def configureTest():
"""
These tests have a lot of setup. This function takes care of this.
"""
mparams = params.ParametersMultiplane()
# sCMOS calibration files.
gain = numpy.ones(im_size)
offset = numpy.zeros(im_size)
variance = numpy.ones(im_size)
rqe = numpy.ones(im_size)
cal_file = storm_analysis.getPathOutputTest("c1_cal.npy")
numpy.save(cal_file, [offset, variance, gain, rqe, 2])
mparams.changeAttr("channel0_cal", cal_file)
cal_file = storm_analysis.getPathOutputTest("c2_cal.npy")
numpy.save(cal_file, [offset, variance, gain, rqe, 2])
mparams.changeAttr("channel1_cal", cal_file)
mparams.changeAttr("channel0_ext", "_c1.tif")
mparams.changeAttr("channel1_ext", "_c2.tif")
mparams.changeAttr("channel0_offset", 0)
mparams.changeAttr("channel1_offset", 0)
return mparams
def analyze(base_name, mlist_name, olist_name, settings_name):
parameters = params.ParametersMultiplane().initFromFile(settings_name)
finder = PFPeakFinder(parameters)
fitter = findPeaksStd.MPPeakFitter(parameters)
finder_fitter = PFPeakFinderFitter(parameters, finder, fitter)
reader = PFMovieReader(base_name=base_name,
bin_name=olist_name,
parameters=parameters)
stdAnalysis.standardAnalysis(finder_fitter, reader, mlist_name, parameters)
def analyze(base_name, mlist_name, settings_name):
parameters = params.ParametersMultiplane().initFromFile(settings_name)
finder = findPeaksStd.MPPeakFinder(parameters)
fitter = findPeaksStd.MPPeakFitter(parameters)
finder_fitter = findPeaksStd.MPFinderFitter(parameters, finder, fitter)
reader = findPeaksStd.MPMovieReader(base_name = base_name,
parameters = parameters)
data_writer = findPeaksStd.MPDataWriter(data_file = mlist_name,
parameters = parameters)
stdAnalysis.standardAnalysis(finder_fitter,
reader,
data_writer,
parameters)
def splitPeaks(mlist_filename, params_filename):
parameters = params.ParametersMultiplane().initFromFile(params_filename)
# Load the plane to plane mapping data.
mappings = {}
if parameters.hasAttr("mapping"):
if os.path.exists(parameters.getAttr("mapping")):
with open(parameters.getAttr("mapping"), 'rb') as fp:
mappings = pickle.load(fp)
else:
print("No mapping file parameter, nothing to do")
# Load frame offset information.
frame_offsets = list(
map(parameters.getAttr, mpUtilC.getOffsetAttrs(parameters)))
print(frame_offsets)
# Load the molecule list.
ch0_data = readinsight3.loadI3File(mlist_filename)
# Map to other channels.
basename = mlist_filename[:-4]
channel = 1
m_key = "0_1_"
while (m_key + "x") in mappings:
chn_data = ch0_data.copy()
# Map x.
tx = mappings[m_key + "x"]
chn_data['x'] = tx[0] + ch0_data['x'] * tx[1] + ch0_data['y'] * tx[2]
# Map y.
ty = mappings[m_key + "y"]
chn_data['y'] = ty[0] + ch0_data['x'] * ty[1] + ch0_data['y'] * ty[2]
# Map frame.
chn_data[
'fr'] = ch0_data['fr'] - frame_offsets[0] + frame_offsets[channel]
with writeinsight3.I3Writer(basename + "_ch" + str(channel) +
".bin") as i3w:
i3w.addMolecules(chn_data)
channel += 1
m_key = "0_" + str(channel) + "_"
def analyze(base_name, mlist_name, settings_name):
parameters = params.ParametersMultiplane().initFromFile(settings_name)
# Set to only analyze 1 frame, 1 iteration of peak finding.
parameters.setAttr("max_frame", "int", 1)
parameters.setAttr("iterations", "int", 1)
# Analyze one frame.
finder = PFPeakFinder(parameters)
fitter = PFPeakFitter(parameters)
finder_fitter = findPeaksStd.MPFinderFitter(parameters, finder, fitter)
reader = findPeaksStd.MPMovieReader(base_name=base_name,
parameters=parameters)
stdAnalysis.standardAnalysis(finder_fitter, reader, mlist_name, parameters)
