lipase/preprocessing.py

347 lines
14 KiB
Python

"""preprocessing of synchrotron images based on telemosToolbox."""
from ij import IJ
from ij.plugin import ImageCalculator
from catalog import ImageCatalog, Sequence
def imagej_run_image_command(image, command, options):
"""performs the given imagej command on the given image
:param ImagePlus image:
:param str command: imagej command (eg "Gray Morphology")
:param str options: imagej options (eg "radius=1 type=square operator=open")
wrapper around IJ.run (https://imagej.nih.gov/ij/developer/api/ij/IJ.html#run-ij.ImagePlus-java.lang.String-java.lang.String-) which raises an exception on error
"""
IJ.run(image, command, options)
error_message = IJ.getErrorMessage()
if error_message is not None:
raise Exception('The command "%s" with options "%s" failed because of the following error : %s' % (command, options, error_message))
def compute_max(images_file_path):
"""Computes for each pixel position the maximum at this position in all input images.
:param list(str) images_file_path:
:rtype ImagePlus:
"""
assert len(images_file_path) > 1
max_image = IJ.openImage(images_file_path[0])
print('max_image', max_image)
for image_file_path in images_file_path[2:-1]:
other_image = IJ.openImage(image_file_path)
print('other_image', other_image)
ic = ImageCalculator()
ic.run("max", max_image, other_image)
print('max_image', max_image)
return max_image
def replace_border(image, band_width):
"""Overwrites the outer band of the image by duplicating the value of the pixels that touch this outer band
:param ImagePlus image:
:param int band_width: width of the outer band, in pixels
:rtype ImagePlus:
adaptation for the following code from matlab telemosToolbx's estimatedwhiteFluoImageTelemos function
outer = white_estimate;
white_estimate = white_estimate(4:(end-3),4:(end-3));
outer(1:3,4:(end-3))=repmat(white_estimate(1,:),3,1); # top
outer((end-2):end,4:(end-3))=repmat(white_estimate(end,:),3,1); # bottom
outer(:,1:3)=repmat(outer(:,4),1,3); # left
outer(:,(end-2):end)=repmat(outer(:,end-3),1,3); # right
white_estimate = outer;
clear outer;
"""
raise NotImplementedError()
return image
def perform_gray_morphology(image, operator, structuring_element_shape, structuring_element_radius):
"""
:param ImagePlus image:
:param str operator: eg 'open'
:param str structuring_element_shape: eg 'square'
:param int structuring_element_radius:
"""
assert operator not in ['fast open', 'fast erode'], "as of 13/09/2019, fast operators such as 'fast erode' seem broken in fiji (the resulting image is not at all similar to their slow equivalent)"
processor = image.getProcessor()
convert_to_byte = False
if processor.getBitDepth() != 8:
convert_to_byte = True
min_value = processor.getMin()
max_value = processor.getMax()
print("warning: downgrading image to byte as imagej's Gray Morphology processing doesn't handle 16bit images (range=[%d, %d])" % (min_value, max_value))
do_scaling = True
image.setProcessor(processor.convertToByte(do_scaling))
print("before gray morphology")
assert structuring_element_radius < 11, "the radius of the structuring element is too big (%d); using it with Fiji's 'Gray Morphology' tool would result in very long computations." % structuring_element_radius
imagej_run_image_command(image, "Gray Morphology", "radius=%d type=%s operator=%s" % (structuring_element_radius, structuring_element_shape, operator))
print("after gray morphology")
class WhiteEstimator(object):
def __init__(self, open_size, close_size, average_size):
self.open_size = open_size
self.close_size = close_size
self.average_size = average_size
def _remove_particles(self, white_estimate):
perform_gray_morphology(white_estimate, operator='open', structuring_element_shape='square', structuring_element_radius=(self.open_size + 1) / 2)
perform_gray_morphology(white_estimate, operator='close', structuring_element_shape='square', structuring_element_radius=(self.close_size + 1) / 2)
def estimate_white(self, sequences, channel_ids, dark=None):
"""Estimation of the white fluorescence image shape of synchrotron light from experimental images of Telemos microscope.
