def trackmate(self):
calibration = self.imp.getCalibration()
model = Model()
model.setLogger(Logger.IJ_LOGGER)
settings = Settings()
settings.setFrom(self.imp)
# Configure detector - We use the Strings for the keys
settings.detectorFactory = LogDetectorFactory()
settings.detectorSettings = {
'DO_SUBPIXEL_LOCALIZATION': True,
'RADIUS': calibration.getX(self.trackmateSize),
'TARGET_CHANNEL': 1,
'THRESHOLD': self.trackmateThreashold,
'DO_MEDIAN_FILTERING': True,
}
# Configure spot filters - Classical filter on quality
filter1 = FeatureFilter('QUALITY', 0.01, True)
settings.addSpotFilter(filter1)
settings.addSpotAnalyzerFactory(SpotIntensityMultiCAnalyzerFactory())
settings.initialSpotFilterValue = 1
# Configure tracker - We want to allow merges and fusions
settings.trackerFactory = SparseLAPTrackerFactory()
settings.trackerSettings = LAPUtils.getDefaultLAPSettingsMap()
trackmate = TrackMate(model, settings)
#--------
# Process
#--------
ok = trackmate.checkInput()
if not ok:
print("NOT OK")
ok = trackmate.process()
if not ok:
print("NOT OK")
#----------------
# Display results
#----------------
#selectionModel = SelectionModel(model)
#displayer = HyperStackDisplayer(model, selectionModel, self.imp)
#displayer.render()
#displayer.refresh()
# Echo results with the logger we set at start:
spots = model.getSpots()
return spots.iterable(True)
def runTrackMate(imp, targetChannel, dt, radius, threshold, frameGap,
linkingMax, closingMax):
# Get the number of channels
nChannels = imp.getNChannels()
IJ.log("->Detection threshold used: " + str(threshold))
IJ.log("->Number of frames is: " + str(imp.getStackSize()))
IJ.log("->Target channel is: " + str(targetChannel))
IJ.log('->Number of channels to measure %d' % nChannels)
# Setup settings for TrackMate
settings = Settings()
settings.setFrom(imp)
settings.dt = dt
# Spot analyzer: we want the multi-C intensity analyzer.
settings.addSpotAnalyzerFactory(SpotMultiChannelIntensityAnalyzerFactory())
# Spot detector.
settings.detectorFactory = LogDetectorFactory()
settings.detectorSettings = settings.detectorFactory.getDefaultSettings()
settings.detectorSettings['RADIUS'] = radius
settings.detectorSettings['THRESHOLD'] = threshold
settings.detectorSettings['TARGET_CHANNEL'] = targetChannel
# Spot tracker.
#settings.trackerFactory = SparseLAPTrackerFactory()
settings.trackerFactory = LAPTrackerFactory()
settings.trackerSettings = LAPUtils.getDefaultLAPSettingsMap()
settings.trackerSettings['MAX_FRAME_GAP'] = frameGap
settings.trackerSettings['LINKING_MAX_DISTANCE'] = linkingMax
settings.trackerSettings['GAP_CLOSING_MAX_DISTANCE'] = closingMax
settings.trackerSettings['ALLOW_TRACK_MERGING'] = False
settings.trackerSettings['ALLOW_GAP_CLOSING'] = False
settings.trackerSettings['ALLOW_TRACK_SPLITTING'] = False
settings.trackerSettings['ALLOW_TRACK_MERGING'] = False
settings.trackerSettings['ALTERNATIVE_LINKING_COST_FACTOR'] = 0.5
settings.trackerSettings['BLOCKING_VALUE'] = 1.0
settings.trackerSettings['CUTOFF_PERCENTILE'] = 1.0
#settings.trackerSettings['SPLITTING_MAX_DISTANCE'] = 16.0
# Run TrackMate and store data into Model.
model = Model()
trackmate = TrackMate(model, settings)
if not trackmate.checkInput() or not trackmate.process():
IJ.log('Could not execute TrackMate: ' +
str(trackmate.getErrorMessage()))
else:
return model, nChannels
# empty model now.
model = Model()
