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"""Tools to perform particle tracking with Trackmate ImageJ plugin. |
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This module includes functions for prepping images and performing tracking |
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using the TrackMate ImageJ plugin [1]_. |
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References |
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---------- |
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.. [1] Tinevez, JY,; Perry, N. & Schindelin, J. et al. (2016), "TrackMate: an |
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open and extensible platoform for single-particle tracking.", Methods 115: |
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80-90, PMID 27713081 (on Google Scholar). |
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""" |
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import os |
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import sys |
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import subprocess |
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import tempfile |
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import random |
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from itertools import compress |
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import os.path as op |
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import numpy as np |
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import skimage.io as sio |
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import diff_classifier as dc |
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import diff_classifier.aws as aws |
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from sklearn import linear_model |
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from sklearn import svm |
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View Code Duplication |
def _get_fiji(): |
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"""Checks if Fiji is downloaded. |
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Checks if an existing version of Fiji is downloaded and whether |
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it is identified as the system variable "FIJI_BIN." Only compatible |
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with Linux and Mac systems. |
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""" |
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home = os.path.expanduser("~") |
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paths = [ |
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os.path.join(home, 'Applications/Fiji.app/Contents/MacOS/ImageJ-macosx'), |
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os.path.join(home, 'Fiji.app/Contents/MacOS/ImageJ-macosx'), |
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os.path.join(home, 'Fiji.app/ImageJ-linux64') |
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] |
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exists = [os.path.exists(p) for p in paths] |
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# paths = list(compress(paths, exists)) |
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# Currently only works on Mac and Linux |
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if sys.platform != 'darwin' and not sys.platform.startswith('linux'): |
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# print('System is not Linux or Mac') |
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raise ValueError('System is not Linux or Mac') |
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# Has the user specified Fiji for us already as an environment variable? |
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# Want to use it if it is already installed |
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elif "FIJI_BIN" in os.environ: |
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print("FIJI_BIN defined.") |
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return os.environ["FIJI_BIN"] |
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# Is Fiji installed somewhere, but not defined in path? |
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# Checks in two locations, doesn't search entire system |
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elif any(exists): |
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print('Exists elsewhere') |
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counter = 0 |
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for exist in exists: |
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if exist: |
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path = paths[counter] |
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counter += 1 |
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return path |
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else: |
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# Download it if not |
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if sys.platform == 'darwin': |
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subprocess.run(['wget', 'https://downloads.imagej.net/fiji/latest/fiji-macosx.zip'], cwd=home) |
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subprocess.run(['unzip', 'fiji-macosx.zip'], cwd=home) |
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os.environ['FIJI_BIN'] = os.path.join(home, 'Fiji.app/Contents/MacOS/ImageJ-macosx') |
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elif sys.platform.startswith('linux'): |
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subprocess.run(['wget', 'https://downloads.imagej.net/fiji/latest/fiji-linux64.zip'], cwd=home) |
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subprocess.run(['unzip', 'fiji-linux64.zip'], cwd=home) |
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os.environ['FIJI_BIN'] = os.path.join(home, 'Fiji.app/ImageJ-linux64') |
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print("Downloaded Fiji") |
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return _get_fiji() |
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View Code Duplication |
def partition_im(tiffname, irows=4, icols=4, ores=(2048, 2048), |
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ires=(512, 512)): |
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"""Partitions image into smaller images. |
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Partitions a large image into irows x icols images of size ires and write |
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them to the drive. Default input image sizes are from a Nikon/Hamamatsu |
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camera setup (2048 x 2044 pixels). |
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Parameters |
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---------- |
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tiffname : string |
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Location of input image to be partitioned. |
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irows : int |
