def makeData(cal_file="calib.npy", dither=False, verbosity=1):
index = 1
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3: background.UniformBackground(
s, x, y, i3, photons=bg)
cam_f = lambda s, x, y, i3: camera.SCMOS(s, x, y, i3, cal_file)
pp_f = lambda s, x, y, i3: photophysics.AlwaysOn(s, x, y, i3, photons)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(s, x, y, i3, settings.
pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
dither=dither,
x_size=settings.x_size,
y_size=settings.y_size)
sim.simulate(wdir + "/test.tif",
"grid_list.hdf5",
settings.n_frames,
verbosity=verbosity)
index += 1
makeDataCommon.makePeakFile(settings)
def makeSampleData():
# Create sample bead data for fiducial tracking.
#
# Create randomly located localizations file.
#
print("Creating random localizations.")
sim_path = os.path.dirname(inspect.getfile(storm_analysis)) + "/simulator/"
subprocess.call([
"python", sim_path + "emitters_uniform_random.py", "--bin",
"random.hdf5", "--density",
str(density), "--margin",
str(margin), "--sx",
str(x_size), "--sy",
str(y_size)
])
# Create X/Y/Z drift file.
#
dx = numpy.random.normal(loc=x_loc, scale=x_scale, size=n_frames)
dy = numpy.random.normal(loc=y_loc, scale=y_scale, size=n_frames)
# Integrate dx, dy
for i in range(1, dx.size):
dx[i] = dx[i - 1] + dx[i]
dy[i] = dy[i - 1] + dy[i]
drift_data = numpy.zeros((dx.size, 3))
drift_data[:, 0] = dx
drift_data[:, 1] = dy
numpy.savetxt("drift.txt", drift_data)
# Create simulated data for fiducial tracking.
#
bg_f = lambda s, x, y, h5: background.UniformBackground(
s, x, y, h5, photons=10)
cam_f = lambda s, x, y, h5: camera.Ideal(s, x, y, h5, camera_offset)
drift_f = lambda s, x, y, h5: drift.DriftFromFile(s, x, y, h5, "drift.txt")
pp_f = lambda s, x, y, h5: photophysics.SimpleSTORM(
s, x, y, h5, 4000, on_time=10.0, off_time=1.0)
psf_f = lambda s, x, y, h5: psf.GaussianPSF(s, x, y, h5, pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
drift_factory=drift_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=x_size,
y_size=y_size)
sim.simulate("fiducials.tif", "random.hdf5", n_frames)
def test_simulate_1():
"""
No photo-physics, simple PSF, ideal camera.
"""
dax_name = storm_analysis.getPathOutputTest("test_sim1.dax")
bin_name = storm_analysis.getData("test/data/test_sim.hdf5")
sim = simulate.Simulate(background_factory = lambda settings, xs, ys, i3data : background.UniformBackground(settings, xs, ys, i3data),
camera_factory = lambda settings, xs, ys, i3data : camera.Ideal(settings, xs, ys, i3data, 100.0),
photophysics_factory = lambda settings, xs, ys, i3data : photophysics.AlwaysOn(settings, xs, ys, i3data, 1000.0),
psf_factory = lambda settings, xs, ys, i3data : psf.GaussianPSF(settings, xs, ys, i3data, 160.0),
x_size = 100, y_size = 75)
sim.simulate(dax_name, bin_name, 5)
def createMovie():
bg_f = lambda s, x, y, i3: background.UniformBackground(
s, x, y, i3, photons=bg)
cam_f = lambda s, x, y, i3: camera.Ideal(s, x, y, i3, camera_offset)
pp_f = lambda s, x, y, i3: photophysics.AlwaysOn(s, x, y, i3, signal)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(s, x, y, i3, pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=x_size,
y_size=y_size)
sim.simulate("test.tif", "sim_locs.hdf5", n_frames)
def createMovie(gain=1.0, offset=100.0):
bg_f = lambda s, x, y, h5: background.GaussianBackground(
s, x, y, h5, photons=bg)
cam_f = lambda s, x, y, h5: camera.Ideal(s, x, y, h5, offset, gain=gain)
pp_f = lambda s, x, y, h5: photophysics.SimpleSTORM(
s, x, y, h5, photons=signal)
psf_f = lambda s, x, y, h5: psf.GaussianPSF(s, x, y, h5, pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=x_size,
y_size=y_size)
sim.simulate("test.tif", "sim_locs.hdf5", n_frames)
def makeData(cal_file="calib.npy", dither=False):
index = 1
if True:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3: background.UniformBackground(
s, x, y, i3, photons=bg)
cam_f = lambda s, x, y, i3: camera.SCMOS(s, x, y, i3, cal_file)
pp_f = lambda s, x, y, i3: photophysics.AlwaysOn(
s, x, y, i3, photons)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(
s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
dither=dither,
x_size=settings.x_size,
y_size=settings.y_size)
sim.simulate(wdir + "/test.tif", "grid_list.hdf5",
settings.n_frames)
