def add_image(nwbfile, image_data, image_name, module_name, module_description, image_api=None):
description = '{} image at pixels/cm resolution'.format(image_name)
if image_api is None:
image_api = ImageApi
if isinstance(image_data, sitk.Image):
data, spacing, unit = ImageApi.deserialize(image_data)
elif isinstance(image_data, Image):
data = image_data.data
spacing = image_data.spacing
unit = image_data.unit
else:
raise ValueError("Not a supported image_data type: {}".format(type(image_data)))
assert spacing[0] == spacing[1] and len(spacing) == 2 and unit == 'mm'
if module_name not in nwbfile.modules:
ophys_mod = ProcessingModule(module_name, module_description)
nwbfile.add_processing_module(ophys_mod)
else:
ophys_mod = nwbfile.modules[module_name]
image = GrayscaleImage(image_name, data, resolution=spacing[0] / 10, description=description)
if 'images' not in ophys_mod.containers:
images = Images(name='images')
ophys_mod.add_data_interface(images)
else:
images = ophys_mod['images']
images.add_image(image)
return nwbfile
def add_images_to_subject(subject, subject_image_list):
images = Images(name='images', description="images of subject's brain")
for image_path in subject_image_list:
image_name = os.path.split(image_path)[1]
image_name = os.path.splitext(image_name)[0]
image_data = imread(image_path)
kwargs = {'data': image_data, 'name': image_name}
if len(image_data.shape) == 2:
image = GrayscaleImage(**kwargs)
elif image_data.shape[2] == 3:
image = RGBImage(**kwargs)
elif image_data.shape[2] == 4:
image = RGBAImage(**kwargs)
images.add_image(image)
subject.images = images
return subject
def add_image_to_nwb(nwbfile: NWBFile, image_data: Image, image_name: str):
"""
Adds image given by image_data with name image_name to nwbfile
Parameters
----------
nwbfile
nwbfile to add image to
image_data
The image data
image_name
Image name
Returns
-------
None
"""
module_name = 'ophys'
description = '{} image at pixels/cm resolution'.format(image_name)
data, spacing, unit = image_data
assert spacing[0] == spacing[1] and len(spacing) == 2 and unit == 'mm'
if module_name not in nwbfile.processing:
ophys_mod = ProcessingModule(module_name, 'Ophys processing module')
nwbfile.add_processing_module(ophys_mod)
else:
ophys_mod = nwbfile.processing[module_name]
image = GrayscaleImage(image_name,
data,
resolution=spacing[0] / 10,
description=description)
if 'images' not in ophys_mod.containers:
images = Images(name='images')
ophys_mod.add_data_interface(images)
else:
images = ophys_mod['images']
images.add_image(image)
def test_grayscale_image(self):
GrayscaleImage(name='test_grayscale_image', data=np.ones((2, 2)))
def write_segmentation(
segext_obj: SegmentationExtractor,
save_path: PathType = None,
plane_num=0,
metadata: dict = None,
overwrite: bool = True,
buffer_size: int = 10,
nwbfile=None,
):
assert (
save_path is None or nwbfile is None
), "Either pass a save_path location, or nwbfile object, but not both!"
