def qc_background_regions(intensity_imp, bg_rois):
"""allow the user to view and correct automatically-determined background regions"""
imp = Duplicator().run(intensity_imp);
imp.setTitle("Background region QC");
imp.show();
imp.setPosition(1);
autoset_zoom(imp);
imp.setRoi(bg_rois[0]);
IJ.setTool("freehand");
notOK = True;
while notOK:
listener = UpdateRoiImageListener(bg_rois, is_area=True);
imp.addImageListener(listener);
dialog = NonBlockingGenericDialog("Background region quality control");
dialog.enableYesNoCancel("Continue", "Use this region for all t");
dialog.setCancelLabel("Cancel analysis");
dialog.addMessage("Please redraw background regions as necessary...")
dialog.showDialog();
if dialog.wasCanceled():
raise KeyboardInterrupt("Run canceled");
elif not(dialog.wasOKed()):
this_roi = imp.getRoi();
bg_rois = [this_roi for _ in listener.getRoiList()];
imp.removeImageListener(listener);
else:
last_roi = imp.getRoi();
qcd_bg_rois = listener.getRoiList();
if imp.getNFrames() > imp.getNSlices():
qcd_bg_rois[imp.getT() - 1] = last_roi;
else:
qcd_bg_rois[imp.getZ() - 1] = last_roi;
notOK = False;
imp.removeImageListener(listener);
imp.changes = False;
imp.close();
return qcd_bg_rois;
def generate_background_rois(input_mask_imp,
params,
membrane_edges,
dilations=5,
threshold_method=None,
membrane_imp=None):
"""automatically identify background region based on auto-thresholded image, existing membrane edges and position of midpoint anchor"""
if input_mask_imp is None and membrane_imp is not None:
segmentation_imp = Duplicator().run(membrane_imp)
# do thresholding using either previous method if threhsold_method is None or using (less conservative?) threshold method
if (threshold_method is None
or not (threshold_method in params.listThresholdMethods())):
mask_imp = make_and_clean_binary(segmentation_imp,
params.threshold_method)
else:
mask_imp = make_and_clean_binary(segmentation_imp,
threshold_method)
segmentation_imp.close()
else:
input_mask_imp.killRoi()
mask_imp = Duplicator().run(input_mask_imp)
rois = []
IJ.setForegroundColor(0, 0, 0)
roim = RoiManager(True)
rt = ResultsTable()
for fridx in range(mask_imp.getNFrames()):
mask_imp.setT(fridx + 1)
# add extra bit to binary mask from loaded membrane in case user refined edges...
# flip midpoint anchor across the line joining the two extremes of the membrane,
# and fill in the triangle made by this new point and those extremes
poly = membrane_edges[fridx].getPolygon()
l1 = (poly.xpoints[0], poly.ypoints[0])
l2 = (poly.xpoints[-1], poly.ypoints[-1])
M = (0.5 * (l1[0] + l2[0]), 0.5 * (l1[1] + l2[1]))
Mp1 = (params.manual_anchor_midpoint[0][0] - M[0],
params.manual_anchor_midpoint[0][1] - M[1])
p2 = (M[0] - Mp1[0], M[1] - Mp1[1])
new_poly_x = list(poly.xpoints)
new_poly_x.append(p2[0])
new_poly_y = list(poly.ypoints)
new_poly_y.append(p2[1])
mask_imp.setRoi(PolygonRoi(new_poly_x, new_poly_y, PolygonRoi.POLYGON))
IJ.run(mask_imp, "Fill", "slice")
mask_imp.killRoi()
# now dilate the masked image and identify the unmasked region closest to the midpoint anchor
ip = mask_imp.getProcessor()
dilations = 5
for d in range(dilations):
ip.dilate()
ip.invert()
mask_imp.setProcessor(ip)
mxsz = mask_imp.getWidth() * mask_imp.getHeight()
pa = ParticleAnalyzer(
ParticleAnalyzer.ADD_TO_MANAGER | ParticleAnalyzer.SHOW_PROGRESS,
ParticleAnalyzer.CENTROID, rt, 0, mxsz)
pa.setRoiManager(roim)
pa.analyze(mask_imp)
ds_to_anchor = [
math.sqrt((x - params.manual_anchor_midpoint[0][0])**2 +
(y - params.manual_anchor_midpoint[0][1])**2)
for x, y in zip(
rt.getColumn(rt.getColumnIndex("X")).tolist(),
rt.getColumn(rt.getColumnIndex("Y")).tolist())
]
if len(ds_to_anchor) > 0:
roi = roim.getRoi(ds_to_anchor.index(min(ds_to_anchor)))
rois.append(roi)
else:
rois.append(None)
roim.reset()
rt.reset()
roim.close()
mask_imp.close()
return rois
def ColMigBud():
def setupDialog(imp):
gd = GenericDialog("Collective migration buddy options")
gd.addMessage("Collective migration buddy 2.0, you are analyzing: " +
imp.getTitle())
calibration = imp.getCalibration()
if (calibration.frameInterval > 0):
default_interval = calibration.frameInterval
default_timeunit = calibration.getTimeUnit()
else:
default_interval = 8
default_timeunit = "min"
gd.addNumericField("Frame interval:", default_interval,
2) # show 2 decimals
gd.addCheckbox("Do you want to use a gliding window?", True)
gd.addCheckbox(
"Project hyperStack? (defaluts to projecting current channel only)",
False)
gd.addStringField("time unit", default_timeunit, 3)
gd.addSlider("Start compacting at frame:", 1, imp.getNFrames(), 1)
gd.addSlider("Stop compacting at frame:", 1, imp.getNFrames(),
imp.getNFrames())
gd.addNumericField("Number of frames to project in to one:", 3,
0) # show 0 decimals
gd.addChoice('Method to use for frame projection:', methods_as_strings,
methods_as_strings[5])
gd.showDialog()
if gd.wasCanceled():
IJ.log("User canceled dialog!")