# Send all messages to ImageJ log window.
model.setLogger(Logger.IJ_LOGGER)
#------------------------
# Prepare settings object
#------------------------
settings = Settings()
settings.setFrom(imp)
# Configure detector - We use the Strings for the keys
settings.detectorFactory = LogDetectorFactory()
settings.detectorSettings = {
'DO_SUBPIXEL_LOCALIZATION': True,
'RADIUS': 2.5,
'TARGET_CHANNEL': 1,
'THRESHOLD': 0.,
'DO_MEDIAN_FILTERING': False,
}
# Configure spot filters - Classical filter on quality
filter1 = FeatureFilter('QUALITY', 1, True)
settings.addSpotFilter(filter1)
# Configure tracker - We want to allow merges and fusions
settings.trackerFactory = SparseLAPTrackerFactory()
settings.trackerSettings = LAPUtils.getDefaultLAPSettingsMap(
def track(imp):
from fiji.plugin.trackmate import Model
from fiji.plugin.trackmate import Settings
from fiji.plugin.trackmate import TrackMate
from fiji.plugin.trackmate import SelectionModel
from fiji.plugin.trackmate import Logger
from fiji.plugin.trackmate.detection import LogDetectorFactory
from fiji.plugin.trackmate.tracking.sparselap import SparseLAPTrackerFactory
from fiji.plugin.trackmate.tracking import LAPUtils
from ij import IJ
import fiji.plugin.trackmate.visualization.hyperstack.HyperStackDisplayer as HyperStackDisplayer
import fiji.plugin.trackmate.features.FeatureFilter as FeatureFilter
import sys
import fiji.plugin.trackmate.features.track.TrackDurationAnalyzer as TrackDurationAnalyzer
# Get currently selected image
#imp = WindowManager.getCurrentImage()
#imp = IJ.openImage('http://fiji.sc/samples/FakeTracks.tif')
#imp.show()
#----------------------------
# Create the model object now
#----------------------------
# Some of the parameters we configure below need to have
# a reference to the model at creation. So we create an
# empty model now.
model = Model()
# Send all messages to ImageJ log window.
model.setLogger(Logger.IJ_LOGGER)
#------------------------
# Prepare settings object
#------------------------
settings = Settings()
settings.setFrom(imp)
# Configure detector - We use the Strings for the keys
settings.detectorFactory = LogDetectorFactory()
settings.detectorSettings = {
'DO_SUBPIXEL_LOCALIZATION': True,
'RADIUS': 3.0,
'TARGET_CHANNEL': 1,
'THRESHOLD': 1.,
'DO_MEDIAN_FILTERING': False,
}
# Configure spot filters - Classical filter on quality
filter1 = FeatureFilter('QUALITY', 30, True)
settings.addSpotFilter(filter1)
# Configure tracker - We want to allow merges and fusions
settings.trackerFactory = SparseLAPTrackerFactory()
settings.trackerSettings = LAPUtils.getDefaultLAPSettingsMap(
) # almost good enough
settings.trackerSettings['ALLOW_TRACK_SPLITTING'] = True
settings.trackerSettings['ALLOW_TRACK_MERGING'] = True
# Configure track analyzers - Later on we want to filter out tracks
# based on their displacement, so we need to state that we want
# track displacement to be calculated. By default, out of the GUI,
# not features are calculated.
# The displacement feature is provided by the TrackDurationAnalyzer.
settings.addTrackAnalyzer(TrackDurationAnalyzer())
#-------------------
# Instantiate plugin
#-------------------
trackmate = TrackMate(model, settings)
#--------
# Process
#--------
ok = trackmate.checkInput()
if not ok:
sys.exit(str(trackmate.getErrorMessage()))
ok = trackmate.process()
if not ok:
sys.exit(str(trackmate.getErrorMessage()))
#----------------
# Display results
#----------------
selectionModel = SelectionModel(model)
displayer = HyperStackDisplayer(model, selectionModel, imp)
displayer.render()
displayer.refresh()
# Echo results with the logger we set at start:
#model.getLogger().log(str(model))
fm = model.getFeatureModel()
norm_x = []
norm_y = []
for id in model.getTrackModel().trackIDs(True):
track = model.getTrackModel().trackSpots(id)
for spot in track:
t = spot.getFeature('FRAME')
if (t == 0.0):
min_x = spot.getFeature('POSITION_X')
min_y = spot.getFeature('POSITION_Y')
for spot in track:
norm_x.append(spot.getFeature('POSITION_X') - min_x)
norm_y.append(spot.getFeature('POSITION_Y') - min_y)
max_x = abs(max(norm_x, key=abs))
max_y = abs(max(norm_y, key=abs))
return max_x, max_y
def track():
imp = IJ.getImage()
nChannels = imp.getNChannels() # Get the number of channels
orgtitle = imp.getTitle()
IJ.run("Subtract Background...", "rolling=50 sliding stack")
IJ.run("Enhance Contrast...", "saturated=0.3")
IJ.run("Multiply...", "value=10 stack")
IJ.run("Subtract Background...", "rolling=50 sliding stack")
IJ.run("Set Scale...", "distance=0")
channels = ChannelSplitter.split(imp)
imp_GFP = channels[0]
imp_RFP = channels[1]
IJ.selectWindow(orgtitle)
IJ.run("Close")
ic = ImageCalculator()
imp_merge = ic.run("Add create stack", imp_GFP, imp_RFP)
imp_merge.setTitle("add_channels")
imp_merge.show()
imp_RFP.show()
imp_GFP.show()
imp5 = ImagePlus()
IJ.run(imp5, "Merge Channels...", "c1=[" + imp_merge.title + "] c2=" + imp_GFP.title + ' c3=' + imp_RFP.title + " create")
print("c1=[" + imp_merge.title + "] c2=" + imp_GFP.title + ' c3=' + imp_RFP.title + " create")
imp5.show()
imp5 = IJ.getImage()
nChannels = imp5.getNChannels()
# Setup settings for TrackMate
settings = Settings()
settings.setFrom(imp5)
