def save_png(arr, save_path, figsize=(5, 5), **kwargs):
f = plt.figure(figsize=figsize)
ax = f.add_subplot(111)
ax.imshow(arr, **kwargs)
pt.save_figure_without_borders(f=f, savePath=save_path)
i = 0
for retain_cell in retain_cells:
cells[retain_cell].plot_binary_mask_border(plotAxis=ax,
color=colors[i],
borderWidth=1)
cells[retain_cell].plot_binary_mask_border(plotAxis=ax2,
color=colors[i],
borderWidth=1)
i += 1
plt.show()
# save figures
pt.save_figure_without_borders(f,
os.path.join(
save_folder,
'2P_refined_ROIs_with_background.png'),
dpi=300)
pt.save_figure_without_borders(f2,
os.path.join(
save_folder,
'2P_refined_ROIs_without_background.png'),
dpi=300)
# save h5 file
save_file = h5py.File(save_file_name, 'w')
i = 0
for retain_cell in retain_cells:
print(retain_cell, ':', cells[retain_cell].get_binary_area())
currGroup = save_file.create_group('cell' + ft.int2str(i, 4))
def run():
curr_folder = os.path.dirname(os.path.realpath(__file__))
os.chdir(curr_folder)
print('getting total mask ...')
cell_f = h5py.File('cells_refined.hdf5', 'r')
h, w = cell_f['cell0000']['roi'].attrs['dimension']
total_mask = np.zeros((h, w), dtype=np.uint8)
for cell_n, cell_grp in cell_f.items():
curr_roi = ia.WeightedROI.from_h5_group(cell_grp['roi'])
curr_mask = curr_roi.get_binary_mask()
total_mask = np.logical_or(total_mask, curr_mask)
cell_f.close()
total_mask = ni.binary_dilation(total_mask, iterations=1)
# plt.imshow(total_mask)
# plt.title('total_mask')
# plt.show()
nwb_folder = os.path.dirname(curr_folder)
nwb_fn = [f for f in os.listdir(nwb_folder) if f[-4:] == '.nwb'][0]
nwb_path = os.path.join(nwb_folder, nwb_fn)
plane_n = os.path.split(curr_folder)[1]
dset_path = 'processing/motion_correction/MotionCorrection/{}/corrected/data'.format(
plane_n)
print('downsampling movie ...')
print('\tnwb_path: {}'.format(nwb_path))
print('\tdset_path: {}'.format(dset_path))
nwb_f = h5py.File(nwb_path, 'r')
dset = nwb_f[dset_path]
print('\ttotal shape: {}'.format(dset.shape))
nwb_f.close()
mov_d = downsample_mov(nwb_path=nwb_path,
dset_path=dset_path,
dr=downsample_r)
v_min = np.amin(mov_d)
v_max = np.amax(mov_d)
print('\tshape of downsampled movie: {}'.format(mov_d.shape))
print('\n\tgenerating avi ...')