return
return gd
#Start by getting the active image window and get the current active channel and other stats
imp = WindowManager.getCurrentImage()
cal = imp.getCalibration()
nSlices = 1 #TODO fix this in case you want to do Z-stacks
title = imp.getTitle()
current_channel = imp.getChannel()
#zp = ZProjector(imp)
#Make a dict containg method_name:const_fieled_value pairs for the projection methods
methods_as_strings = [
'Average Intensity', 'Max Intensity', 'Min Intensity', 'Sum Slices',
'Standard Deviation', 'Median'
]
methods_as_const = [
ZProjector.AVG_METHOD, ZProjector.MAX_METHOD, ZProjector.MIN_METHOD,
ZProjector.SUM_METHOD, ZProjector.SD_METHOD, ZProjector.MEDIAN_METHOD
]
medthod_dict = dict(zip(methods_as_strings, methods_as_const))
# Run the setupDialog, read out and store the options
gd = setupDialog(imp)
frame_interval = gd.getNextNumber()
time_unit = gd.getNextString()
glidingFlag = gd.getNextBoolean()
hyperstackFlag = gd.getNextBoolean()
#Set the frame interval and unit, and store it in the ImagePlus calibration
cal.frameInterval = frame_interval
cal.setTimeUnit(time_unit)
imp.setCalibration(cal)
start_frame = int(gd.getNextNumber())
stop_frame = int(gd.getNextNumber())
#If a subset of the image is to be projected, these lines of code handle that
if (start_frame > stop_frame):
IJ.showMessage("Start frame > Stop frame, can't go backwards in time!")
raise RuntimeException("Start frame > Stop frame!")
if ((start_frame != 1) or (stop_frame != imp.getNFrames())):
imp = Duplicator().run(imp, 1, nChannels, 1, nSlices, start_frame,
stop_frame)
no_frames_per_integral = int(gd.getNextNumber())
#the doHyperstackProjection method can't project past the end of the stack
if hyperstackFlag:
total_no_frames_to_project = imp.getNFrames() - no_frames_per_integral
#the doProjection method can project past the end, it just adds black frames at the end
#When not projecting hyperstacks, just copy the current active channel from the active image
else:
total_no_frames_to_project = imp.getNFrames()
imp = Duplicator().run(imp, current_channel, current_channel, 1,
nSlices, start_frame, stop_frame)
#The Z-Projection magic happens here through a ZProjector object
zp = ZProjector(imp)
projection_method = gd.getNextChoice()
chosen_method = medthod_dict[projection_method]
zp.setMethod(chosen_method)
outstack = imp.createEmptyStack()
if glidingFlag:
frames_to_advance_per_step = 1
else:
frames_to_advance_per_step = no_frames_per_integral
for frame in range(1, total_no_frames_to_project,
frames_to_advance_per_step):
zp.setStartSlice(frame)
zp.setStopSlice(frame + no_frames_per_integral)
if hyperstackFlag:
zp.doHyperStackProjection(False)
projected_stack = zp.getProjection().getStack()
for channel in range(projected_stack.getSize()):
outstack.addSlice(projected_stack.getProcessor(channel + 1))
else:
zp.doProjection()
outstack.addSlice(zp.getProjection().getProcessor())
#Create an image processor from the newly created Z-projection stack
nChannels = imp.getNChannels()
nFrames = outstack.getSize() / nChannels
imp2 = ImagePlus(
title + '_' + projection_method + '_' + str(no_frames_per_integral) +
'_frames', outstack)
imp2 = HyperStackConverter.toHyperStack(imp2, nChannels, nSlices, nFrames)
imp2.show()
Startmenu()
