def do_tubefitting(im_path=im_test_path,
metadata_path=metadata_test_path,
output_path=output_path,
save_output=False):
# todo: fix things so that all operations use a consistent definition of background rather than changing Prefs on the fly...
Prefs.blackBackground = False
info = PrescreenInfo()
info.load_info_from_json(metadata_path)
z_xy_ratio = abs(
info.get_z_plane_spacing_um()) / info.get_xy_pixel_size_um()
#z_xy_ratio = 1.0;
bfimp = bf.openImagePlus(im_path)
imp = bfimp[0]
imp.show()
IJ.run(imp, "Set Scale...", "distance=0 known=0 pixel=1 unit=pixel")
imp = utils.downsample_for_isotropy(imp,
extra_downsample_factor=1.0,
info=info)
rot_seg_imp, rot_proj_imp, egfp_mch_imps = split_and_rotate(imp, info)
depth = rot_seg_imp.getNSlices() if rot_seg_imp.getNSlices(
) > rot_seg_imp.getNFrames() else rot_seg_imp.getNFrames()
width = rot_seg_imp.getWidth()
height = int(round(rot_seg_imp.getHeight() * z_xy_ratio))
# Apply 3d MEDIAN FILTER to denoise and emphasise vessel-associated voxels
fit_basis_imp = threshold_and_binarise(rot_seg_imp, z_xy_ratio)
fit_basis_imp.setTitle("fit_basis_imp")
fit_basis_imp.show()
# plane-wise, use binary-outline
# say the non-zero points then make up basis for fitting to be performed per http://nicky.vanforeest.com/misc/fitEllipse/fitEllipse.html
rois = []
centres = []
major_axes = []
roi_imp = IJ.createImage("rois", width, height, depth, 32)
pts_stack = ImageStack(width, height + 1)
IJ.run(imp, "Line Width...", "line=3")
for zidx in range(fit_basis_imp.getNSlices()):
fit_basis_imp.setZ(zidx + 1)
IJ.run(fit_basis_imp, "Outline", "slice")
IJ.run(fit_basis_imp, "Create Selection", "")
roi = fit_basis_imp.getRoi()
fit_basis_imp.killRoi()
pts = [(pt.x, pt.y) for pt in roi.getContainedPoints()]
clean_pts = convex_hull_pts(pts)
clean_pts = [(x, z_xy_ratio * y) for (x, y) in clean_pts]
# make a stack of clean points...
ip = FloatProcessor(width, height + 1)
pix = ip.getPixels()
for pt in clean_pts:
pix[int(pt[1]) * width + int(pt[0])] = 128
pts_stack.addSlice(ip)
centre, angle, axl = ellipse_fitting.fit_ellipse(clean_pts)
major_axes.append(max(axl))
centres.append(centre)
rot_seg_imp.setZ(zidx + 1)
ellipse_roi = ellipse_fitting.generate_ellipse_roi(centre, angle, axl)
rois.append(ellipse_roi)
IJ.run(imp, "Line Width...", "line=1")
cal = imp.getCalibration()
smooth_centres, tangent_vecs = generate_smoothed_vessel_axis(
centres, pixel_size_um=cal.pixelDepth)
for zidx in range(fit_basis_imp.getNSlices()):
centre = smooth_centres[zidx]
major_axis = major_axes[zidx]
ellipse_roi = EllipseRoi(centre[0] - 2, centre[1], centre[0] + 2,
centre[1], 1.0)
roi_imp.setZ(zidx + 1)
roi_imp.setRoi(ellipse_roi)
IJ.run(roi_imp, "Set...",
"value=" + str(roi_imp.getProcessor().maxValue()) + " slice")
pts_stack_imp = ImagePlus("Cleaned points", pts_stack)
pts_stack_imp.setTitle("pts_stack_imp")
pts_stack_imp.show()
rot_seg_imp.changes = False
rot_seg_imp.close()
egfp_imp = egfp_mch_imps[0]
mch_imp = egfp_mch_imps[1]
imps_to_combine = [egfp_mch_imps[1], egfp_mch_imps[0], roi_imp]
egfp_imp.show()
mch_imp.show()
roi_imp.show()
print("box height um = " +
str(roi_imp.getNSlices() * info.get_xy_pixel_size_um()))
IJ.run(
egfp_imp, "Size...", "width=" + str(width) + " height=" + str(height) +
" depth=" + str(depth) + " average interpolation=Bilinear")
IJ.run(
mch_imp, "Size...", "width=" + str(width) + " height=" + str(height) +
" depth=" + str(depth) + " average interpolation=Bilinear")
#IJ.run("Merge Channels...", "c1=[" + mch_imp.getTitle() +
# "] c2=[" + egfp_imp.getTitle() +
# "] c7=[" + roi_imp.getTitle() + "] create keep");
composite_imp = RGBStackMerge().mergeChannels(imps_to_combine, False)
print(composite_imp)
composite_imp.show()
print("end of vessel centerline id step, image dims = ({}x{}x{})".format(
composite_imp.getWidth(), composite_imp.getHeight(),
composite_imp.getNSlices()))
WaitForUserDialog("pause").show()