:param list(Sequence) sequences: the sequences to consider
:param list(str) channel_ids: the channels to consider, eg ['DM300_327-353_fluo']
:param WhiteEstimatorSettings white_estimator_settings:
:param ImagePlus or None dark:
:rtype ImagePlus:
Code adapted from matlab telemosToolbx's estimatedwhiteFluoImageTelemos function
% this function is specific of Telemos images (DISCO line SOLEIL)
% acquired using micromanager software linked to imageJ
%% input
% nothing : interactive function
%% output
% nothing : the final reference fluorescence image is saved on disk
%% principe
% NB: MICROMANAGER save images in a structured file folder architecture :
%
% root folder : name given by the user
%
% subfolders roi(n)_tile1 : n folders for each selected roi
% or pos(n) : n folders for each selected position
%
% display_and_comments.txt : file describing the channels acquired
%
% in each subfolder roi(n)_tile1 or Pos(n) :
% images files with name img_00000000(n)_DM300_327-353_00(p).tif
% n = number of time
% p = z focal plane
% DM300_327-353 = name of the channel
% metadata.txt : files describing all metadata associated with the
% acquisition : x, y, z position, camera settings .... etc.
%
% (Nb: depending on the date of acquisition, an extension fluo is found or
% not in the name of fluorescence image. This extension is given in
% contrast to the visible image.)
%
% IMAGES : fluorescence images are considered
%
%
% IMPORTANT:
% it is expected that in the folders at least several images cover the whole field of view
% illuminated by the synchrtron light
% IF IT IS NOT THE CASE, IT WON'T WORK
%
% BASIC: the estimated white image is the MAX of the selected images
% when interactive mode is selected, only first time and first z are
% proposed
% for automatic computing, all images are taken into account
%
% WHAT IS DONE:
% 0 - the user chosse the channels that are considered in the estimation of the
% white fluorescence image
%
% 1 - spurious pixels (three lines and columns around the image) are not taken
% in filtering and replaced by line and column number 4 and end-3
%
% 2 - Filtering
% the estimated dark image is filtered :
% opening to remove white regions brighter than the low frequency signal that should correspond to synchrotron light shape (should be small)
% closing to remove black regions that remains (should be small to avoid synchrotron light shape deformation)
% average filtering
% the image computed should be used to
% - to find the best white z plane from the reference white stack
% acquired for ex. using the so called "Matthieu lame"%
% - correct images for intensities
%% use
% estimatedwhiteFluoImageTelemos
%% Comments
% adapted from proposals 20161050 and 20171187
%% Author
% MF Devaux
% INRA BIA
% PVPP
%% date
% 5 octobre 2017:
% 23 mars 2018
% 3 septembre 2018 : comments and general case
% 16 avril 2019: name of function and default filtering values
% 29 avril 2019 : new comments and spurious pixels
% 27 mai 2019 : offset dark
% 4 juin 2019 : replace exist by isfolder or isfile
% 14 juin : close figure at the end
"""
images_file_path = []
for sequence in sequences:
for channel_id in channel_ids:
channel_index = sequence.get_channel_index(channel_id)
for frame_index in range(sequence.num_frames):
for z_index in range(sequence.num_slices):
images_file_path.append(sequence.get_image_file_path(channel_index, frame_index, z_index))
white_estimate = compute_max(images_file_path)
# modify spurious pixels on the side of the images
try:
replace_border(white_estimate, 3)
except NotImplementedError as error:
print('warning: replace_outer_frame is not implemented yet')
self._remove_particles(white_estimate)
return white_estimate
class InteractiveWhiteEstimator(WhiteEstimator):
def __init__(self, open_size, close_size, average_size):
WhiteEstimator.__init__(self, open_size, close_size, average_size)
def _remove_particles(self, white):
"""shows the image to a user so that he can visually check that the particles have been removed correctly in the given image
"""
raise NotImplementedError()
# part_rem_settings_are_good = False
# while part_rem_settings_are_good == False:
# # perform opening to remove white particles
# IJ.run( white_estimate, "Gray Morphology", "radius=1 type=square operator=open" )
# # run("Gray Morphology", "radius=1 type=square operator=open");
# # IJ.run( input_image_plus_copy, "Skeletonize (2D/3D)", "" )
# # perform
# if white_estimator_settings.particles_are_removed():
# part_rem_settings_are_good = true
# else
# white_estimator_settings.ask
def find_white_reference_image(white_estimate, white_z_stack):
"""Find the white reference Image among a z stack of reference image the most correlated to the white image estimated from a serie of acquisition.
