is intended to be run inside a threadpool to save an image.
Args:
out_file_path (Path): Path to an output file
data (np.ndarray): FOV to save
metadata (OmeXml): Metadata for the image
chan_name (str): Name of the channel
"""
ProcessManager.log(f'Writing: {out_file_path.name}')
with BioWriter(out_file_path,metadata=metadata) as bw:
bw.X = data.shape[1]
bw.Y = data.shape[0]
bw.Z = 1
bw.C = 1
bw.cnames = [chan_name]
bw[:] = data
def assemble_image(vector_path: pathlib.Path, out_path: pathlib.Path,
depth: int) -> None:
"""Assemble a 2d or 3d image
This method assembles one image from one stitching vector. It can
assemble both 2d and z-stacked 3d images It is intended to run as
a process to parallelize stitching of multiple images.
The basic approach to stitching is:
1. Parse the stitching vector and abstract the image dimensions
2. Generate a thread for each subsection (supertile) of an image.
Args:
vector_path: Path to the stitching vector
out_path: Path to the output directory
depth: depth of the input images
"""
# Grab a free process
with ProcessManager.process():
# Parse the stitching vector
parsed_vector = _parse_stitch(vector_path, timesliceNaming)
# Initialize the output image
with BioReader(parsed_vector['filePos'][0]['file']) as br:
bw = BioWriter(out_path.joinpath(parsed_vector['name']),
metadata=br.metadata,
max_workers=ProcessManager._active_threads)
bw.x = parsed_vector['width']
bw.y = parsed_vector['height']
bw.z = depth
# Assemble the images
ProcessManager.log(f'Begin assembly')
for z in range(depth):
ProcessManager.log(f'Assembling Z position : {z}')
for x in range(0, parsed_vector['width'], chunk_size):
X_range = min(x + chunk_size, parsed_vector['width']
) # max x-pixel index in the assembled image
for y in range(0, parsed_vector['height'], chunk_size):
Y_range = min(y + chunk_size, parsed_vector['height']
) # max y-pixel index in the assembled image
ProcessManager.submit_thread(make_tile, x, X_range, y,
Y_range, z, parsed_vector, bw)
ProcessManager.join_threads()
bw.close()
def label_cython(input_path: Path, output_path: Path, connectivity: int):
""" Label the input image and writes labels back out.
Args:
input_path: Path to input image.
output_path: Path for output image.
connectivity: Connectivity kind.
"""
with ProcessManager.thread() as active_threads:
with BioReader(
input_path,
max_workers=active_threads.count,
) as reader:
with BioWriter(
output_path,
max_workers=active_threads.count,
metadata=reader.metadata,
) as writer:
# Load an image and convert to binary
image = numpy.squeeze(reader[..., 0, 0])
if not numpy.any(image):
writer.dtype = numpy.uint8
writer[:] = numpy.zeros_like(image, dtype=numpy.uint8)
return
image = (image > 0)
if connectivity > image.ndim:
ProcessManager.log(
f'{input_path.name}: Connectivity is not less than or equal to the number of image dimensions, '
f'skipping this image. connectivity={connectivity}, ndim={image.ndim}'
)
return
# Run the labeling algorithm
labels = ftl.label_nd(image, connectivity)
# Save the image
writer.dtype = labels.dtype
writer[:] = labels
return True
def label_thread(input_path, output_path, connectivity):
with ProcessManager.thread() as active_threads:
with bfio.BioReader(input_path,
max_workers=active_threads.count) as br:
with bfio.BioWriter(output_path,
max_workers=active_threads.count,
metadata=br.metadata) as bw:
# Load an image and convert to binary
image = (br[..., 0, 0] > 0).squeeze()
if connectivity > image.ndim:
ProcessManager.log(
"{}: Connectivity is not less than or equal to the number of image dimensions, skipping this image. connectivity={}, ndim={}"
.format(input_path.name, connectivity, image.ndim))
return
# Run the labeling algorithm
labels = ftl.label_nd(image.squeeze(), connectivity)
# Save the image
bw.dtype = labels.dtype
bw[:] = labels
def _parse_stitch(stitchPath: pathlib.Path,
timepointName: bool = False) -> dict:
""" Load and parse image stitching vectors
This function parses the data from a stitching vector, then extracts the
relevant image sizes for each image in the stitching vector to obtain a
stitched image size. This function also infers an output file name.