# Spot analyzer: we want the multi-C intensity analyzer.
settings.addSpotAnalyzerFactory(SpotMultiChannelIntensityAnalyzerFactory())
# Spot detector.
settings.detectorFactory = LogDetectorFactory()
settings.detectorSettings = settings.detectorFactory.getDefaultSettings()
settings.detectorSettings['TARGET_CHANNEL'] = 1
settings.detectorSettings['RADIUS'] = 24.0
settings.detectorSettings['THRESHOLD'] = 0.0
# Spot tracker.
# Configure tracker - We don't want to allow merges or splits
settings.trackerFactory = SparseLAPTrackerFactory()
settings.trackerSettings = LAPUtils.getDefaultLAPSettingsMap() # almost good enough
settings.trackerSettings['ALLOW_TRACK_SPLITTING'] = False
settings.trackerSettings['ALLOW_TRACK_MERGING'] = False
settings.trackerSettings['LINKING_MAX_DISTANCE'] = 8.0
settings.trackerSettings['GAP_CLOSING_MAX_DISTANCE'] = 8.0
settings.trackerSettings['MAX_FRAME_GAP'] = 1
# Configure track filters
settings.addTrackAnalyzer(TrackDurationAnalyzer())
settings.addTrackAnalyzer(TrackSpotQualityFeatureAnalyzer())
filter1 = FeatureFilter('TRACK_DURATION', 20, True)
settings.addTrackFilter(filter1)
# Run TrackMate and store data into Model.
model = Model()
trackmate = TrackMate(model, settings)
ok = trackmate.checkInput()
if not ok:
sys.exit(str(trackmate.getErrorMessage()))
ok = trackmate.process()
if not ok:
sys.exit(str(trackmate.getErrorMessage()))
selectionModel = SelectionModel(model)
displayer = HyperStackDisplayer(model, selectionModel, imp5)
displayer.render()
displayer.refresh()
IJ.log('TrackMate completed successfully.')
IJ.log('Found %d spots in %d tracks.' % (model.getSpots().getNSpots(True) , model.getTrackModel().nTracks(True)))
# Print results in the console.
headerStr = '%10s %10s %10s %10s %10s %10s' % ('Spot_ID', 'Track_ID', 'Frame', 'X', 'Y', 'Z')
rowStr = '%10d %10d %10d %10.1f %10.1f %10.1f'
for i in range( nChannels ):
headerStr += (' %10s' % ( 'C' + str(i+1) ) )
rowStr += ( ' %10.1f' )
#open a file to save results
myfile = open('/home/rickettsia/Desktop/data/Clamydial_Image_Analysis/EMS_BMECBMELVA_20X_01122019/data/'+orgtitle.split('.')[0]+'.csv', 'wb')
wr = csv.writer(myfile, quoting=csv.QUOTE_ALL)
wr.writerow(['Spot_ID', 'Track_ID', 'Frame', 'X', 'Y', 'Z', 'Channel_1', 'Channel_2'])
IJ.log('\n')
IJ.log(headerStr)
tm = model.getTrackModel()
trackIDs = tm.trackIDs(True)
for trackID in trackIDs:
spots = tm.trackSpots(trackID)