# parse metadata correctly:
if isinstance(segext_obj, MultiSegmentationExtractor):
segext_objs = segext_obj.segmentations
if metadata is not None:
assert isinstance(metadata, list), (
"For MultiSegmentationExtractor enter 'metadata' as a list of "
"SegmentationExtractor metadata")
assert len(metadata) == len(segext_objs), (
"The 'metadata' argument should be a list with the same "
"number of elements as the segmentations in the "
"MultiSegmentationExtractor")
else:
segext_objs = [segext_obj]
if metadata is not None and not isinstance(metadata, list):
metadata = [metadata]
metadata_base_list = [
NwbSegmentationExtractor.get_nwb_metadata(sgobj)
for sgobj in segext_objs
]
print(f"writing nwb for {segext_obj.extractor_name}\n")
# updating base metadata with new:
for num, data in enumerate(metadata_base_list):
metadata_input = metadata[num] if metadata else {}
metadata_base_list[num] = dict_recursive_update(
metadata_base_list[num], metadata_input)
metadata_base_common = metadata_base_list[0]
# build/retrieve nwbfile:
if nwbfile is not None:
assert isinstance(
nwbfile, NWBFile), "'nwbfile' should be of type pynwb.NWBFile"
write = False
else:
write = True
save_path = Path(save_path)
assert save_path.suffix == ".nwb"
if save_path.is_file() and not overwrite:
nwbfile_exist = True
file_mode = "r+"
else:
if save_path.is_file():
os.remove(save_path)
if not save_path.parent.is_dir():
save_path.parent.mkdir(parents=True)
nwbfile_exist = False
file_mode = "w"
io = NWBHDF5IO(str(save_path), file_mode)
if nwbfile_exist:
nwbfile = io.read()
else:
nwbfile = NWBFile(**metadata_base_common["NWBFile"])
# Subject:
if metadata_base_common.get("Subject") and nwbfile.subject is None:
nwbfile.subject = Subject(**metadata_base_common["Subject"])
# Processing Module:
if "ophys" not in nwbfile.processing:
ophys = nwbfile.create_processing_module(
"ophys", "contains optical physiology processed data")
else:
ophys = nwbfile.get_processing_module("ophys")
for plane_no_loop, (segext_obj, metadata) in enumerate(
zip(segext_objs, metadata_base_list)):
# Device:
if metadata["Ophys"]["Device"][0]["name"] not in nwbfile.devices:
nwbfile.create_device(**metadata["Ophys"]["Device"][0])
# ImageSegmentation:
image_segmentation_name = (
"ImageSegmentation" if plane_no_loop == 0 else
f"ImageSegmentation_Plane{plane_no_loop}")
if image_segmentation_name not in ophys.data_interfaces:
image_segmentation = ImageSegmentation(
name=image_segmentation_name)
ophys.add(image_segmentation)
else:
image_segmentation = ophys.data_interfaces.get(
image_segmentation_name)
# OpticalChannel:
optical_channels = [
OpticalChannel(**i) for i in metadata["Ophys"]["ImagingPlane"]
[0]["optical_channel"]
]
# ImagingPlane:
image_plane_name = ("ImagingPlane" if plane_no_loop == 0 else
f"ImagePlane_{plane_no_loop}")
if image_plane_name not in nwbfile.imaging_planes.keys():
input_kwargs = dict(
name=image_plane_name,
device=nwbfile.get_device(
metadata_base_common["Ophys"]["Device"][0]["name"]),
)
metadata["Ophys"]["ImagingPlane"][0][
"optical_channel"] = optical_channels
input_kwargs.update(**metadata["Ophys"]["ImagingPlane"][0])
if "imaging_rate" in input_kwargs:
input_kwargs["imaging_rate"] = float(
input_kwargs["imaging_rate"])
imaging_plane = nwbfile.create_imaging_plane(**input_kwargs)
else:
imaging_plane = nwbfile.imaging_planes[image_plane_name]
# PlaneSegmentation:
input_kwargs = dict(
description="output from segmenting imaging plane",
imaging_plane=imaging_plane,
)
ps_metadata = metadata["Ophys"]["ImageSegmentation"][
"plane_segmentations"][0]
if ps_metadata[
"name"] not in image_segmentation.plane_segmentations:
ps_exist = False
else:
ps = image_segmentation.get_plane_segmentation(
ps_metadata["name"])
ps_exist = True
roi_ids = segext_obj.get_roi_ids()
accepted_list = segext_obj.get_accepted_list()
accepted_list = [] if accepted_list is None else accepted_list
rejected_list = segext_obj.get_rejected_list()
rejected_list = [] if rejected_list is None else rejected_list
accepted_ids = [1 if k in accepted_list else 0 for k in roi_ids]
rejected_ids = [1 if k in rejected_list else 0 for k in roi_ids]
roi_locations = np.array(segext_obj.get_roi_locations()).T
def image_mask_iterator():