index += 1
# Create "peak_locations" file if needed.
#
if hasattr(settings, "peak_locations") and (settings.peak_locations
is not None):
with saH5Py.SAH5Py("test_01/test_ref.hdf5") as h5:
locs = h5.getLocalizationsInFrame(0)
if settings.peak_locations.endswith(".hdf5"):
saH5Py.saveLocalizations(settings.peak_locations, locs)
else:
numpy.savetxt(
settings.peak_locations,
numpy.transpose(
numpy.vstack((locs['x'], locs['y'], locs['height'],
locs['background']))))
def makeSampleData():
# Create sample bead data for fiducial tracking.
#
# Create randomly located localizations file.
#
print("Creating random localizations.")
emittersUniformRandom.emittersUniformRandom("random.hdf5", density, margin,
x_size, y_size, 0.0)
# Create X/Y/Z drift file.
#
dx = numpy.random.normal(loc=x_loc, scale=x_scale, size=n_frames)
dy = numpy.random.normal(loc=y_loc, scale=y_scale, size=n_frames)
# Integrate dx, dy
for i in range(1, dx.size):
dx[i] = dx[i - 1] + dx[i]
dy[i] = dy[i - 1] + dy[i]
drift_data = numpy.zeros((dx.size, 3))
drift_data[:, 0] = dx
drift_data[:, 1] = dy
numpy.savetxt("drift.txt", drift_data)
# Create simulated data for fiducial tracking.
#
bg_f = lambda s, x, y, h5: background.UniformBackground(
s, x, y, h5, photons=10)
cam_f = lambda s, x, y, h5: camera.Ideal(s, x, y, h5, camera_offset)
drift_f = lambda s, x, y, h5: drift.DriftFromFile(s, x, y, h5, "drift.txt")
pp_f = lambda s, x, y, h5: photophysics.SimpleSTORM(
s, x, y, h5, 4000, on_time=10.0, off_time=1.0)
psf_f = lambda s, x, y, h5: psf.GaussianPSF(s, x, y, h5, pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
drift_factory=drift_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=x_size,
y_size=y_size)
sim.simulate("fiducials.tif", "random.hdf5", n_frames)
def createMovie(n_frames):
bg_f = lambda s, x, y, i3: background.UniformBackground(
s, x, y, i3, photons=bg)
cam_f = lambda s, x, y, i3: camera.Ideal(s, x, y, i3, camera_offset)
pp_f = lambda s, x, y, i3: photophysics.AlwaysOn(s, x, y, i3, signal)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(s, x, y, i3, pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=x_size,
y_size=y_size)
sim.simulate("test.tif", "sim_locs.hdf5", n_frames)
# Also create file to use for peak locations.
with saH5Py.SAH5Py("test_ref.hdf5") as h5:
locs = h5.getLocalizationsInFrame(0)
saH5Py.saveLocalizations("peak_locs.hdf5", locs)
def makeData():
index = 1
# Gaussian non-uniform background, STORM.
if True:
for elt in ["drift_xy.txt", "drift_xyz.txt"]:
bg = settings.background
photons = settings.photons
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3: background.GaussianBackground(
s, x, y, i3, photons=bg)
cam_f = lambda s, x, y, i3: camera.Ideal(s, x, y, i3, settings.