# Let's sort them by frame.
ls = ArrayList(spots)
for spot in ls:
values = [spot.ID(), trackID, spot.getFeature('FRAME'), \
spot.getFeature('POSITION_X'), spot.getFeature('POSITION_Y'), spot.getFeature('POSITION_Z')]
for i in range(nChannels):
values.append(spot.getFeature('MEAN_INTENSITY%02d' % (i+1)))
IJ.log(rowStr % tuple(values))
l1 = (values[0], values[1], values[2], values[3], values[4], values[5], values[7], values[8])
wr.writerow(l1)
myfile.close()
IJ.selectWindow("Merged")
IJ.run("Close")
import fiji.plugin.trackmate.action.ExportTracksToXML
# Swap Z and T dimensions if T=1
dims = imp.getDimensions() # default order: XYCZT
if (dims[4] == 1) {
imp.setDimensions( dims[2,4,3] )
}
# Setup settings for TrackMate
settings = new Settings()
settings.setFrom(imp)
settings.dt = 0.05
settings.detectorFactory = new LogDetectorFactory()
settings.detectorSettings = settings.detectorFactory.getDefaultSettings()
println settings.detectorSettings
settings.detectorSettings['RADIUS'] = radius
settings.detectorSettings['THRESHOLD'] = threshold
println settings.detectorSettings
settings.trackerFactory = new SparseLAPTrackerFactory()
settings.trackerSettings = LAPUtils.getDefaultLAPSettingsMap()
settings.trackerSettings['MAX_FRAME_GAP'] = frameGap
settings.trackerSettings['LINKING_MAX_DISTANCE'] = linkingMax
settings.trackerSettings['GAP_CLOSING_MAX_DISTANCE'] = closingMax
def run_trackmate(
imp, path_out="./", detector="log", radius=2.5, threshold=0.0, median_filter=False
):
"""Log Trackmate detection run with given parameters.
Saves spots in a csv file in the given path_out with encoded parameters.
Args:
imp: ImagePlus to be processed
path_out: Output directory to save files.
detector: Type of detection method. Options are 'log', 'dog'.
radius: Radius of spots in pixels.
threshold: Threshold value to filter spots.
median_filter: True if median_filtering should be used.
"""
if imp.dimensions[2] != 1:
raise ValueError(
"Imp's dimensions must be [n, n, 1] but are " + imp.dimensions[2]
)
# Create the model object now
model = Model()
model.setLogger(Logger.VOID_LOGGER)
# Prepare settings object
settings = Settings()
settings.setFrom(imp)
# Configure detector
settings.detectorFactory = (
DogDetectorFactory() if detector == "dog" else LogDetectorFactory()
)
settings.detectorSettings = {
"DO_SUBPIXEL_LOCALIZATION": True,
"RADIUS": radius,
"TARGET_CHANNEL": 1,
"THRESHOLD": threshold,
"DO_MEDIAN_FILTERING": median_filter,
}
# Instantiate plugin
trackmate = TrackMate(model, settings)
# Process
# output = trackmate.process()
output = trackmate.execDetection()
if not output:
print("error process")
return None
# Get output from a single image
fname = str(imp.title)
spots = [["fname", "detector", "radius", "threshold", "median", "x", "y", "q"]]
for spot in model.spots.iterator(0):
x = spot.getFeature("POSITION_X")
y = spot.getFeature("POSITION_Y")
q = spot.getFeature("QUALITY")
spots.append([fname, detector, radius, threshold, median_filter, x, y, q])
# Save output
outname = os.path.splitext(os.path.basename(fname))[0] + "_" + str(radius) + ".csv"
with open(os.path.join(path_out, outname), "wb") as f:
wr = csv.writer(f)
for row in spots:
wr.writerow(row)
def run_process(input_path, output_path):
IJ.run("Image Sequence...", "open=[" + input_path + "] convert sort")
imp = WindowManager.getCurrentImage()
dims = imp.getDimensions()
imp.setDimensions(dims[2], dims[4], dims[3])
dims = imp.getDimensions()
ImageConverter(imp).convertToGray8()
model = Model()
model.setLogger(Logger.IJ_LOGGER)
settings = Settings()
settings.setFrom(imp)
settings.detectorFactory = LogDetectorFactory()
settings.detectorSettings = {
'DO_SUBPIXEL_LOCALIZATION': True,
'RADIUS': float(11.0),
'THRESHOLD': float(0.0),
'DO_MEDIAN_FILTERING': True
}
# Configure spot filters - Classical filter on quality
# filter1 = FeatureFilter('QUALITY', 30, True)
# settings.addSpotFilter(filter1)
settings.trackerFactory = SparseLAPTrackerFactory()