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Number of rows of size ires pixels to be partitioned from source image. |
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icols : int |
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Number of columns of size ires pixels to be partitioned from source image. |
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ores : tuple of int |
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Input images are scaled to size ores pixels prior to splitting. |
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ires : tuple of int |
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Output images are of size ires pixels. |
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Returns |
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-------- |
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names : list of str |
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List of output image filenames |
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Examples |
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-------- |
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>>> partition_im('your/sample/image.tif', irows=8, icols=8, ires=(256, 256)) |
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""" |
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test = sio.imread(tiffname) |
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oshape = test.shape |
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test2 = np.zeros((oshape[0], ores[0], ores[1]), dtype=test.dtype) |
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test2[0:oshape[0], 0:oshape[1], :] = test |
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new_image = np.zeros((oshape[0], ires[0], ires[1]), dtype=test.dtype) |
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names = [] |
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for row in range(irows): |
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for col in range(icols): |
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new_image = test2[:, row*ires[0]:(row+1)*ires[0], |
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col*ires[1]:(col+1)*ires[1]] |
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current = tiffname.split('.tif')[0] + '_%s_%s.tif' % (row, col) |
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sio.imsave(current, new_image) |
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names.append(current) |
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return names |
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def mean_intensity(local_im, frame=0): |
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"""Calculates mean intensity of first frame of input image. |
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Parameters |
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---------- |
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local_im : string |
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Location of input image. |
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frame : int |
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Frame at which to perform mean. |
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Returns |
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------- |
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test_intensity : float |
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Mean intensity of input image. |
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Examples |
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-------- |
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>>> mean_intensity('your/sample/image') |
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""" |
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test_image = sio.imread(local_im) |
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test_intensity = np.mean(test_image[frame, :, :]) |
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return test_intensity |
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View Code Duplication |
def track(target, out_file, template=None, fiji_bin=None, |
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tparams=None): |
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"""Performs particle tracking on input video. |
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Particle tracking is performed with the ImageJ plugin Trackmate. Outputs |
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a csv file containing analysis settings and particle trajectories. |
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Parameters |
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---------- |
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target : str |
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Full path to a tif file to do tracking on. Can also be a URL |
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(e.g., 'http://fiji.sc/samples/FakeTracks.tif') |
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out_file : str |
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Full path to a csv file to store the results. |
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template : str, optional |
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The full path of a template for tracking. Defaults to use |
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`data/trackmate_template.py` stored in the diff_classifier source-code. |
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fiji_bin : str |
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The full path to ImageJ executable file. Includes default search |
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locations for Mac and Linux systems. |
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radius : float |
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Estimated radius of particles in image. |
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threshold : float |
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Threshold value for particle detection step. |
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do_median_filtering : bool |
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If True, performs a median filter on video prior to tracking. |
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quality : float |
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Lower quality cutoff value for particle filtering. |
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xdims : tuple of int |
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Upper and lower x limits for particle filtering. |
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ydims : tuple of int |
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Upper and lower y limits for particle filtering. |
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median_intensity : float |
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Lower median intensity cutoff value for particle filtering. |
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snr : float |