camera_offset)
drift_f = lambda s, x, y, i3: drift.DriftFromFile(s, x, y, i3, elt)
pp_f = lambda s, x, y, i3: photophysics.SimpleSTORM(
s, x, y, i3, photons)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(
s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
drift_factory=drift_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=settings.x_size,
y_size=settings.y_size)
sim.simulate(wdir + "/test.dax", "clusters_list.hdf5",
settings.n_frames)
#sim.simulate(wdir + "/test.dax", "lines_list.hdf5", settings.n_frames)
index += 1
help="The length of the movie in frames.")
parser.add_argument('--photons',
dest='photons',
type=float,
required=True,
help="The integral of a single emitter in photons.")
args = parser.parse_args()
sim = Simulate(
lambda settings, xs, ys, h5data: background.UniformBackground(
settings, xs, ys, h5data), lambda settings, xs, ys, h5data: camera.
Ideal(settings, xs, ys, h5data, 100.0),
lambda settings, xs, ys, h5data: photophysics.AlwaysOn(
settings, xs, ys, h5data, args.photons),
lambda settings, xs, ys, h5data: psf.GaussianPSF(
settings, xs, ys, h5data, 160.0))
sim.simulate(args.dax_file, args.hdf5, args.frames)
#
# The MIT License
#
# Copyright (c) 2016 Zhuang Lab, Harvard University
#
# Permission is hereby granted, free of charge, to any person obtaining a copy
# of this software and associated documentation files (the "Software"), to deal
# in the Software without restriction, including without limitation the rights
# to use, copy, modify, merge, publish, distribute, sublicense, and/or sell
# copies of the Software, and to permit persons to whom the Software is
# furnished to do so, subject to the following conditions:
#
bg = settings.background
photons = settings.photons
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3: background.GaussianBackground(
s, x, y, i3, photons=bg)
cam_f = lambda s, x, y, i3: camera.Ideal(s, x, y, i3, settings.
camera_offset)
drift_f = lambda s, x, y, i3: drift.DriftFromFile(s, x, y, i3, elt)
pp_f = lambda s, x, y, i3: photophysics.SimpleSTORM(
s, x, y, i3, photons)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(s, x, y, i3, settings.
pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
drift_factory=drift_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=settings.x_size,
y_size=settings.y_size)
sim.simulate(wdir + "/test.dax", "clusters_list.hdf5",
settings.n_frames)
#sim.simulate(wdir + "/test.dax", "lines_list.hdf5", settings.n_frames)
index += 1
def configure():
# Get relevant paths.
mm_path = os.path.dirname(inspect.getfile(storm_analysis)) + "/micrometry/"
mp_path = os.path.dirname(
inspect.getfile(storm_analysis)) + "/multi_plane/"
sp_path = os.path.dirname(inspect.getfile(storm_analysis)) + "/spliner/"
# Create analysis XML files.
#
print("Creating XML files.")
params = testingParametersSCMOS()
params.toXMLFile("scmos.xml")
params = testingParametersMC()
params.toXMLFile("multicolor.xml")
# Useful variables
aoi_size = int(settings.psf_size / 2) + 1
# Create sCMOS data and HDF5 files we'll need for the simulation.
#
if True:
# Create sCMOS camera calibration files.
#
numpy.save("calib.npy", [
numpy.zeros(
(settings.y_size, settings.x_size)) + settings.camera_offset,
numpy.ones(
(settings.y_size, settings.x_size)) * settings.camera_variance,
numpy.ones(
(settings.y_size, settings.x_size)) * settings.camera_gain, 1
])
# Create localization on a grid file.
#
print("Creating gridded localizations.")
sim_path = os.path.dirname(
inspect.getfile(storm_analysis)) + "/simulator/"
subprocess.call([
"python", sim_path + "emitters_on_grid.py", "--bin",
"grid_list.hdf5", "--nx",
str(settings.nx), "--ny",
str(settings.ny), "--spacing", "20", "--zrange",
str(settings.test_z_range), "--zoffset",
str(settings.test_z_offset)
])
# Create randomly located localizations file (for STORM movies).
#
print("Creating random localizations.")