# Evaluate found vs fit parameters.
hi = utilC.getHeightIndex()
xi = utilC.getXCenterIndex()
yi = utilC.getYCenterIndex()
# First identify peak pairs.
fi_x = fitted_peaks[:, xi]
fi_y = fitted_peaks[:, yi]
fo_x = found_peaks[:, xi]
fo_y = found_peaks[:, yi]
dd = utilC.peakToPeakDist(fo_x, fo_y, fi_x, fi_y)
di = utilC.peakToPeakIndex(fo_x, fo_y, fi_x, fi_y)
print(numpy.mean(dd), fi_x.size, fo_x.size)
fo_h = []
fi_h = []
for i in range(dd.size):
if (dd[i] < 2.0):
fo_h.append(found_peaks[i, hi])
fi_h.append(fitted_peaks[di[i], hi])
fi_h = numpy.array(fi_h)
fo_h = numpy.array(fo_h)
print(numpy.mean(fi_h), fi_h.size)
print(numpy.mean(fo_h), fo_h.size)
print(numpy.mean(fi_h) / numpy.mean(fo_h))
def test_offset_2():
"""
Movie reading test with non-zero offset.
"""
# Create XML file and sCMOS calibration files.
mparams = configureTest()
xml_file = storm_analysis.getPathOutputTest("mp_analysis.xml")
mparams.changeAttr("channel1_offset", 1)
mparams.toXMLFile(xml_file)
# Create movies.
mv_file = storm_analysis.getPathOutputTest("mp_c1.tif")
with tifffile.TiffWriter(mv_file) as tf:
for i in range(10):
im = (i + 1) * numpy.ones(im_size, dtype=numpy.int16)
tf.save(im.astype(numpy.int16))
mv_file = storm_analysis.getPathOutputTest("mp_c2.tif")
with tifffile.TiffWriter(mv_file) as tf:
for i in range(10):
im = (i + 1) * numpy.ones(im_size, dtype=numpy.int16)
tf.save(im.astype(numpy.int16))
# Create and test MPMovieReader
mpmr = analysis_io.MPMovieReader(
base_name=os.path.join(storm_analysis.getPathOutputTest(), "mp"),
parameters=params.ParametersMultiplane().initFromFile(xml_file))
mpmr.setup(0)
[mx, my, ml] = mpmr.getFilmSize()
assert (mx == im_size[1])
assert (my == im_size[0])
assert (ml == 9)
# Check that all of the frames are the same.
for i in range(ml):
frames = mpmr.getFrames(i)
assert (numpy.allclose(frames[0], frames[1] - 1))
parser.add_argument(
'--photons',
dest='photons',
type=int,
required=True,
help="An estimate of the average number of photons in the localization."
)
parser.add_argument('--xml',
dest='xml',
type=str,
required=True,
help="The name of the settings xml file.")
args = parser.parse_args()
parameters = params.ParametersMultiplane().initFromFile(args.xml)
spline_file_names = []
for spline_attr in mpUtilC.getSplineAttrs(parameters):
spline_file_names.append(parameters.getAttr(spline_attr))
#
# FIXME: Variances or weights need to be adjusted in x, y based on
# the mapping. This won't make much difference for symmetric
# PSFs but it is currently incorrect for asymmetric PSFs.
#
variances = planeVariances(args.background, args.photons,
parameters.getAttr("pixel_size"),
spline_file_names)
weights = {
"bg": planeWeights(variances[0]),
def findOffsets(base_name,
params_file,
background_scale=4.0,
foreground_scale=1.0):
"""
The 'main' function of this module.
base_name - The basename for the group of movies.
params_file - An analysis XML file containing the details for this experiment.
background_scale - Features in the background change on this scale (in pixels)
or more slowly.
foreground_scale - Features that change on this scale are likely foreground.
Notes:
1. This only checks a limited range of offsets between the two channels.
2. This assumes that the movies are longer than just a few frames.
"""
n_tests = 10
search_range = 5
# Load parameters.
parameters = params.ParametersMultiplane().initFromFile(params_file)
# Load the movies from each camera.
n_channels = 0
movies = []
for ext in mpUtil.getExtAttrs(parameters):
movie_name = base_name + parameters.getAttr(ext)
movies.append(datareader.inferReader(movie_name))
n_channels += 1
print("Found", n_channels, "movies.")