if cv2.__version__[0:3] == '3.1' or cv2.__version__[0] == '4':
codex = 'XVID'
fourcc = cv2.VideoWriter_fourcc(*codex)
out = cv2.VideoWriter('marked_mov.avi',
fourcc,
30, (frame_size * 100, frame_size * 100),
isColor=True)
elif cv2.__version__[0:6] == '2.4.11':
out = cv2.VideoWriter('marked_mov.avi',
-1,
30, (frame_size * 100, frame_size * 100),
isColor=True)
elif cv2.__version__[0:3] == '2.4':
codex = 'XVID'
fourcc = cv2.cv.CV_FOURCC(*codex)
out = cv2.VideoWriter('marked_mov.avi',
fourcc,
30, (frame_size * 100, frame_size * 100),
isColor=True)
else:
raise EnvironmentError(
'Do not understand opencv cv2 version: {}.'.format(
cv2.__version__))
f = plt.figure(figsize=(frame_size, frame_size))
for frame_i, frame in enumerate(mov_d):
print('\tframe: {} / {}'.format(frame_i, mov_d.shape[0]))
f.clear()
ax = f.add_subplot(111)
ax.imshow(frame,
vmin=v_min,
vmax=v_max * 0.5,
cmap='gray',
interpolation='nearest')
pt.plot_mask_borders(total_mask,
plotAxis=ax,
color='#ff0000',
zoom=1,
borderWidth=1)
ax.set_aspect('equal')
# plt.show()
buffer_ = io.BytesIO()
pt.save_figure_without_borders(f, buffer_, dpi=100)
buffer_.seek(0)
image = PIL.Image.open(buffer_)
curr_frame = np.asarray(image)
r, g, b, a = np.rollaxis(curr_frame, axis=-1)
curr_frame = (np.dstack((b, g, r)))
# print(r.dtype)
# print(curr_frame.shape)
out.write(curr_frame)
out.release()
cv2.destroyAllWindows()
print('\t.avi movie generated.')
def run():
# pixels, masks with center location within this pixel region at the image border will be discarded
center_margin = [
10, 30, 35, 10
] # [top margin, bottom margin, left margin, right margin]
# area range, range of number of pixels of a valid roi
area_range = [5, 500] # [10, 100] for bouton, [150, 1000] for soma
# for the two masks that are overlapping, if the ratio between overlap and the area of the smaller mask is larger than
# this value, the smaller mask will be discarded.
overlap_thr = 0 # 0.2
save_folder = 'figures'
data_file_name = 'cells.hdf5'
save_file_name = 'cells_refined.hdf5'
background_file_name = "corrected_mean_projections.tif"
curr_folder = os.path.dirname(os.path.realpath(__file__))
os.chdir(curr_folder)
if not os.path.isdir(save_folder):
os.makedirs(save_folder)
# read cells
dfile = h5py.File(data_file_name, 'r')
cells = {}
for cellname in dfile.keys():
cells.update({cellname: ia.WeightedROI.from_h5_group(dfile[cellname])})
print('total number of cells:', len(cells))
# get the names of cells which are on the edge
edge_cells = []
for cellname, cellmask in cells.items():
dimension = cellmask.dimension
center = cellmask.get_center()
if center[0] < center_margin[0] or \
center[0] > dimension[0] - center_margin[1] or \
center[1] < center_margin[2] or \
center[1] > dimension[1] - center_margin[3]:
# cellmask.plot_binary_mask_border(color='#ff0000', borderWidth=1)
# plt.title(cellname)
# plt.show()
edge_cells.append(cellname)
print('\ncells to be removed because they are on the edges:')
print('\n'.join(edge_cells))
# remove edge cells
for edge_cell in edge_cells:
_ = cells.pop(edge_cell)
# get dictionary of cell areas
cell_areas = {}
for cellname, cellmask in cells.items():
cell_areas.update({cellname: cellmask.get_binary_area()})
# remove cellnames that have area outside of the area_range
invalid_cell_ns = []
for cellname, cellarea in cell_areas.items():
if cellarea < area_range[0] or cellarea > area_range[1]:
invalid_cell_ns.append(cellname)
print("cells to be removed because they do not meet area criterion:")
print("\n".join(invalid_cell_ns))
for invalid_cell_n in invalid_cell_ns:
cell_areas.pop(invalid_cell_n)