subprocess.call([
"python", sim_path + "emitters_uniform_random.py", "--bin",
"random_storm.hdf5", "--density", "1.0", "--margin",
str(settings.margin), "--sx",
str(settings.x_size), "--sy",
str(settings.y_size), "--zrange",
str(settings.test_z_range)
])
# Create randomly located localizations file (for mapping measurement).
#
print("Creating random localizations.")
subprocess.call([
"python", sim_path + "emitters_uniform_random.py", "--bin",
"random_map.hdf5", "--density", "0.0003", "--margin",
str(settings.margin), "--sx",
str(settings.x_size), "--sy",
str(settings.y_size)
])
# Create sparser grid for PSF measurement.
#
print("Creating data for PSF measurement.")
sim_path = os.path.dirname(
inspect.getfile(storm_analysis)) + "/simulator/"
subprocess.call([
"python", sim_path + "emitters_on_grid.py", "--bin",
"psf_list.hdf5", "--nx", "6", "--ny", "3", "--spacing", "40"
])
## This part makes / tests measuring the mapping.
##
if True:
print("Measuring mapping.")
# Make localization files for simulations.
#
locs = saH5Py.loadLocalizations("random_map.hdf5")
locs["z"][:] = 1.0e-3 * settings.z_planes[0]
saH5Py.saveLocalizations("c1_random_map.hdf5", locs)
for i in range(1, 4):
locs["x"] += settings.dx
locs["y"] += settings.dy
locs["z"][:] = settings.z_planes[i]
saH5Py.saveLocalizations("c" + str(i + 1) + "_random_map.hdf5",
locs)
# Make localization files for simulations.
#
locs = saH5Py.loadLocalizations("random_map.hdf5")
locs["z"][:] = 1.0e-3 * settings.z_planes[0]
saH5Py.saveLocalizations("c1_random_map.hdf5", locs)
for i in range(1, 4):
locs["x"] += settings.dx
locs["y"] += settings.dy
locs["z"][:] = settings.z_planes[i]
saH5Py.saveLocalizations("c" + str(i + 1) + "_random_map.hdf5",
locs)
# Make simulated mapping data.
#
bg_f = lambda s, x, y, h5: background.UniformBackground(
s, x, y, h5, photons=10)
cam_f = lambda s, x, y, h5: camera.SCMOS(s, x, y, h5, "calib.npy")
pp_f = lambda s, x, y, h5: photophysics.AlwaysOn(s, x, y, h5, 20000.0)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(s, x, y, i3, settings.
pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=settings.x_size,
y_size=settings.y_size)
for i in range(4):
sim.simulate("c" + str(i + 1) + "_map.dax",
"c" + str(i + 1) + "_random_map.hdf5", 1)
# Analyze simulated mapping data
#
for i in range(4):
scmos.analyze("c" + str(i + 1) + "_map.dax",
"c" + str(i + 1) + "_map.hdf5", "scmos.xml")
# Measure mapping.
#
for i in range(3):
subprocess.call([
"python", mm_path + "micrometry.py", "--locs1", "c1_map.hdf5",
"--locs2", "c" + str(i + 2) + "_map.hdf5", "--results",
"c1_c" + str(i + 2) + "_map.map", "--no_plots"
])
# Merge mapping.
#
subprocess.call([
"python", mm_path + "merge_maps.py", "--results", "map.map",
"--maps", "c1_c2_map.map", "c1_c3_map.map", "c1_c4_map.map"
])
# Print mapping.
#
if True:
print("Mapping is:")
subprocess.call([
"python", mp_path + "print_mapping.py", "--mapping", "map.map"
])
print("")