for id in segext_obj.get_roi_ids():
img_msks = segext_obj.get_roi_image_masks(
roi_ids=[id]).T.squeeze()
yield img_msks
if not ps_exist:
input_kwargs.update(
**ps_metadata,
columns=[
VectorData(
data=H5DataIO(
DataChunkIterator(image_mask_iterator(),
buffer_size=buffer_size),
compression=True,
compression_opts=9,
),
name="image_mask",
description="image masks",
),
VectorData(
data=roi_locations,
name="RoiCentroid",
description=
"x,y location of centroid of the roi in image_mask",
),
VectorData(
data=accepted_ids,
name="Accepted",
description=
"1 if ROi was accepted or 0 if rejected as a cell during segmentation operation",
),
VectorData(
data=rejected_ids,
name="Rejected",
description=
"1 if ROi was rejected or 0 if accepted as a cell during segmentation operation",
),
],
id=roi_ids,
)
ps = image_segmentation.create_plane_segmentation(
**input_kwargs)
# Fluorescence Traces:
if "Flourescence" not in ophys.data_interfaces:
fluorescence = Fluorescence()
ophys.add(fluorescence)
else:
fluorescence = ophys.data_interfaces["Fluorescence"]
roi_response_dict = segext_obj.get_traces_dict()
roi_table_region = ps.create_roi_table_region(
description=f"region for Imaging plane{plane_no_loop}",
region=list(range(segext_obj.get_num_rois())),
)
rate = (np.float("NaN")
if segext_obj.get_sampling_frequency() is None else
segext_obj.get_sampling_frequency())
for i, j in roi_response_dict.items():
data = getattr(segext_obj, f"_roi_response_{i}")
if data is not None:
data = np.asarray(data)
trace_name = "RoiResponseSeries" if i == "raw" else i.capitalize(
)
trace_name = (trace_name if plane_no_loop == 0 else
trace_name + f"_Plane{plane_no_loop}")
input_kwargs = dict(
name=trace_name,
data=data.T,
rois=roi_table_region,
rate=rate,
unit="n.a.",
)
if trace_name not in fluorescence.roi_response_series:
fluorescence.create_roi_response_series(**input_kwargs)
# create Two Photon Series:
if "TwoPhotonSeries" not in nwbfile.acquisition:
warn(
"could not find TwoPhotonSeries, using ImagingExtractor to create an nwbfile"
)
# adding images:
images_dict = segext_obj.get_images_dict()
if any([image is not None for image in images_dict.values()]):
images_name = ("SegmentationImages" if plane_no_loop == 0 else
f"SegmentationImages_Plane{plane_no_loop}")
if images_name not in ophys.data_interfaces:
images = Images(images_name)
for img_name, img_no in images_dict.items():
if img_no is not None:
images.add_image(
GrayscaleImage(name=img_name, data=img_no.T))
ophys.add(images)
# saving NWB file:
if write:
io.write(nwbfile)
io.close()
# test read
with NWBHDF5IO(str(save_path), "r") as io:
io.read()
def save_nwb(ops1):
if NWB and not ops1[0]['mesoscan']:
if len(ops1) > 1:
multiplane = True
else:
multiplane = False
ops = ops1[0]
### INITIALIZE NWB FILE
nwbfile = NWBFile(
session_description='suite2p_proc',
identifier=ops['data_path'][0],
session_start_time=(ops['date_proc'] if 'date_proc' in ops else
datetime.datetime.now()))
print(nwbfile)
device = nwbfile.create_device(
name='Microscope',
description='My two-photon microscope',
manufacturer='The best microscope manufacturer')
optical_channel = OpticalChannel(name='OpticalChannel',
description='an optical channel',
emission_lambda=500.)
imaging_plane = nwbfile.create_imaging_plane(
name='ImagingPlane',
optical_channel=optical_channel,
imaging_rate=ops['fs'],
description='standard',
device=device,
excitation_lambda=600.,
indicator='GCaMP',
location='V1',
grid_spacing=([2, 2, 30] if multiplane else [2, 2]),
grid_spacing_unit='microns')
# link to external data
image_series = TwoPhotonSeries(
name='TwoPhotonSeries',
dimension=[ops['Ly'], ops['Lx']],
external_file=(ops['filelist'] if 'filelist' in ops else ['']),
imaging_plane=imaging_plane,
starting_frame=[0],
format='external',
starting_time=0.0,
rate=ops['fs'] * ops['nplanes'])
nwbfile.add_acquisition(image_series)
# processing
img_seg = ImageSegmentation()
ps = img_seg.create_plane_segmentation(name='PlaneSegmentation',
description='suite2p output',
imaging_plane=imaging_plane,
reference_images=image_series)
ophys_module = nwbfile.create_processing_module(
name='ophys', description='optical physiology processed data')
ophys_module.add(img_seg)