settings.trackerSettings = LAPUtils.getDefaultLAPSettingsMap(
) #this sets tens of madatory settings
settings.trackerSettings['LINKING_MAX_DISTANCE'] = 15.0
settings.trackerSettings['GAP_CLOSING_MAX_DISTANCE'] = 15.0
settings.trackerSettings['MAX_FRAME_GAP'] = 2
# print(LAPUtils.getDefaultLAPSettingsMap())
#
# settings.trackerFactory = SimpleLAPTrackerFactory()
## settings.trackerSettings = LAPUtils.SimpleLAPTrackerFactory() #this sets tens of madatory settings
# settings.trackerSettings['LINKING_MAX_DISTANCE'] = 15.0
# settings.trackerSettings['GAP_CLOSING_MAX_DISTANCE']=15.0
# settings.trackerSettings['MAX_FRAME_GAP']= 2
# settings.trackerSettings['ALLOW_GAP_CLOSING']= True
# settings.trackerSettings['ALLOW_TRACK_SPLITTING']= False
# settings.trackerSettings['ALLOW_TRACK_MERGING']= False
# settings.trackerSettings['SPLITTING_MAX_DISTANCE']= 1000.0
# settings.trackerSettings['MERGING_MAX_DISTANCE']= 1000.0
# settings.trackerSettings['ALTERNATIVE_LINKING_COST_FACTOR']= 1000.0
# # ?
# settings.trackerSettings['CUTOFF_PERCENTILE']= 1000.0
# settings.trackerSettings['BLOCKING_VALUE']= 1000.0
#
# filter2 = FeatureFilter('NUMBER_OF_SPOTS_IN_TRACK', 6.86, True)
# settings.addTrackFilter(filter2)
trackmate = TrackMate(model, settings)
ok = trackmate.checkInput()
if not ok:
sys.exit(str(trackmate.getErrorMessage()))
raise Exception("trackmate: checkInput failed")
# if ok:
# print("Input ok")
ok = trackmate.process()
if not ok:
raise Exception("trackmate: process failed")
# if ok:
# print("Process ok")
#----------------
# Display results
#----------------
model.getLogger().log('Found ' + str(model.getTrackModel().nTracks(True)) +
' tracks.')
selectionModel = SelectionModel(model)
displayer = HyperStackDisplayer(model, selectionModel, imp)
# displayer.render()
# The feature model, that stores edge and track features.
fm = model.getFeatureModel()
# print(fm)
labels_row = [
'id', 'spot_id', 'x', 'y', 'frame', 'quality', 'type', 'length'
]
#'qualitiy','visability', 'track_length']
#rows = []
track_ids = model.getTrackModel().trackIDs(True)
with open(output_path, 'w') as file:
writer = csv.writer(file)
writer.writerow(labels_row)
for id in track_ids:
# Fetch the track feature from the feature model.
# model.getLogger().log('')
# model.getLogger().log('Track ' + str(id) + ': mean velocity = ' + str(v) + ' ' + model.getSpaceUnits() + '/' + model.getTimeUnits())
track = model.getTrackModel().trackSpots(id)
num_spots = track.size()
for spot in track:
# print(spot.getFeatures())
row = []
row.append(id)
sid = spot.ID()
row.append(sid)
# Fetch spot features directly from spot.
x = spot.getFeature('POSITION_X')
row.append(x)
y = spot.getFeature('POSITION_Y')
row.append(y)
t = spot.getFeature('FRAME')
row.append(int(t))
# print("x: {} y: {} t: {}".format(x, y, t))
q = spot.getFeature('QUALITY')
row.append(q)
# snr=spot.getFeature('SNR')
# row.append(snr)
# mean=spot.getFeature('MEAN_INTENSITY')
# row.append(mean)
# visibility=spot.getFeature('VISIBILITY')
# print(visibility)
# break
# row.append(visability)
# model.getLogger().log('\tspot ID = ' + str(sid) + ': x='+str(x)+', y='+str(y)+', t='+str(t)+', q='+str(q) + ', snr='+str(snr) + ', mean = ' + str(mean))
row.append(1)
row.append(num_spots)
writer.writerow(row)
# rows.append(row)
file.close()
return IJ
def track_cells(folder_w, filename, imp, correction):
#imp = IJ.openImage(os.path.join(folder,filename))
#imp.show()
#get image dimensions, set ROI remove part of flouresncent ring
x_size = ImagePlus.getDimensions(imp)[0]
y_size = ImagePlus.getDimensions(imp)[1]
x_start = 0
y_start = 0
#calculate alternative ROI
if crop_ring:
x_start = 170 / 2
y_start = 170 / 2
x_size = x_size - 170
y_size = y_size - 170
print(
str(x_start) + ", " + str(y_start) + ", " + str(x_size) + ", " +
str(y_size))
imp.setRoi(OvalRoi(x_start, y_start, x_size, y_size))
#imp_dup = imp.duplicate()
#imp_dup.show()
#red_corrected_img.show()
IJ.run(imp, "Make Inverse", "")
IJ.setForegroundColor(0, 0, 0)
IJ.run(imp, "Fill", "stack")
imp.killRoi()
#imp.show()
#sys.exit()
#img_filename = filename+"_corrected_red_stack.tif"
#folder_filename= os.path.join(well_folder,img_filename)
#IJ.save(imp, folder_filename)