# Check that mapping is close to what we expect (within 5%).
#
with open("map.map", 'rb') as fp:
mappings = pickle.load(fp)
for i in range(3):
if not numpy.allclose(mappings["0_" + str(i + 1) + "_x"],
numpy.array(
[settings.dx * (i + 1), 1.0, 0.0]),
rtol=0.05,
atol=0.05):
print("X mapping difference for channel", i + 1)
if not numpy.allclose(mappings["0_" + str(i + 1) + "_y"],
numpy.array(
[settings.dy * (i + 1), 0.0, 1.0]),
rtol=0.05,
atol=0.05):
print("Y mapping difference for channel", i + 1)
## This part measures / test the PSF measurement.
##
if True:
# Create drift file, this is used to displace the localizations in the
# PSF measurement movie.
#
dz = numpy.arange(-settings.psf_z_range, settings.psf_z_range + 0.05,
0.01)
drift_data = numpy.zeros((dz.size, 3))
drift_data[:, 2] = dz
numpy.savetxt("drift.txt", drift_data)
# Also create the z-offset file.
#
z_offset = numpy.ones((dz.size, 2))
z_offset[:, 1] = dz
numpy.savetxt("z_offset.txt", z_offset)
# Create simulated data for PSF measurements.
#
bg_f = lambda s, x, y, h5: background.UniformBackground(
s, x, y, h5, photons=10)
cam_f = lambda s, x, y, h5: camera.SCMOS(s, x, y, h5, "calib.npy")
drift_f = lambda s, x, y, h5: drift.DriftFromFile(
s, x, y, h5, "drift.txt")
pp_f = lambda s, x, y, h5: photophysics.AlwaysOn(s, x, y, h5, 20000.0)
psf_f = lambda s, x, y, h5: psf.PupilFunction(s, x, y, h5, settings.
pixel_size, [])
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
drift_factory=drift_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=settings.x_size,
y_size=settings.y_size)
if True:
for i in range(4):
sim.simulate("c" + str(i + 1) + "_zcal.dax",
"c" + str(i + 1) + "_random_map.hdf5", dz.size)
# Get localizations to use for PSF measurement.
#
subprocess.call([
"python", mp_path + "psf_localizations.py", "--bin",
"c1_map_ref.hdf5", "--map", "map.map", "--aoi_size",
str(aoi_size)
])
# Create PSF z stacks.
#
for i in range(4):
subprocess.call([
"python", mp_path + "psf_zstack.py", "--movie",
"c" + str(i + 1) + "_zcal.dax", "--bin",
"c1_map_ref_c" + str(i + 1) + "_psf.hdf5", "--zstack",
"c" + str(i + 1) + "_zstack", "--scmos_cal", "calib.npy",
"--aoi_size",
str(aoi_size)
])
# Measure PSF.
#
for i in range(4):
subprocess.call([
"python", mp_path + "measure_psf.py", "--zstack",
"c" + str(i + 1) + "_zstack.npy", "--zoffsets", "z_offset.txt",
"--psf_name", "c" + str(i + 1) + "_psf_normed.psf",
"--z_range",
str(settings.psf_z_range), "--normalize"
])
## This part creates the splines.
##
if True:
print("Measuring Splines.")
for i in range(4):
subprocess.call([
"python", sp_path + "psf_to_spline.py", "--psf",
"c" + str(i + 1) + "_psf_normed.psf", "--spline",
"c" + str(i + 1) + "_psf.spline", "--spline_size",
str(settings.psf_size)
])
## This part measures the Cramer-Rao weights.
##
if True:
print("Calculating weights.")
subprocess.call([
"python", mp_path + "plane_weighting.py", "--background",
str(settings.photons[0][0]), "--photons",
str(settings.photons[0][1]), "--output", "weights.npy", "--xml",
"multicolor.xml", "--no_plots"
])
def makeData():
index = 1
# sCMOS calibration file.
if settings.random_variance:
variance = numpy.random.exponential(scale=settings.camera_variance,
size=(settings.y_size,
settings.x_size))
offset = settings.camera_offset + variance
numpy.save("calib.npy", [
offset, variance,
numpy.ones(
(settings.y_size, settings.x_size)) * settings.camera_gain, 1
])
else:
numpy.save("calib.npy", [
numpy.zeros(
(settings.y_size, settings.x_size)) + settings.camera_offset,
numpy.ones(
(settings.y_size, settings.x_size)) * settings.camera_variance,
numpy.ones(
(settings.y_size, settings.x_size)) * settings.camera_gain, 1
])