# Load sCMOS calibration data.
offsets = []
gains = []
for calib_name in mpUtil.getCalibrationAttrs(parameters):
[offset, variance, gain,
rqe] = analysisIO.loadCMOSCalibration(parameters.getAttr(calib_name))
offsets.append(offset)
gains.append(1.0 / gain)
assert (len(offsets) == n_channels)
# Load the plane to plane mapping data & create affine transform objects.
mappings = {}
with open(parameters.getAttr("mapping"), 'rb') as fp:
mappings = pickle.load(fp)
atrans = []
for i in range(n_channels - 1):
xt = mappings["0_" + str(i + 1) + "_x"]
yt = mappings["0_" + str(i + 1) + "_y"]
atrans.append(affineTransformC.AffineTransform(xt=xt, yt=yt))
# Create background and foreground variance filters.
#
# FIXME: Is this right for movies that are not square?
#
[y_size, x_size] = movies[0].filmSize()[:2]
psf = dg.drawGaussiansXY((x_size, y_size),
numpy.array([0.5 * x_size]),
numpy.array([0.5 * y_size]),
sigma=background_scale)
psf = psf / numpy.sum(psf)
bg_filter = matchedFilterC.MatchedFilter(psf)
psf = dg.drawGaussiansXY((x_size, y_size),
numpy.array([0.5 * x_size]),
numpy.array([0.5 * y_size]),
sigma=foreground_scale)
psf = psf / numpy.sum(psf)
fg_filter = matchedFilterC.MatchedFilter(psf)
var_filter = matchedFilterC.MatchedFilter(psf * psf)
# Check background estimation.
if False:
frame = loadImage(movies[0], 0, offsets[0], gain[0])
frame_bg = estimateBackground(frame, bg_filter, fg_filter, var_filter)
with tifffile.TiffWriter("bg_estimate.tif") as tif:
tif.save(frame.astype(numpy.float32))
tif.save(frame_bg.astype(numpy.float32))
tif.save((frame - frame_bg).astype(numpy.float32))
votes = numpy.zeros((n_channels - 1, 2 * search_range + 1))
for i in range(n_tests):
print("Test", i)
# Load reference frame.
ref_frame = loadImage(movies[0], search_range + i, offsets[0], gain[0])
ref_frame_bg = estimateBackground(ref_frame, bg_filter, fg_filter,
var_filter)
ref_frame -= ref_frame_bg
# Load test frames and measure correlation.
for j in range(n_channels - 1):
best_corr = 0.0
best_offset = 0
for k in range(-search_range, search_range + 1):
test_frame = loadImage(movies[j + 1],
search_range + i + k,
offsets[j + 1],
gain[j + 1],
transform=atrans[j])
test_frame_bg = estimateBackground(test_frame, bg_filter,
fg_filter, var_filter)
test_frame -= test_frame_bg
test_frame_corr = numpy.sum(
ref_frame * test_frame) / numpy.sum(test_frame)
if (test_frame_corr > best_corr):
best_corr = test_frame_corr
best_offset = k + search_range
votes[j, best_offset] += 1
# Print results.
print("Offset votes:")
print(votes)
frame_offsets = [0]
frame_offsets += list(numpy.argmax(votes, axis=1) - search_range)
print("Best offsets:")
for i in range(n_channels):
print(str(i) + ": " + str(frame_offsets[i]))
# Create stacks with optimal offsets.
print("Saving image stacks.")
for i in range(n_channels):
with tifffile.TiffWriter("find_offsets_ch" + str(i) + ".tif") as tif:
for j in range(5):
if (i == 0):
frame = loadImage(movies[i],
search_range + frame_offsets[i] + j,
offsets[i], gain[i])
else:
frame = loadImage(movies[i],
search_range + frame_offsets[i] + j,
offsets[i],
gain[i],
transform=atrans[i - 1])
frame_bg = estimateBackground(frame, bg_filter, fg_filter,
var_filter)
frame -= frame_bg
tif.save(frame.astype(numpy.float32))