# sort cells with their binary area
cell_areas_sorted = sorted(cell_areas.items(), key=operator.itemgetter(1))
cell_areas_sorted.reverse()
cell_names_sorted = [c[0] for c in cell_areas_sorted]
# print '\n'.join([str(c) for c in cell_areas_sorted])
# get the name of cells that needs to be removed because of overlapping
retain_cells = []
remove_cells = []
for cell1_name in cell_names_sorted:
cell1_mask = cells[cell1_name]
is_remove = 0
cell1_area = cell1_mask.get_binary_area()
for cell2_name in retain_cells:
cell2_mask = cells[cell2_name]
cell2_area = cell2_mask.get_binary_area()
curr_overlap = cell1_mask.binary_overlap(cell2_mask)
if float(curr_overlap) / cell1_area > overlap_thr:
remove_cells.append(cell1_name)
is_remove = 1
print(cell1_name, ':', cell1_mask.get_binary_area(),
': removed')
# f = plt.figure(figsize=(10,10))
# ax = f.add_subplot(111)
# cell1_mask.plot_binary_mask_border(plotAxis=ax, color='#ff0000', borderWidth=1)
# cell2_mask.plot_binary_mask_border(plotAxis=ax, color='#0000ff', borderWidth=1)
# ax.set_title('red:'+cell1_name+'; blue:'+cell2_name)
# plt.show()
break
if is_remove == 0:
retain_cells.append(cell1_name)
print(cell1_name, ':', cell1_mask.get_binary_area(), ': retained')
print('\ncells to be removed because of overlapping:')
print('\n'.join(remove_cells))
print('\ntotal number of reatined cells:', len(retain_cells))
# plotting
colors = pt.random_color(len(cells.keys()))
bgImg = ia.array_nor(np.max(tf.imread(background_file_name), axis=0))
f = plt.figure(figsize=(10, 10))
ax = f.add_subplot(111)
ax.imshow(ia.array_nor(bgImg),
cmap='gray',
vmin=0,
vmax=0.5,
interpolation='nearest')
f2 = plt.figure(figsize=(10, 10))
ax2 = f2.add_subplot(111)
ax2.imshow(np.zeros(bgImg.shape, dtype=np.uint8),
vmin=0,
vmax=1,
cmap='gray',
interpolation='nearest')
i = 0
for retain_cell in retain_cells:
cells[retain_cell].plot_binary_mask_border(plotAxis=ax,
color=colors[i],
borderWidth=1)
cells[retain_cell].plot_binary_mask_border(plotAxis=ax2,
color=colors[i],
borderWidth=1)
i += 1
# plt.show()
# save figures
pt.save_figure_without_borders(f,
os.path.join(
save_folder,
'2P_refined_ROIs_with_background.png'),
dpi=300)
pt.save_figure_without_borders(
f2,
os.path.join(save_folder, '2P_refined_ROIs_without_background.png'),
dpi=300)
# save h5 file
save_file = h5py.File(save_file_name, 'x')
i = 0
for retain_cell in retain_cells:
print(retain_cell, ':', cells[retain_cell].get_binary_area())
currGroup = save_file.create_group('cell' + ft.int2str(i, 4))
currGroup.attrs['name'] = retain_cell
roiGroup = currGroup.create_group('roi')
cells[retain_cell].to_h5_group(roiGroup)
i += 1
for attr, value in dfile.attrs.items():
save_file.attrs[attr] = value
save_file.close()
dfile.close()
def run():
data_file_name = 'cells_refined.hdf5'
background_file_name = "corrected_mean_projections.tif"
save_folder = 'figures'
overlap_threshold = 0.9
surround_limit = [1, 8]
curr_folder = os.path.dirname(os.path.realpath(__file__))
os.chdir(curr_folder)
if not os.path.isdir(save_folder):
os.makedirs(save_folder)
print('reading cells file ...')
data_f = h5py.File(data_file_name, 'r')
cell_ns = data_f.keys()
cell_ns.sort()
binary_mask_array = []
weight_mask_array = []
for cell_n in cell_ns:
curr_roi = ia.ROI.from_h5_group(data_f[cell_n]['roi'])
binary_mask_array.append(curr_roi.get_binary_mask())
weight_mask_array.append(curr_roi.get_weighted_mask())
data_f.close()
binary_mask_array = np.array(binary_mask_array)
weight_mask_array = np.array(weight_mask_array)
print('starting mask_array shape:', weight_mask_array.shape)
print('getting total mask ...')