# sCMOS camera movies.
#
# For these simulations we expect (approximately) these results:
#
# Analysis Summary:
# Total analysis time 10.64 seconds
# Recall 0.93726
# Noise 0.05972
# XY Error (nm):
# test_01 14.44 14.53
# test_02 8.26 8.24
#
# XY Width Error, Mean difference with truth, Standard deviation (pixels):
# test_01 0.029 0.116 0.029 0.116
# test_02 0.017 0.105 0.017 0.105
#
if True:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3: background.UniformBackground(
s, x, y, i3, photons=bg)
cam_f = lambda s, x, y, i3: camera.SCMOS(s, x, y, i3, "calib.npy")
pp_f = lambda s, x, y, i3: photophysics.AlwaysOn(
s, x, y, i3, photons)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(
s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=settings.x_size,
y_size=settings.y_size)
sim.simulate(wdir + "/test.tif", "grid_list.hdf5",
settings.n_frames)
index += 1
def makeDataSpliner():
index = 1
# Non-uniform background, STORM.
#
if False:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3: background.GaussianBackground(
s, x, y, i3, photons=bg)
cam_f = lambda s, x, y, i3: camera.Ideal(s, x, y, i3, settings.
camera_offset)
pp_f = lambda s, x, y, i3: photophysics.SimpleSTORM(
s, x, y, i3, photons)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(
s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=settings.x_size,
y_size=settings.y_size)
sim.simulate(wdir + "/test.dax", "random_list.hdf5",
settings.n_frames)
index += 1
# Ideal camera movies, PSF using the measured spline.
#
if False:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3: background.UniformBackground(
s, x, y, i3, photons=bg)
cam_f = lambda s, x, y, i3: camera.Ideal(s, x, y, i3, settings.
camera_offset)
pp_f = lambda s, x, y, i3: photophysics.AlwaysOn(
s, x, y, i3, photons)
psf_f = lambda s, x, y, i3: psf.Spline(s, x, y, i3, settings.
pixel_size, "psf.spline")
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=settings.x_size,
y_size=settings.y_size)
sim.simulate(wdir + "/test.dax", "grid_list.hdf5",
settings.n_frames)
index += 1
# Create "peak_locations" file if needed.
#
if hasattr(settings, "peak_locations") and (settings.peak_locations
is not None):
with saH5Py.SAH5Py("test_01/test_ref.hdf5") as h5:
locs = h5.getLocalizationsInFrame(0)
if settings.peak_locations.endswith(".hdf5"):
saH5Py.saveLocalizations(settings.peak_locations, locs)
else:
numpy.savetxt(
settings.peak_locations,
numpy.transpose(
numpy.vstack((locs['x'], locs['y'], locs['height'],
locs['background']))))
def configure(no_splines, cal_file = None):
# Create sCMOS calibration file if requested.
#
if cal_file is not None:
offset = numpy.zeros((settings.y_size, settings.x_size)) + settings.camera_offset
variance = numpy.ones((settings.y_size, settings.x_size))
gain = numpy.ones((settings.y_size, settings.x_size)) * settings.camera_gain
rqe = numpy.ones((settings.y_size, settings.x_size))
numpy.save(cal_file, [offset, variance, gain, rqe, 2])
# Create parameters file for analysis.
#
print("Creating XML file.")
params = testingParameters(cal_file = cal_file)
params.toXMLFile("spliner.xml")
# Create localization on a grid file.
#
print("Creating gridded localization.")
sim_path = os.path.dirname(inspect.getfile(storm_analysis)) + "/simulator/"
subprocess.call(["python", sim_path + "emitters_on_grid.py",
"--bin", "grid_list.hdf5",
"--nx", str(settings.nx),
"--ny", str(settings.ny),
"--spacing", "20"])
# Create randomly located localizations file.
#
print("Creating random localization.")