:param Image white_estimate:
:param Sequence white_z_stack:
% this function is specific of Telemos images (DISCO line SOLEIL)
% acquired using micromanager software linked to imageJ
%% input
% nothing : interactive function
%% output
% zmax : focal plane
%% principe
% images were acquired for a given roi which position is found in
% metadata.txt file
% white and dark images were recorded for the full field of view of the DISCO
% TELEMOS camera
%
% correlation between a estimated white fluorescence image estalished from
% actual acquisitions of a given sample and all z plane acquired for the reference TELEMOS white
% image (Matthieu slide or any fluorescent homogeneous image) :
% The estimated white fluorescence image is generallly obtained bt using function whiteFluoImageTelemosestimation
% This is not compulsary as any homogenous sample image hat can roughly show the shape of illumination can be used to find
% the white reference image
%
%
% the z plane for which the maximum correlation is observed between estimated white and reference white images is retained.
% the white image is then offsetted (its corresponding Dark is subtracted) and copied in the subfolder <WhiteReference> of the sample
% rootfolder to show that it has been specifically selected for the considered experiment
%The matlab corrcoeff function is used
%
% correlation coefficients are saved in a file called
% 'corr.txt' in the subfolder 'WhiteReference'
%
%
% expected input folder hierarchy:
%
% >sampleFolder
% > PosFolders or RoiFolders
% > WhiteEstimate
% >darkFolder
% >darkFolder.smooth
% >whiteFolder
% > darkFolderforWhite
% > darkFolderforWhite.smooth
% >whiteFolder.smooth
%
%
% expected output folder hierarchy:
%
% >sampleFolder
% > PosFolders or RoiFolders
% > WhiteEstimate
% > WhiteReference
% > white after offset
% >darkFolder
% >darkFolder.smooth
% >whiteFolder
% > darkFolderforWhite
% > darkFolderforWhite.smooth
% >whiteFolder.smooth
%% use
% [zmax]=findWhiteReferenceImageTelemos
%% Comments
% written for proposal 20161050
% adapted for proposal 20170043
%% Author
% MF Devaux
% INRA BIA
% PVPP
%% date
% 5 octobre 2017:
% 15 decembre 2017 : adapted to take into account the roi known from
% metadata
% 27 fevrier 2018 : comments details
% 4 septembre 2018: comments and check and naming of folders
% 12 mars 2019 : save white reference with the same size as white estimate
% 16 avril 2019 : include diagonals to check the relevance of white
% reference
% 20 mai 2019 : track folder
% 27 mai 2019 : offset dark
% 4 juin 2019 : replace exist by isfolder or isfile
"""
raise NotImplementedError()
def test_preprocessing():
"""Test preprocessing."""
catalog = ImageCatalog('/Users/graffy/ownCloud/ipr/lipase/raw-images')
sequence = catalog.sequences['res_soleil2018/GGH/GGH_2018_cin2_phiG_I_327_vis_-40_1/Pos2']
white_estimator = WhiteEstimator(open_size=75, close_size=75, average_size=75)
white_estimate = white_estimator.estimate_white([sequence], ['DM300_327-353_fluo'])
find_white_reference_image(white_estimate, sequence.get_white())
if __name__ == '__main__':
test_preprocessing()