file_strs = ['F.npy', 'Fneu.npy', 'spks.npy']
traces = []
ncells_all = 0
for iplane, ops in enumerate(ops1):
if iplane == 0:
iscell = np.load(os.path.join(ops['save_path'], 'iscell.npy'))
for fstr in file_strs:
traces.append(np.load(os.path.join(ops['save_path'],
fstr)))
else:
iscell = np.append(iscell,
np.load(
os.path.join(ops['save_path'],
'iscell.npy')),
axis=0)
for i, fstr in enumerate(file_strs):
traces[i] = np.append(
traces[i],
np.load(os.path.join(ops['save_path'], fstr)),
axis=0)
stat = np.load(os.path.join(ops['save_path'], 'stat.npy'),
allow_pickle=True)
ncells = len(stat)
for n in range(ncells):
if multiplane:
pixel_mask = np.array([
stat[n]['ypix'], stat[n]['xpix'],
iplane * np.ones(stat[n]['npix']), stat[n]['lam']
])
ps.add_roi(voxel_mask=pixel_mask.T)
else:
pixel_mask = np.array(
[stat[n]['ypix'], stat[n]['xpix'], stat[n]['lam']])
ps.add_roi(pixel_mask=pixel_mask.T)
ncells_all += ncells
ps.add_column('iscell', 'two columns - iscell & probcell', iscell)
rt_region = ps.create_roi_table_region(region=list(
np.arange(0, ncells_all)),
description='all ROIs')
# FLUORESCENCE (all are required)
file_strs = ['F.npy', 'Fneu.npy', 'spks.npy']
name_strs = ['Fluorescence', 'Neuropil', 'Deconvolved']
for i, (fstr, nstr) in enumerate(zip(file_strs, name_strs)):
roi_resp_series = RoiResponseSeries(name=nstr,
data=traces[i],
rois=rt_region,
unit='lumens',
rate=ops['fs'])
fl = Fluorescence(roi_response_series=roi_resp_series, name=nstr)
ophys_module.add(fl)
# BACKGROUNDS
# (meanImg, Vcorr and max_proj are REQUIRED)
bg_strs = ['meanImg', 'Vcorr', 'max_proj', 'meanImg_chan2']
nplanes = ops['nplanes']
for iplane in range(nplanes):
images = Images('Backgrounds_%d' % iplane)
for bstr in bg_strs:
if bstr in ops:
if bstr == 'Vcorr' or bstr == 'max_proj':
img = np.zeros((ops['Ly'], ops['Lx']), np.float32)
img[ops['yrange'][0]:ops['yrange'][-1],
ops['xrange'][0]:ops['xrange'][-1]] = ops[bstr]
else:
img = ops[bstr]
images.add_image(GrayscaleImage(name=bstr, data=img))
ophys_module.add(images)
with NWBHDF5IO(os.path.join(ops['save_path0'], 'suite2p', 'ophys.nwb'),
'w') as fio:
fio.write(nwbfile)
else:
print('pip install pynwb OR don"t use mesoscope recording')
import numpy as np
from pynwb.image import RGBAImage, RGBImage, GrayscaleImage
from pynwb.base import Images
img = Image.open('docs/source/logo.png') # an example image
# you can store an RGBA image
rgba_logo = RGBAImage(name='RGBA_logo', data=np.array(img))
# or RGB
rgb_logo = RGBImage(name='RGB_logo', data=np.array(img.convert('RGB')))
# or Grayscale. Here with the optional description and resolution specified.
gs_logo = GrayscaleImage(name='Grayscale_logo',
data=np.array(img.convert('L')),
description='Grayscale version of the NWB logo',
resolution=35.433071)
# Images is a container that accepts any of these image types
images = Images(name='logo_images', images=[rgb_logo, rgba_logo, gs_logo])
# Finally, do not forget to add the images object to the nwb file.
nwbfile.processing['behavior'].add(images)
####################
# Now we overwrite the file with all of the data
with NWBHDF5IO('example_file_path.nwb', 'w') as io:
io.write(nwbfile)
####################
def test_show_grayscale_image():
data = np.random.rand(900).reshape((30, 30))
grayscale_image = GrayscaleImage(name='test_image', data=data)
assert isinstance(show_grayscale_image(grayscale_image), plt.Figure)
def add_ophys_processing_from_suite2p(save_folder,
nwbfile,
CaImaging_timestamps,
device=None,
optical_channel=None,
imaging_plane=None,
image_series=None):
"""
adapted from suite2p/suite2p/io/nwb.py "save_nwb" function
"""
plane_folders = natsorted([
f.path for f in os.scandir(save_folder)
if f.is_dir() and f.name[:5] == 'plane'
])
ops1 = [
np.load(os.path.join(f, 'ops.npy'), allow_pickle=True).item()
for f in plane_folders
]
if len(ops1) > 1:
multiplane = True
else:
multiplane = False
ops = ops1[0]
if device is None:
device = nwbfile.create_device(
name='Microscope',
description='My two-photon microscope',
manufacturer='The best microscope manufacturer')
if optical_channel is None:
optical_channel = OpticalChannel(name='OpticalChannel',
description='an optical channel',
emission_lambda=500.)