Args:
stitchPath: A path to stitching vectors
timepointName: Use the vector timeslice as the image name
Returns:
Dictionary with keys (width, height, name, filePos)
"""
# Initialize the output
out_dict = { 'width': int(0),
'height': int(0),
'name': '',
'filePos': []}
# Try to parse the stitching vector using the infered file pattern
if fp.pattern != '.*':
vp = filepattern.VectorPattern(stitchPath,fp.pattern)
unique_vals = {k.upper():v for k,v in vp.uniques.items() if len(v)==1}
files = fp.get_matching(**unique_vals)
else:
# Try to infer a pattern from the stitching vector
try:
vector_files = filepattern.VectorPattern(stitchPath,'.*')
pattern = filepattern.infer_pattern([v[0]['file'] for v in vector_files()])
vp = filepattern.VectorPattern(stitchPath,pattern)
# Fall back to universal filepattern
except ValueError:
vp = filepattern.VectorPattern(stitchPath,'.*')
files = fp.files
file_names = [f['file'].name for f in files]
for file in vp():
if file[0]['file'] not in file_names:
continue
stitch_groups = {k:get_number(v) for k,v in file[0].items()}
stitch_groups['file'] = files[0]['file'].with_name(stitch_groups['file'])
# Get the image size
stitch_groups['width'], stitch_groups['height'] = BioReader.image_size(stitch_groups['file'])
# Set the stitching vector values in the file dictionary
out_dict['filePos'].append(stitch_groups)
# Calculate the output image dimensions
out_dict['width'] = max([f['width'] + f['posX'] for f in out_dict['filePos']])
out_dict['height'] = max([f['height'] + f['posY'] for f in out_dict['filePos']])
# Generate the output file name
if timepointName:
global_regex = ".*global-positions-([0-9]+).txt"
name = re.match(global_regex,pathlib.Path(stitchPath).name).groups()[0]
name += '.ome.tif'
out_dict['name'] = name
ProcessManager.job_name(out_dict['name'])
ProcessManager.log(f'Setting output name to timepoint slice number.')
else:
# Try to infer a good filename
try:
out_dict['name'] = vp.output_name()
ProcessManager.job_name(out_dict['name'])
ProcessManager.log(f'Inferred output file name from vector.')
# A file name couldn't be inferred, default to the first image name
except:
ProcessManager.job_name(out_dict['name'])
ProcessManager.log(f'Could not infer output file name from vector, using first file name in the stitching vector as an output file name.')
for file in vp():
out_dict['name'] = file[0]['file']
break
return out_dict
def basic(files: typing.List[Path],
out_dir: Path,
metadata_dir: typing.Optional[Path] = None,
darkfield: bool = False,
photobleach: bool = False):
# Try to infer a filename
try:
pattern = infer_pattern([f['file'].name for f in files])
fp = FilePattern(files[0]['file'].parent,pattern)
base_output = fp.output_name()
# Fallback to the first filename
except:
base_output = files[0]['file'].name
extension = ''.join(files[0]['file'].suffixes)
with ProcessManager.process(base_output):
# Load files and sort
ProcessManager.log('Loading and sorting images...')
img_stk,X,Y = _get_resized_image_stack(files)
img_stk_sort = np.sort(img_stk)
# Initialize options
new_options = _initialize_options(img_stk_sort,darkfield,OPTIONS)
# Initialize flatfield/darkfield matrices
ProcessManager.log('Beginning flatfield estimation')
flatfield_old = np.ones((new_options['size'],new_options['size']),dtype=np.float64)
darkfield_old = np.random.normal(size=(new_options['size'],new_options['size'])).astype(np.float64)
# Optimize until the change in values is below tolerance or a maximum number of iterations is reached
for w in range(new_options['max_reweight_iterations']):
# Optimize using inexact augmented Legrangian multiplier method using L1 loss
A, E1, A_offset = _inexact_alm_l1(copy.deepcopy(img_stk_sort),new_options)
# Calculate the flatfield/darkfield images and update training weights
flatfield, darkfield, new_options = _get_flatfield_and_reweight(A,E1,A_offset,new_options)
# Calculate the change in flatfield and darkfield images between iterations
mad_flat = np.sum(np.abs(flatfield-flatfield_old))/np.sum(np.abs(flatfield_old))
temp_diff = np.sum(np.abs(darkfield - darkfield_old))
if temp_diff < 10**-7:
mad_dark =0
else:
mad_dark = temp_diff/np.max(np.sum(np.abs(darkfield_old)),initial=10**-6)
flatfield_old = flatfield
darkfield_old = darkfield
# Stop optimizing if the change in flatfield/darkfield is below threshold
ProcessManager.log('Iteration {} loss: {}'.format(w+1,mad_flat))
if np.max(mad_flat,initial=mad_dark) < new_options['reweight_tol']:
break
# Calculate photobleaching effects if specified
if photobleach:
pb = _get_photobleach(copy.deepcopy(img_stk),flatfield,darkfield)
# Resize images back to original image size
ProcessManager.log('Saving outputs...')