# do qc here?
#WM.getImage("Composite").addImageListener(UpdateRoiImageListener(rois));
IJ.run(roi_imp, "8-bit", "")
if save_output:
FileSaver(composite_imp).saveAsTiffStack(
os.path.join(output_path, "segmentation result.tif"))
print(roi_imp)
FileSaver(roi_imp).saveAsTiff(
os.path.join(output_path, "vessel axis.tif"))
egfp_imp.changes = False
mch_imp.changes = False
roi_imp.changes = False
fit_basis_imp.changes = False
pts_stack_imp.changes = False
egfp_imp.close()
mch_imp.close()
#roi_imp.close();
fit_basis_imp.close()
pts_stack_imp.close()
zcoords = [i for i in range(composite_imp.getNSlices())]
xyz_smooth_centres = [(x, y, z)
for ((x, y), z) in zip(smooth_centres, zcoords)]
composite_imp2 = straighten_vessel(composite_imp,
xyz_smooth_centres,
save_output=True)
composite_imp3 = straighten_vessel(composite_imp2,
xyz_smooth_centres,
it=2,
save_output=True)
return composite_imp3
def downsample_for_isotropy(imp, extra_downsample_factor=2.0, info=None):
"""downsample x, y pixel directions to get ~cubic voxels"""
title = imp.getTitle();
cal = imp.getCalibration();
if info is None:
pix_w = cal.pixelWidth;
pix_h = cal.pixelHeight;
pix_d = cal.pixelDepth;
else:
pix_w = info.get_xy_pixel_size_um();
pix_h = pix_w;
pix_d = info.get_z_plane_spacing_um();
im_w = imp.getWidth();
im_h = imp.getHeight();
im_d = imp.getNSlices();
print("original pixel whd = ({}, {}, {})".format(pix_w, pix_h, pix_d));
print("original image whd = ({}, {}, {})".format(im_w, im_h, im_d));
im_nch = imp.getNChannels();
if im_nch > 1:
split_ch = ChannelSplitter().split(imp);
else:
split_ch = [imp];
print("downsampling {} and making isotropic...".format(title));
IJ.showStatus("Downsampling and making ~isotropic...");
xy_scale = pix_h / (pix_d * extra_downsample_factor);
xy_scaled_h = int(xy_scale * im_h);
xy_scaled_w = int(xy_scale * im_w);
z_scale = 1/ extra_downsample_factor;
z_scaled_h = int(z_scale * im_d);
out_imps = [];
for ch_imp in split_ch:
print(ch_imp.getTitle());
sp = StackProcessor(ch_imp.getStack());
print((xy_scaled_w, xy_scaled_h));
stack = sp.resize(xy_scaled_w, xy_scaled_h, True);
xz_stack = rot_around_x(stack);
xz_sp = StackProcessor(xz_stack);
xz_stack = xz_sp.resize(xy_scaled_w, z_scaled_h, True);
out_stack = rot_around_x(xz_stack);
out_imps.append(ImagePlus("Isotropic downsampled {}".format(title), out_stack));
cal.setUnit('um');
cal.pixelWidth = im_w/xy_scaled_w * pix_w;
cal.pixelHeight = im_h/xy_scaled_h * pix_h;
cal.pixelDepth = im_d/z_scaled_h * pix_d;
print("new pixel whd = ({}, {}, {})".format(cal.pixelWidth, cal.pixelHeight, cal.pixelDepth));
imp.changes = False;
imp.close();
for ch_imp in split_ch:
ch_imp.close();
if len(out_imps) > 1:
out_imp = RGBStackMerge().mergeChannels(out_imps, False);
else:
out_imp = out_imps[0];
out_imp.setCalibration(cal);
print("new image whd = ({}, {}, {})".format(out_imp.getWidth(), out_imp.getHeight(), out_imp.getNSlices()));
print("...done downsampling {} and making isotropic. ".format(title));
IJ.showStatus("...done downsampling and making ~isotropic. ");
return out_imp;