if imaging_plane is None:
imaging_plane = nwbfile.create_imaging_plane(
name='ImagingPlane',
optical_channel=optical_channel,
imaging_rate=ops['fs'],
description='standard',
device=device,
excitation_lambda=600.,
indicator='GCaMP',
location='V1',
grid_spacing=([2, 2, 30] if multiplane else [2, 2]),
grid_spacing_unit='microns')
if image_series is None:
# link to external data
image_series = TwoPhotonSeries(
name='TwoPhotonSeries',
dimension=[ops['Ly'], ops['Lx']],
external_file=(ops['filelist'] if 'filelist' in ops else ['']),
imaging_plane=imaging_plane,
starting_frame=[0],
format='external',
starting_time=0.0,
rate=ops['fs'] * ops['nplanes'])
nwbfile.add_acquisition(image_series) # otherwise, were added
# processing
img_seg = ImageSegmentation()
ps = img_seg.create_plane_segmentation(name='PlaneSegmentation',
description='suite2p output',
imaging_plane=imaging_plane,
reference_images=image_series)
ophys_module = nwbfile.create_processing_module(
name='ophys', description='optical physiology processed data')
ophys_module.add(img_seg)
file_strs = ['F.npy', 'Fneu.npy', 'spks.npy']
traces = []
ncells_all = 0
for iplane, ops in enumerate(ops1):
if iplane == 0:
iscell = np.load(
os.path.join(save_folder, 'plane%i' % iplane, 'iscell.npy'))
for fstr in file_strs:
traces.append(
np.load(os.path.join(save_folder, 'plane%i' % iplane,
fstr)))
else:
iscell = np.append(iscell,
np.load(
os.path.join(save_folder,
'plane%i' % iplane,
'iscell.npy')),
axis=0)
for i, fstr in enumerate(file_strs):
traces[i] = np.append(
traces[i],
np.load(os.path.join(save_folder, 'plane%i' % iplane,
fstr)),
axis=0)
stat = np.load(os.path.join(save_folder, 'plane%i' % iplane,
'stat.npy'),
allow_pickle=True)
ncells = len(stat)
for n in range(ncells):
if multiplane:
pixel_mask = np.array([
stat[n]['ypix'], stat[n]['xpix'],
iplane * np.ones(stat[n]['npix']), stat[n]['lam']
])
ps.add_roi(voxel_mask=pixel_mask.T)
else:
pixel_mask = np.array(
[stat[n]['ypix'], stat[n]['xpix'], stat[n]['lam']])
ps.add_roi(pixel_mask=pixel_mask.T)
ncells_all += ncells
ps.add_column('iscell', 'two columns - iscell & probcell', iscell)
rt_region = ps.create_roi_table_region(region=list(np.arange(
0, ncells_all)),
description='all ROIs')
# FLUORESCENCE (all are required)
file_strs = ['F.npy', 'Fneu.npy', 'spks.npy']
name_strs = ['Fluorescence', 'Neuropil', 'Deconvolved']
for i, (fstr, nstr) in enumerate(zip(file_strs, name_strs)):
roi_resp_series = RoiResponseSeries(
name=nstr,
data=traces[i],
rois=rt_region,
unit='lumens',
timestamps=CaImaging_timestamps
) # CRITICAL TO HAVE IT HERE FOR RE-ALIGNEMENT
fl = Fluorescence(roi_response_series=roi_resp_series, name=nstr)
ophys_module.add(fl)
# BACKGROUNDS
# (meanImg, Vcorr and max_proj are REQUIRED)
bg_strs = ['meanImg', 'meanImgE', 'Vcorr', 'max_proj', 'meanImg_chan2']
nplanes = ops['nplanes']
for iplane in range(nplanes):
images = Images('Backgrounds_%d' % iplane)
for bstr in bg_strs:
if bstr in ops:
if bstr == 'Vcorr' or bstr == 'max_proj':
img = np.zeros((ops['Ly'], ops['Lx']), np.float32)
img[ops['yrange'][0]:ops['yrange'][-1],
ops['xrange'][0]:ops['xrange'][-1]] = ops[bstr]
else:
img = ops[bstr]
images.add_image(GrayscaleImage(name=bstr, data=img))
ophys_module.add(images)
def write_segmentation(segext_obj,
save_path,
plane_num=0,
metadata=None,
overwrite=True):
save_path = Path(save_path)
assert save_path.suffix == '.nwb'
if save_path.is_file() and not overwrite:
nwbfile_exist = True
file_mode = 'r+'
else:
if save_path.is_file():
os.remove(save_path)
if not save_path.parent.is_dir():