#----------------------------
# Create the model object now
#----------------------------
# Some of the parameters we configure below need to have
# a reference to the model at creation. So we create an
# empty model now.
model = Model()
# Send all messages to ImageJ log window.
model.setLogger(Logger.IJ_LOGGER)
#------------------------
# Prepare settings object
#------------------------
settings = Settings()
settings.setFrom(imp)
# Configure detector - We use the Strings for the keys
settings.detectorFactory = LogDetectorFactory()
settings.detectorSettings = {
'DO_SUBPIXEL_LOCALIZATION': SUBPIXEL_LOCALIZATION,
'RADIUS': RADIUS,
'TARGET_CHANNEL': TARGET_CHANNEL,
'THRESHOLD': THRESHOLD,
'DO_MEDIAN_FILTERING': MEDIAN_FILTERING,
}
# Configure spot filters - Classical filter on quality
settings.initialSpotFilterValue = SPOT_FILTER
settings.addSpotAnalyzerFactory(SpotIntensityAnalyzerFactory())
settings.addSpotAnalyzerFactory(SpotContrastAndSNRAnalyzerFactory())
settings.addSpotAnalyzerFactory(SpotMorphologyAnalyzerFactory())
settings.addSpotAnalyzerFactory(SpotRadiusEstimatorFactory())
filter1 = FeatureFilter('QUALITY', QUALITY, True)
filter2 = FeatureFilter('CONTRAST', CONTRAST, True)
filter2a = FeatureFilter('ESTIMATED_DIAMETER', MAX_ESTIMATED_DIAMETER,
False)
filter2b = FeatureFilter('MEDIAN_INTENSITY', MAX_MEDIAN_INTENSITY, False)
settings.addSpotFilter(filter1)
settings.addSpotFilter(filter2)
settings.addSpotFilter(filter2a)
settings.addSpotFilter(filter2b)
print(settings.spotFilters)
# Configure tracker - We want to allow merges and fusions
settings.trackerFactory = SparseLAPTrackerFactory()
settings.trackerSettings = LAPUtils.getDefaultLAPSettingsMap(
) # almost good enough
##adapted from https://forum.image.sc/t/trackmate-scripting-automatically-exporting-spots-in-tracks-links-in-tracks-tracks-statistics-and-branching-analysis-to-csv/6256
#linking settings
settings.trackerSettings['LINKING_MAX_DISTANCE'] = LINKING_MAX_DISTANCE
if LINKING_FEATURE_PENALTIES == True:
settings.trackerSettings['LINKING_FEATURE_PENALTIES'] = {
LINKING_FEATURE_PENALTIES_TYPE: LINKING_FEATURE_PENALTIES_VALUE
}
else:
settings.trackerSettings['LINKING_FEATURE_PENALTIES'] = {}
#gap closing settings
settings.trackerSettings['ALLOW_GAP_CLOSING'] = ALLOW_GAP_CLOSING
if ALLOW_GAP_CLOSING == True:
settings.trackerSettings[
'GAP_CLOSING_MAX_DISTANCE'] = GAP_CLOSING_MAX_DISTANCE
settings.trackerSettings['MAX_FRAME_GAP'] = MAX_FRAME_GAP
if GAP_CLOSING_FEATURE_PENALTIES == True:
settings.trackerSettings['GAP_CLOSING_FEATURE_PENALTIES'] = {
GAP_CLOSING_FEATURE_PENALTIES_TYPE:
GAP_CLOSING_FEATURE_PENALTIES_VALUE
}
else:
settings.trackerSettings['GAP_CLOSING_FEATURE_PENALTIES'] = {}
#splitting settings
settings.trackerSettings['ALLOW_TRACK_SPLITTING'] = ALLOW_TRACK_SPLITTING
if ALLOW_TRACK_SPLITTING == True:
settings.trackerSettings[
'SPLITTING_MAX_DISTANCE'] = SPLITTING_MAX_DISTANCE
if SPLITTING_FEATURE_PENALTIES == True:
settings.trackerSettings['SPLITTING_FEATURE_PENALTIES'] = {
SPLITTING_FEATURE_PENALTIES_TYPE:
SPLITTING_FEATURE_PENALTIES_VALUE
}
else:
settings.trackerSettings['SPLITTING_FEATURE_PENALTIES'] = {}
#merging settings
settings.trackerSettings['ALLOW_TRACK_MERGING'] = ALLOW_TRACK_MERGING
if ALLOW_TRACK_MERGING == True:
settings.trackerSettings['MERGING_MAX_DISTANCE'] = MERGING_MAX_DISTANCE
if MERGING_FEATURE_PENALTIES == True:
settings.trackerSettings['MERGING_FEATURE_PENALTIES'] = {
MERGING_FEATURE_PENALTIES_TYPE: MERGING_FEATURE_PENALTIES_VALUE
}
else:
settings.trackerSettings['MERGING_FEATURE_PENALTIES'] = {}
print(settings.trackerSettings)