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Lower signal to noise ratio cutoff value for particle filtering. |
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limking_max_distance : float |
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Maximum allowable distance in pixels between two frames to join |
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particles in track. |
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gap_closing_max_distance : float |
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Maximum allowable distance in pixels between more than two frames to |
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join particles in track. |
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max_frame_gap : int |
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Maximum allowable number of frames a particle is allowed to leave video |
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and be counted as same trajectory. |
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track_duration : float |
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Lower duration cutoff in frames for trajectory filtering. |
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""" |
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tdefault = {'frames': 651, 'radius': 3.0, 'threshold': 0.0, |
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'do_median_filtering': False, 'quality': 15.0, |
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'xdims': (0, 511), 'ydims': (1, 511), |
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'median_intensity': 300.0, |
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'snr': 0.0, 'linking_max_distance': 6.0, |
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'gap_closing_max_distance': 10.0, |
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'max_frame_gap': 3, 'track_duration': 20.0, |
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'detector': 'Dog'} |
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if tparams is None: |
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tparams = {} |
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for key, value in tdefault.items(): |
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if key not in tparams: |
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tparams[key] = value |
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if template is None: |
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template = op.join(op.split(dc.__file__)[0], |
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'data', |
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'trackmate_template3.py') |
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fiji_bin = _get_fiji() |
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script = ''.join(open(template).readlines()) |
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tpfile = tempfile.NamedTemporaryFile(suffix=".py") |
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fid = open(tpfile.name, 'w') |
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fid.write(script.format(target_file=target, frames=str(tparams['frames']), |
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radius=str(tparams['radius']), |
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threshold=str(tparams['threshold']), |
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do_median_filtering=str(tparams['do_median_filtering']), |
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quality=str(tparams['quality']), |
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xd=str(tparams['xdims'][1]), |
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yd=str(tparams['ydims'][1]), |
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ylo=str(tparams['ydims'][0]), |
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median_intensity=str(tparams['median_intensity']), |
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snr=str(tparams['snr']), |
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linking_max_distance=str(tparams['linking_max_distance']), |
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gap_closing_max_distance=str(tparams['gap_closing_max_distance']), |
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max_frame_gap=str(tparams['max_frame_gap']), |
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track_duration=str(tparams['track_duration']), |
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set_detector=str(tparams['detector']))) |
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fid.close() |
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cmd = "%s --ij2 --headless --run %s" % (fiji_bin, tpfile.name) |
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print(cmd) |
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subp = subprocess.run(cmd, stdout=subprocess.PIPE, shell=True) |
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fid = open(out_file, 'w') |
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fid.write(subp.stdout.decode()) |
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#print(subp.stdout.decode()) |
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fid.close() |
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View Code Duplication |
def regress_sys(folder, all_videos, yfit, training_size, randselect=True, |
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trainingdata=[], frame=0, have_output=True, download=True, |
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bucket_name='ccurtis.data'): |
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"""Uses regression based on image intensities to select tracking parameters. |
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This function uses regression methods from the scikit-learn module to |
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predict the lower quality cutoff values for particle filtering in TrackMate |
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based on the intensity distributions of input images. Currently only uses |
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the first frame of videos for analysis, and is limited to predicting |
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quality values. |
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In practice, users will run regress_sys twice in different modes to build |
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a regression system. First, set have_output to False. Function will return |
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list of randomly selected videos to include in the training dataset. The |
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user should then manually track particles using the Trackmate GUI, and enter |
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these values in during the next round as the input yfit variable. |
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Parameters |
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---------- |
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folder : str |
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S3 directory containing video files specified in all_videos. |