total_mask = np.zeros((binary_mask_array.shape[1], binary_mask_array.shape[2]), dtype=np.uint8)
for curr_mask in binary_mask_array:
total_mask = np.logical_or(total_mask, curr_mask)
total_mask = np.logical_not(total_mask)
plt.imshow(total_mask, interpolation='nearest')
plt.title('total_mask')
# plt.show()
print('getting and surround masks ...')
binary_surround_array = []
for binary_center in binary_mask_array:
curr_surround = np.logical_xor(ni.binary_dilation(binary_center, iterations=surround_limit[1]),
ni.binary_dilation(binary_center, iterations=surround_limit[0]))
curr_surround = np.logical_and(curr_surround, total_mask).astype(np.uint8)
binary_surround_array.append(curr_surround)
# plt.imshow(curr_surround)
# plt.show()
binary_surround_array = np.array(binary_surround_array)
print("saving rois ...")
center_areas = []
surround_areas = []
for mask_ind in range(binary_mask_array.shape[0]):
center_areas.append(np.sum(binary_mask_array[mask_ind].flat))
surround_areas.append(np.sum(binary_surround_array[mask_ind].flat))
roi_f = h5py.File('rois_and_traces.hdf5')
roi_f['masks_center'] = weight_mask_array
roi_f['masks_surround'] = binary_surround_array
roi_f.close()
print('minimum surround area:', min(surround_areas), 'pixels.')
f = plt.figure(figsize=(10, 10))
ax_center = f.add_subplot(211)
ax_center.hist(center_areas, bins=30)
ax_center.set_title('roi center area distribution')
ax_surround = f.add_subplot(212)
ax_surround.hist(surround_areas, bins=30)
ax_surround.set_title('roi surround area distribution')
# plt.show()
print('plotting ...')
colors = pt.random_color(weight_mask_array.shape[0])
bg = ia.array_nor(np.max(tf.imread(background_file_name), axis=0))
f_c_bg = plt.figure(figsize=(10, 10))
ax_c_bg = f_c_bg.add_subplot(111)
ax_c_bg.imshow(bg, cmap='gray', vmin=0, vmax=0.5, interpolation='nearest')
f_c_nbg = plt.figure(figsize=(10, 10))
ax_c_nbg = f_c_nbg.add_subplot(111)
ax_c_nbg.imshow(np.zeros(bg.shape,dtype=np.uint8),vmin=0,vmax=1,cmap='gray',interpolation='nearest')
f_s_nbg = plt.figure(figsize=(10, 10))
ax_s_nbg = f_s_nbg.add_subplot(111)
ax_s_nbg.imshow(np.zeros(bg.shape,dtype=np.uint8),vmin=0,vmax=1,cmap='gray',interpolation='nearest')
i = 0
for mask_ind in range(binary_mask_array.shape[0]):
pt.plot_mask_borders(binary_mask_array[mask_ind], plotAxis=ax_c_bg, color=colors[i], borderWidth=1)
pt.plot_mask_borders(binary_mask_array[mask_ind], plotAxis=ax_c_nbg, color=colors[i], borderWidth=1)
pt.plot_mask_borders(binary_surround_array[mask_ind], plotAxis=ax_s_nbg, color=colors[i], borderWidth=1)
i += 1
# plt.show()
print('saving figures ...')
pt.save_figure_without_borders(f_c_bg, os.path.join(save_folder, '2P_ROIs_with_background.png'), dpi=300)
pt.save_figure_without_borders(f_c_nbg, os.path.join(save_folder, '2P_ROIs_without_background.png'), dpi=300)
pt.save_figure_without_borders(f_s_nbg, os.path.join(save_folder, '2P_ROI_surrounds_background.png'), dpi=300)
f.savefig(os.path.join(save_folder, 'roi_area_distribution.pdf'), dpi=300)