save_path.parent.mkdir(parents=True)
nwbfile_exist = False
file_mode = 'w'
# parse metadata correctly:
if isinstance(segext_obj, MultiSegmentationExtractor):
segext_objs = segext_obj.segmentations
if metadata is not None:
assert isinstance(metadata, list), "For MultiSegmentationExtractor enter 'metadata' as a list of " \
"SegmentationExtractor metadata"
assert len(metadata) == len(segext_objs), "The 'metadata' argument should be a list with the same " \
"number of elements as the segmentations in the " \
"MultiSegmentationExtractor"
else:
segext_objs = [segext_obj]
if metadata is not None and not isinstance(metadata, list):
metadata = [metadata]
metadata_base_list = [
NwbSegmentationExtractor.get_nwb_metadata(sgobj)
for sgobj in segext_objs
]
print(f'writing nwb for {segext_obj.extractor_name}\n')
# updating base metadata with new:
for num, data in enumerate(metadata_base_list):
metadata_input = metadata[num] if metadata else {}
metadata_base_list[num] = dict_recursive_update(
metadata_base_list[num], metadata_input)
# loop for every plane:
with NWBHDF5IO(str(save_path), file_mode) as io:
metadata_base_common = metadata_base_list[0]
if nwbfile_exist:
nwbfile = io.read()
else:
nwbfile = NWBFile(**metadata_base_common['NWBFile'])
# Subject:
if metadata_base_common.get('Subject'):
nwbfile.subject = Subject(
**metadata_base_common['Subject'])
# Processing Module:
if 'ophys' not in nwbfile.processing:
ophys = nwbfile.create_processing_module(
'ophys', 'contains optical physiology processed data')
else:
ophys = nwbfile.get_processing_module('ophys')
for plane_no_loop, (segext_obj, metadata) in enumerate(
zip(segext_objs, metadata_base_list)):
# Device:
if metadata['Ophys']['Device'][0][
'name'] not in nwbfile.devices:
nwbfile.create_device(**metadata['Ophys']['Device'][0])
# ImageSegmentation:
image_segmentation_name = 'ImageSegmentation' if plane_no_loop == 0 else f'ImageSegmentation_Plane{plane_no_loop}'
if image_segmentation_name not in ophys.data_interfaces:
image_segmentation = ImageSegmentation(
name=image_segmentation_name)
ophys.add_data_interface(image_segmentation)
else:
image_segmentation = ophys.data_interfaces.get(
image_segmentation_name)
# OpticalChannel:
optical_channels = [
OpticalChannel(**i) for i in metadata['Ophys']
['ImagingPlane'][0]['optical_channel']
]
# ImagingPlane:
image_plane_name = 'ImagingPlane' if plane_no_loop == 0 else f'ImagePlane_{plane_no_loop}'
if image_plane_name not in nwbfile.imaging_planes.keys():
input_kwargs = dict(
name=image_plane_name,
device=nwbfile.get_device(metadata_base_common['Ophys']
['Device'][0]['name']),
)
metadata['Ophys']['ImagingPlane'][0][
'optical_channel'] = optical_channels
input_kwargs.update(**metadata['Ophys']['ImagingPlane'][0])
if 'imaging_rate' in input_kwargs:
input_kwargs['imaging_rate'] = float(
input_kwargs['imaging_rate'])
imaging_plane = nwbfile.create_imaging_plane(
**input_kwargs)
else:
imaging_plane = nwbfile.imaging_planes[image_plane_name]
# PlaneSegmentation:
input_kwargs = dict(
description='output from segmenting imaging plane',
imaging_plane=imaging_plane)
ps_metadata = metadata['Ophys']['ImageSegmentation'][
'plane_segmentations'][0]
if ps_metadata[
'name'] not in image_segmentation.plane_segmentations:
input_kwargs.update(**ps_metadata)
ps = image_segmentation.create_plane_segmentation(
**input_kwargs)
ps_exist = False
else:
ps = image_segmentation.get_plane_segmentation(
ps_metadata['name'])
ps_exist = True
# ROI add:
image_masks = segext_obj.get_roi_image_masks()
roi_ids = segext_obj.get_roi_ids()