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all_videos: list of str |
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Contains prefixes of video filenames of entire video set to be |
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tracked. Training dataset will be some subset of these videos. |
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yfit: numpy.ndarray |
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Contains manually acquired quality levels using Trackmate for the |
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files contained in the training dataset. |
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training_size : int |
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Number of files in training dataset. |
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randselect : bool |
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If True, will randomly select training videos from all_videos. |
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If False, will use trainingdata as input training dataset. |
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trainingdata : list of str |
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Optional manually selected prefixes of video filenames to be |
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used as training dataset. |
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have_output: bool |
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If you have already acquired the quality values (yfit) for the |
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training dataset, set to True. If False, it will output the files |
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the user will need to acquire quality values for. |
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bucket_name : str |
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305
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|
|
S3 bucket containing videos to be downloaded for regression |
|
306
|
|
|
calculations. |
|
307
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|
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|
|
308
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Returns |
|
309
|
|
|
------- |
|
310
|
|
|
regress_object : list of sklearn.svm.classes. |
|
311
|
|
|
Contains list of regression objects assembled from the training |
|
312
|
|
|
datasets. Uses the mean, 10th percentile, 90th percentile, and |
|
313
|
|
|
standard deviation intensities to predict the quality parameter |
|
314
|
|
|
in Trackmate. |
|
315
|
|
|
tprefix : list of str |
|
316
|
|
|
Contains randomly selected images from all_videos to be included in |
|
317
|
|
|
training dataset. |
|
318
|
|
|
|
|
319
|
|
|
""" |
|
320
|
|
|
|
|
321
|
|
|
if randselect: |
|
322
|
|
|
tprefix = [] |
|
323
|
|
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for i in range(0, training_size): |
|
|
|
|
|
|
324
|
|
|
random.seed(i+1) |
|
325
|
|
|
tprefix.append(all_videos[random.randint(0, len(all_videos))]) |
|
326
|
|
|
if have_output is False: |
|
327
|
|
|
print("Get parameters for: {}".format(tprefix[i])) |
|
328
|
|
|
else: |
|
329
|
|
|
tprefix = trainingdata |
|
330
|
|
|
|
|
331
|
|
|
if have_output is True: |
|
332
|
|
|
# Define descriptors |
|
333
|
|
|
descriptors = np.zeros((training_size, 4)) |
|
334
|
|
|
counter = 0 |
|
335
|
|
|
for name in tprefix: |
|
336
|
|
|
local_im = name + '.tif' |
|
337
|
|
|
remote_im = "{}/{}".format(folder, local_im) |
|
338
|
|
|
if download: |
|
339
|
|
|
aws.download_s3(remote_im, local_im, bucket_name=bucket_name) |
|
340
|
|
|
test_image = sio.imread(local_im) |
|
341
|
|
|
descriptors[counter, 0] = np.mean(test_image[frame, :, :]) |
|
342
|
|
|
descriptors[counter, 1] = np.std(test_image[frame, :, :]) |
|
343
|
|
|
descriptors[counter, 2] = np.percentile(test_image[frame, :, :], 10) |
|
344
|
|
|
descriptors[counter, 3] = np.percentile(test_image[frame, :, :], 90) |
|
345
|
|
|
counter = counter + 1 |
|
346
|
|
|
|
|
347
|
|
|
# Define regression techniques |
|
348
|
|
|
xfit = descriptors |
|
349
|
|
|
classifiers = [ |
|
350
|
|
|
svm.SVR(), |
|
351
|
|
|
linear_model.SGDRegressor(), |
|
352
|
|
|
linear_model.BayesianRidge(), |
|
353
|
|
|
linear_model.LassoLars(), |
|
354
|
|
|
linear_model.ARDRegression(), |
|
355
|
|
|
linear_model.PassiveAggressiveRegressor(), |
|
356
|
|
|
linear_model.TheilSenRegressor(), |
|
357
|
|
|
linear_model.LinearRegression()] |
|
358
|
|
|
|
|
359
|
|
|
regress_object = [] |
|
360
|
|
|
for item in classifiers: |
|
361
|
|
|
clf = item |
|
362
|
|
|
regress_object.append(clf.fit(xfit, yfit)) |
|
363
|
|
|
|
|
364
|
|
|
return regress_object |
|
365
|
|
|
|
|
366
|
|
|
else: |
|
367
|
|
|
return tprefix |
|
368
|
|
|
|
|
369
|
|
|
|
|
370
|
|
View Code Duplication |
def regress_tracking_params(regress_object, to_track, |
|
|
|
|
|
|
371
|
|
|
regmethod='LinearRegression', frame=0): |
|
372
|
|
|
"""Predicts quality values to be used in particle tracking. |
|
373
|
|
|
|
|
374
|
|
|
Uses the regress object from regress_sys to predict tracking |
|
375
|
|
|
parameters for TrackMate analysis. |
|
376
|
|
|
|
|
377
|
|
|
Parameters |
|
378
|
|
|
---------- |
|
379
|
|
|
regress_object: list of sklearn.svm.classes. |
|
380
|
|
|
Obtained from regress_sys |
|
381
|
|
|
to_track: string |
|
382
|
|
|
Prefix of video files to be tracked. |
|
383
|
|
|
regmethod: {'LinearRegression', 'SVR', 'SGDRegressor', 'BayesianRidge', |
|
384
|
|
|
'LassoLars', 'ARDRegression', 'PassiveAggressiveRegressor', |
|
385
|
|
|
'TheilSenRegressor'} |
|
386
|
|
|
Desired regression method. |
|
387
|
|
|
|
|
388
|
|
|
Returns |
|
389
|
|
|
------- |
|
390
|
|
|
fqual: float |
|
391
|
|
|
Predicted quality factor used in TrackMate analysis. |
|
392
|
|
|
|
|
393
|
|
|
""" |
|
394
|
|
|
|
|
395
|
|
|
local_im = to_track + '.tif' |
|
396
|
|
|
pX = np.zeros((1, 4)) |
|
397
|
|
|
test_image = sio.imread(local_im) |
|
398
|
|
|
pX[0, 0] = np.mean(test_image[frame, :, :]) |
|
399
|
|
|
pX[0, 1] = np.std(test_image[frame, :, :]) |
|
400
|
|
|
pX[0, 2] = np.percentile(test_image[frame, :, :], 10) |
|
401
|
|
|
pX[0, 3] = np.percentile(test_image[frame:, :, :], 90) |
|
402
|
|
|
|
|
403
|
|
|
quality = [] |
|
404
|
|
|
for item in regress_object: |
|
405
|
|
|
quality.append(item.predict(pX)[0]) |
|
406
|
|
|
|
|
407
|
|
|
if regmethod == 'SVR': |
|
408
|
|
|
fqual = quality[0] |
|
409
|
|
|
elif regmethod == 'SGDRegressor': |
|
410
|
|
|
fqual = quality[1] |
|
411
|
|
|
elif regmethod == 'BayesianRidge': |
|
412
|
|
|
fqual = quality[2] |
|
413
|
|
|
elif regmethod == 'LassoLars': |
|
414
|
|
|
fqual = quality[3] |
|
415
|
|
|
elif regmethod == 'ARDRegression': |
|
416
|
|
|
fqual = quality[4] |
|
417
|
|
|
elif regmethod == 'PassiveAggressiveRegressor': |
|
418
|
|
|
fqual = quality[5] |
|
419
|
|
|
elif regmethod == 'TheilSenRegressor': |
|
420
|
|
|
fqual = quality[6] |
|
421
|
|
|
elif regmethod == 'LinearRegression': |
|
422
|
|
|
fqual = quality[7] |
|
423
|
|
|
else: |
|
424
|
|
|
fqual = 3.0 |
|
425
|
|
|
|
|
426
|
|
|
return fqual |
|
427
|
|
|
|