flatfield = cv2.resize(flatfield,(Y,X),interpolation=cv2.INTER_CUBIC).astype(np.float32)
if new_options['darkfield']:
darkfield = cv2.resize(darkfield,(Y,X),interpolation=cv2.INTER_CUBIC).astype(np.float32)
# Export the flatfield image as a tiled tiff
flatfield_out = base_output.replace(extension,'_flatfield' + extension)
with BioReader(files[0]['file'],max_workers=2) as br:
metadata = br.metadata
with BioWriter(out_dir.joinpath(flatfield_out),metadata=metadata,max_workers=2) as bw:
bw.dtype = np.float32
bw.x = X
bw.y = Y
bw[:] = np.reshape(flatfield,(Y,X,1,1,1))
# Export the darkfield image as a tiled tiff
if new_options['darkfield']:
darkfield_out = base_output.replace(extension,'_darkfield' + extension)
with BioWriter(out_dir.joinpath(darkfield_out),metadata=metadata,max_workers=2) as bw:
bw.dtype = np.float32
bw.x = X
bw.y = Y
bw[:] = np.reshape(darkfield,(Y,X,1,1,1))
# Export the photobleaching components as csv
if photobleach:
offsets_out = base_output.replace(extension,'_offsets.csv')
with open(metadata_dir.joinpath(offsets_out),'w') as fw:
fw.write('file,offset\n')
for f,o in zip(files,pb[0,:].tolist()):
fw.write("{},{}\n".format(f,o))
def _merge_layers(input_files,output_path):
with ProcessManager.process(output_path.name):
# Get the number of layers to stack
z_size = 0
for f in input_files:
with BioReader(f['file']) as br:
z_size += br.z
# Get some basic info about the files to stack
with BioReader(input_files[0]['file']) as br:
# Get the physical z-distance if available, set to physical x if not
ps_z = br.ps_z
# If the z-distances are undefined, average the x and y together
if None in ps_z:
# Get the size and units for x and y
x_val,x_units = br.ps_x
y_val,y_units = br.ps_y
# Convert x and y values to the same units and average
z_val = (x_val*UNITS[x_units] + y_val*UNITS[y_units])/2
# Set z units to the smaller of the units between x and y
z_units = x_units if UNITS[x_units] < UNITS[y_units] else y_units
# Convert z to the proper unit scale
z_val /= UNITS[z_units]
ps_z = (z_val,z_units)
ProcessManager.log('Could not find physical z-size. Using the average of x & y {}.'.format(ps_z))
# Hold a reference to the metadata once the file gets closed
metadata = br.metadata
# Create the output file within a context manager
with BioWriter(output_path,metadata=metadata,max_workers=ProcessManager._active_threads) as bw:
# Adjust the dimensions before writing
bw.z = z_size
bw.ps_z = ps_z
# ZIndex tracking for the output file
zi = 0
# Start stacking
for file in input_files:
# Open an image
with BioReader(file['file'],max_workers=ProcessManager._active_threads) as br:
# Open z-layers one at a time
for z in range(br.z):
# Use tiled reading in x&y to conserve memory
# At most, [chunk_size, chunk_size] pixels are loaded
for xs in range(0,br.x,chunk_size):
xe = min([br.x,xs + chunk_size])
for ys in range(0,br.y,chunk_size):
ye = min([br.y,ys + chunk_size])
bw[ys:ye,xs:xe,zi:zi+1,...] = br[ys:ye,xs:xe,z:z+1,...]
zi += 1
# update the BioWriter in case the ProcessManager found more threads
bw.max_workers = ProcessManager._active_threads