accepted_list = segext_obj.get_accepted_list()
accepted_list = [] if accepted_list is None else accepted_list
rejected_list = segext_obj.get_rejected_list()
rejected_list = [] if rejected_list is None else rejected_list
accepted_ids = [
1 if k in accepted_list else 0 for k in roi_ids
]
rejected_ids = [
1 if k in rejected_list else 0 for k in roi_ids
]
roi_locations = np.array(segext_obj.get_roi_locations()).T
if not ps_exist:
ps.add_column(
name='RoiCentroid',
description=
'x,y location of centroid of the roi in image_mask')
ps.add_column(
name='Accepted',
description=
'1 if ROi was accepted or 0 if rejected as a cell during segmentation operation'
)
ps.add_column(
name='Rejected',
description=
'1 if ROi was rejected or 0 if accepted as a cell during segmentation operation'
)
for num, row in enumerate(roi_ids):
ps.add_roi(id=row,
image_mask=image_masks[:, :, num],
RoiCentroid=roi_locations[num, :],
Accepted=accepted_ids[num],
Rejected=rejected_ids[num])
# Fluorescence Traces:
if 'Flourescence' not in ophys.data_interfaces:
fluorescence = Fluorescence()
ophys.add_data_interface(fluorescence)
else:
fluorescence = ophys.data_interfaces['Fluorescence']
roi_response_dict = segext_obj.get_traces_dict()
roi_table_region = ps.create_roi_table_region(
description=f'region for Imaging plane{plane_no_loop}',
region=list(range(segext_obj.get_num_rois())))
rate = np.float('NaN') if segext_obj.get_sampling_frequency(
) is None else segext_obj.get_sampling_frequency()
for i, j in roi_response_dict.items():
data = getattr(segext_obj, f'_roi_response_{i}')
if data is not None:
data = np.asarray(data)
trace_name = 'RoiResponseSeries' if i == 'raw' else i.capitalize(
)
trace_name = trace_name if plane_no_loop == 0 else trace_name + f'_Plane{plane_no_loop}'
input_kwargs = dict(name=trace_name,
data=data.T,
rois=roi_table_region,
rate=rate,
unit='n.a.')
if trace_name not in fluorescence.roi_response_series:
fluorescence.create_roi_response_series(
**input_kwargs)
# create Two Photon Series:
if 'TwoPhotonSeries' not in nwbfile.acquisition:
warn(
'could not find TwoPhotonSeries, using ImagingExtractor to create an nwbfile'
)
# adding images:
images_dict = segext_obj.get_images_dict()
if any([image is not None for image in images_dict.values()]):
images_name = 'SegmentationImages' if plane_no_loop == 0 else f'SegmentationImages_Plane{plane_no_loop}'
if images_name not in ophys.data_interfaces:
images = Images(images_name)
for img_name, img_no in images_dict.items():
if img_no is not None:
images.add_image(
GrayscaleImage(name=img_name, data=img_no))
ophys.add(images)
# saving NWB file:
io.write(nwbfile)
# test read
with NWBHDF5IO(str(save_path), 'r') as io:
io.read()
def add_ophys_processing_from_suite2p(save_folder, nwbfile, xml,
device=None,
optical_channel=None,
imaging_plane=None,
image_series=None):
"""
adapted from suite2p/suite2p/io/nwb.py "save_nwb" function
"""
plane_folders = natsorted([ f.path for f in os.scandir(save_folder) if f.is_dir() and f.name[:5]=='plane'])
OPS = [np.load(os.path.join(f, 'ops.npy'), allow_pickle=True).item() for f in plane_folders]
if len(OPS)>1:
multiplane, nplanes = True, len(plane_folders)
pData_folder = os.path.join(save_folder, 'combined') # processed data folder -> using the "combined output from suite2p"
else:
multiplane, nplanes = False, 1
pData_folder = os.path.join(save_folder, 'plane0') # processed data folder
# find time sampling per plane
functional_chan = ('Ch1' if len(xml['Ch1']['relativeTime'])>1 else 'Ch2') # functional channel is one of the two !!