# Configure track analyzers - Later on we want to filter out tracks
# based on their displacement, so we need to state that we want
# track displacement to be calculated. By default, out of the GUI,
# not features are calculated.
# The displacement feature is provided by the TrackDurationAnalyzer.
settings.addTrackAnalyzer(TrackDurationAnalyzer())
settings.addTrackAnalyzer(TrackSpotQualityFeatureAnalyzer())
# Configure track filters - We want to get rid of the two immobile spots at
# the bottom right of the image. Track displacement must be above 10 pixels.
filter3 = FeatureFilter('TRACK_DISPLACEMENT', TRACK_DISPLACEMENT, True)
filter4 = FeatureFilter('TRACK_START', TRACK_START, False)
#filter5 = FeatureFilter('TRACK_STOP', float(imp.getStack().getSize())-1.1, True)
settings.addTrackFilter(filter3)
settings.addTrackFilter(filter4)
#settings.addTrackFilter(filter5)
#-------------------
# Instantiate plugin
#-------------------
trackmate = TrackMate(model, settings)
#--------
# Process
#--------
ok = trackmate.checkInput()
if not ok:
sys.exit(str(trackmate.getErrorMessage()))
ok = trackmate.process()
# if not ok:
#sys.exit(str(trackmate.getErrorMessage()))
#----------------
# Display results
#----------------
#Set output folder and filename and create output folder
well_folder = os.path.join(folder_w, filename)
output_folder = os.path.join(well_folder, "Tracking")
create_folder(output_folder)
xml_file_name = filename + "_" + correction + "_trackmate_analysis.xml"
folder_filename_xml = os.path.join(output_folder, xml_file_name)
#ExportTracksToXML.export(model, settings, File(folder_filename_xml))
outfile = TmXmlWriter(File(folder_filename_xml))
outfile.appendSettings(settings)
outfile.appendModel(model)
outfile.writeToFile()
# Echo results with the logger we set at start:
#model.getLogger().log(str(model))
#create araray of timepoint length with filled 0
cell_counts = zerolistmaker(imp.getStack().getSize())
if ok:
for id in model.getTrackModel().trackIDs(True):
# Fetch the track feature from the feature model.
track = model.getTrackModel().trackSpots(id)
for spot in track:
# Fetch spot features directly from spot.
t = spot.getFeature('FRAME')
print(t)
cell_counts[int(t)] = cell_counts[int(t)] + 1
else:
print("No spots detected!")
if HEADLESS == False:
selectionModel = SelectionModel(model)
displayer = HyperStackDisplayer(model, selectionModel, imp)
displayer.render()
displayer.refresh()
del imp
return (cell_counts + [len(model.getTrackModel().trackIDs(True))])
def run_trackmate(imp, path, filename, params, batch_mode=False):
# initialize trackmate model
model = Model()
# Set logger - use to see outputs, not needed in batch mode
model.setLogger(Logger.IJ_LOGGER)
# Create setting object from image
settings = Settings()
settings.setFrom(imp)
cal = imp.getCalibration()
model.setPhysicalUnits("micron", "sec")
# Configure detector
settings.detectorFactory = LogDetectorFactory()
# settings.detectorFactory = DogDetectorFactory()
settings.detectorSettings = {
'DO_SUBPIXEL_LOCALIZATION': params.do_subpixel_localization,
'RADIUS': params.radius,
'TARGET_CHANNEL': 0,
'THRESHOLD': params.threshold,
'DO_MEDIAN_FILTERING': params.do_median_filtering,
}
# print(params)
# Add spot filters
filter_quality = FeatureFilter('QUALITY', params.quality, True)
settings.addSpotFilter(filter_quality)
filter_snr = FeatureFilter('SNR', params.snr, True)
settings.addSpotFilter(filter_snr)
# Compute spot features
settings.addSpotAnalyzerFactory(SpotIntensityAnalyzerFactory())
settings.addSpotAnalyzerFactory(SpotContrastAndSNRAnalyzerFactory())
# Compute track features
settings.addTrackAnalyzer(TrackBranchingAnalyzer())
settings.addTrackAnalyzer(TrackDurationAnalyzer())
settings.addTrackAnalyzer(TrackIndexAnalyzer())