CaImaging_timestamps = xml[functional_chan]['relativeTime']+float(xml['settings']['framePeriod'])/2.
ops = np.load(os.path.join(pData_folder, 'ops.npy'), allow_pickle=True).item()
if device is None:
device = nwbfile.create_device(
name='Microscope',
description='My two-photon microscope',
manufacturer='The best microscope manufacturer')
if optical_channel is None:
optical_channel = OpticalChannel(
name='OpticalChannel',
description='an optical channel',
emission_lambda=500.)
if imaging_plane is None:
imaging_plane = nwbfile.create_imaging_plane(
name='ImagingPlane',
optical_channel=optical_channel,
imaging_rate=ops['fs'],
description='standard',
device=device,
excitation_lambda=600.,
indicator='GCaMP',
location='V1',
grid_spacing=([2,2,30] if multiplane else [2,2]),
grid_spacing_unit='microns')
if image_series is None:
# link to external data
image_series = TwoPhotonSeries(
name='TwoPhotonSeries',
dimension=[ops['Ly'], ops['Lx']],
external_file=(ops['filelist'] if 'filelist' in ops else ['']),
imaging_plane=imaging_plane,
starting_frame=[0],
format='external',
starting_time=0.0,
rate=ops['fs'] * ops['nplanes']
)
nwbfile.add_acquisition(image_series) # otherwise, were added
# processing
img_seg = ImageSegmentation()
ps = img_seg.create_plane_segmentation(
name='PlaneSegmentation',
description='suite2p output',
imaging_plane=imaging_plane,
reference_images=image_series
)
ophys_module = nwbfile.create_processing_module(
name='ophys',
description='optical physiology processed data\n TSeries-folder=%s' % save_folder)
ophys_module.add(img_seg)
file_strs = ['F.npy', 'Fneu.npy', 'spks.npy']
traces = []
iscell = np.load(os.path.join(pData_folder, 'iscell.npy')).astype(bool)
if ops['nchannels']>1:
if os.path.isfile(os.path.join(pData_folder, 'redcell_manual.npy')):
redcell = np.load(os.path.join(pData_folder, 'redcell_manual.npy'))[iscell[:,0], :]
else:
print('\n'+30*'--')
print(' /!\ no file found for the manual labelling of red cells (generate it with the red-cell labelling GUI) /!\ ')
print(' /!\ taking the raw suit2p output with the classifier settings /!\ ')
print('\n'+30*'--')
redcell = np.load(os.path.join(pData_folder, 'redcell.npy'))[iscell[:,0], :]
for fstr in file_strs:
traces.append(np.load(os.path.join(pData_folder, fstr))[iscell[:,0], :])
stat = np.load(os.path.join(pData_folder, 'stat.npy'), allow_pickle=True)
ncells = np.sum(iscell[:,0])
plane_ID = np.zeros(ncells)
for n in np.arange(ncells):
pixel_mask = np.array([stat[iscell[:,0]][n]['ypix'], stat[iscell[:,0]][n]['xpix'],
stat[iscell[:,0]][n]['lam']])
ps.add_roi(pixel_mask=pixel_mask.T)
if 'iplane' in stat[0]:
plane_ID[n] = stat[iscell[:,0]][n]['iplane']
if ops['nchannels']>1:
ps.add_column('redcell', 'two columns - redcell & probcell', redcell)
ps.add_column('plane', 'one column - plane ID', plane_ID)
rt_region = ps.create_roi_table_region(
region=list(np.arange(0, ncells)),
description='all ROIs')
# FLUORESCENCE (all are required)
file_strs = ['F.npy', 'Fneu.npy', 'spks.npy']
name_strs = ['Fluorescence', 'Neuropil', 'Deconvolved']
for i, (fstr,nstr) in enumerate(zip(file_strs, name_strs)):
roi_resp_series = RoiResponseSeries(
name=nstr,
data=traces[i],
rois=rt_region,
unit='lumens',
timestamps=CaImaging_timestamps[::nplanes]) # ideally should be shifted for each ROI depending on the plane...
fl = Fluorescence(roi_response_series=roi_resp_series, name=nstr)
ophys_module.add(fl)
# BACKGROUNDS
# (meanImg, Vcorr and max_proj are REQUIRED)
bg_strs = ['meanImg', 'meanImgE', 'Vcorr', 'max_proj', 'meanImg_chan2']
nplanes = ops['nplanes']
for iplane in range(nplanes):
images = Images('Backgrounds_%d'%iplane)
for bstr in bg_strs:
if bstr in ops:
if bstr=='Vcorr' or bstr=='max_proj':
img = np.zeros((ops['Ly'], ops['Lx']), np.float32)
img[ops['yrange'][0]:ops['yrange'][-1],
ops['xrange'][0]:ops['xrange'][-1]] = ops[bstr]
else:
img = ops[bstr]
images.add_image(GrayscaleImage(name=bstr, data=img))
ophys_module.add(images)