settings.addTrackAnalyzer(TrackLocationAnalyzer())
settings.addTrackAnalyzer(TrackSpeedStatisticsAnalyzer())
settings.addTrackAnalyzer(TrackSpotQualityFeatureAnalyzer())
# Update model
ModelFeatureUpdater(model, settings)
# Configure tracker
settings.trackerFactory = SparseLAPTrackerFactory()
settings.trackerSettings = LAPUtils.getDefaultLAPSettingsMap()
settings.trackerSettings[
'LINKING_MAX_DISTANCE'] = params.linking_max_distance
settings.trackerSettings[
'GAP_CLOSING_MAX_DISTANCE'] = params.gap_closing_max_distance
settings.trackerSettings['MAX_FRAME_GAP'] = params.max_frame_gap
# Add track filters
filter_T1 = FeatureFilter('TRACK_DURATION', params.track_duration, True)
filter_MTD = FeatureFilter('TRACK_DISPLACEMENT', params.track_displacement,
True)
settings.addTrackFilter(filter_T1)
settings.addTrackFilter(filter_MTD)
# Instantiate trackmate
trackmate = TrackMate(model, settings)
# Execute all
ok = trackmate.checkInput()
if not ok:
IJ.showMessage("No spots found... Adjust detection parameter.\n" +
str(trackmate.getErrorMessage()))
sys.exit(str(trackmate.getErrorMessage()))
ok = trackmate.process()
if not ok:
IJ.showMessage("No spots found... Adjust detection parameter.\n" +
str(trackmate.getErrorMessage()))
sys.exit(str(trackmate.getErrorMessage()))
filename = os.path.splitext(filename)[0] #filename without extension
outFile = File(os.path.join(path, filename + "_Tracks.xml"))
ExportTracksToXML.export(model, settings, outFile)
#imp.close()
tm_writer = TmXmlWriter(File(os.path.join(path, filename + "_TM.xml")))
tm_writer.appendModel(model)
tm_writer.appendSettings(settings)
tm_writer.writeToFile()
if not batch_mode:
selectionModel = SelectionModel(model)
displayer = HyperStackDisplayer(model, selectionModel, imp)
displayer.render()
displayer.refresh()
# Echo results with the logger we set at start:
model.getLogger().log(str(model))
def getSpots(imp,
channel,
detector_type,
radius,
threshold,
overlay,
roi_type="large",
roi_color=ColorRGB("blue")):
""" Performs the detection, adding spots to the image overlay
:imp: The image (ImagePlus) being analyzed
:channel: The target channel
:detector_type: A string describing the detector: "LoG" or "DoG"
:radius: Spot radius (NB: trackmate GUI accepts diameter)
:threshold: Quality cutoff value
:overlay: The image overlay to store spot (MultiPoint) ROIs
:roi_type: A string describing how spot ROIs should be displayed
:returns: The n. of detected spots
"""
settings = Settings()
settings.setFrom(imp)
settings.detectorFactory = (LogDetectorFactory() if "LoG" in detector_type
else DogDetectorFactory())
settings.detectorSettings = {
DK.KEY_DO_SUBPIXEL_LOCALIZATION: False,
DK.KEY_DO_MEDIAN_FILTERING: True,
DK.KEY_TARGET_CHANNEL: channel,
DK.KEY_RADIUS: radius,
DK.KEY_THRESHOLD: threshold,
}
trackmate = TrackMate(settings)
if not trackmate.execDetection():
lservice.error(str(trackmate.getErrorMessage()))
return 0
model = trackmate.model
spots = model.getSpots()
count = spots.getNSpots(False)
ch_id = "Spots Ch%d" % channel
if count > 0:
roi = None
cal = imp.getCalibration()
t_pos = imp.getT()
if (t_pos > 1):
lservice.warn("Only frame %d was considered..." % t_pos)
for spot in spots.iterable(False):
x = cal.getRawX(spot.getFeature(spot.POSITION_X))
y = cal.getRawY(spot.getFeature(spot.POSITION_Y))
z = spot.getFeature(spot.POSITION_Z)
if z == 0 or not cal.pixelDepth or cal.pixelDepth == 0:
z = 1
else:
z = int(z // cal.pixelDepth)
imp.setPosition(channel, z, t_pos)
if roi is None:
roi = PointRoi(int(x), int(y), imp)
else:
roi.addPoint(imp, x, y)
roi.setStrokeColor(colorRGBtoColor(roi_color))
if "large" in roi_type:
roi.setPointType(3)
roi.setSize(4)
else:
roi.setPointType(2)
roi.setSize(1)
overlay.add(roi, ch_id)
return count