subprocess.call(["python", sim_path + "emitters_uniform_random.py",
"--bin", "random_list.hdf5",
"--density", "1.0",
"--margin", str(settings.margin),
"--sx", str(settings.x_size),
"--sy", str(settings.y_size)])
# Create sparser grid for PSF measurement.
#
print("Creating data for PSF measurement.")
sim_path = os.path.dirname(inspect.getfile(storm_analysis)) + "/simulator/"
subprocess.call(["python", sim_path + "emitters_on_grid.py",
"--bin", "sparse_list.hdf5",
"--nx", "6",
"--ny", "3",
"--spacing", "40"])
if no_splines:
return
# Create simulated data for PSF measurement.
#
bg_f = lambda s, x, y, i3 : background.UniformBackground(s, x, y, i3, photons = 10)
cam_f = lambda s, x, y, i3 : camera.Ideal(s, x, y, i3, 100.)
pp_f = lambda s, x, y, i3 : photophysics.AlwaysOn(s, x, y, i3, 20000.0)
psf_f = lambda s, x, y, i3 : psf.GaussianPSF(s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory = bg_f,
camera_factory = cam_f,
photophysics_factory = pp_f,
psf_factory = psf_f,
dither = True,
x_size = settings.x_size,
y_size = settings.y_size)
sim.simulate("spline_2d.tif", "sparse_list.hdf5", 5)
# Measure the PSF.
#
print("Measuring PSF.")
spliner_path = os.path.dirname(inspect.getfile(storm_analysis)) + "/spliner/"
subprocess.call(["python", spliner_path + "measure_psf.py",
"--movie", "spline_2d.tif",
"--bin", "spline_2d_ref.hdf5",
"--psf", "psf.psf",
"--want2d",
"--aoi_size", str(settings.spline_size+1)])
# Measure the Spline.
#
if True:
print("Measuring Spline.")
subprocess.call(["python", spliner_path + "psf_to_spline.py",
"--psf", "psf.psf",
"--spline", "psf.spline",
"--spline_size", str(settings.spline_size)])
def configure(no_splines, cal_file = None):
# Create sCMOS calibration file if requested.
#
if cal_file is not None:
offset = numpy.zeros((settings.y_size, settings.x_size)) + settings.camera_offset
variance = numpy.ones((settings.y_size, settings.x_size))
gain = numpy.ones((settings.y_size, settings.x_size)) * settings.camera_gain
rqe = numpy.ones((settings.y_size, settings.x_size))
numpy.save(cal_file, [offset, variance, gain, rqe, 2])
# Create parameters file for analysis.
#
print("Creating XML file.")
params = testingParameters(cal_file = cal_file)
params.toXMLFile("spliner.xml")
# Create localization on a grid file.
#
print("Creating gridded localization.")
emittersOnGrid.emittersOnGrid("grid_list.hdf5",
settings.nx,
settings.ny,
1.5,
20,
0.0,
0.0)
# Create randomly located localizations file.
#
print("Creating random localization.")
emittersUniformRandom.emittersUniformRandom("random_list.hdf5",
1.0,
settings.margin,
settings.x_size,
settings.y_size,
0.0)
# Create sparser grid for PSF measurement.
#
print("Creating data for PSF measurement.")
emittersOnGrid.emittersOnGrid("sparse_list.hdf5",
6,
3,
1.5,
40,
0.0,
0.0)
if no_splines:
return
# Create beads.txt file for spline measurement.
#
with saH5Py.SAH5Py("sparse_list.hdf5") as h5:
locs = h5.getLocalizations()
numpy.savetxt("beads.txt", numpy.transpose(numpy.vstack((locs['x'], locs['y']))))
# Create simulated data for PSF measurement.
#
bg_f = lambda s, x, y, i3 : background.UniformBackground(s, x, y, i3, photons = 10)
cam_f = lambda s, x, y, i3 : camera.Ideal(s, x, y, i3, 100.)
pp_f = lambda s, x, y, i3 : photophysics.AlwaysOn(s, x, y, i3, 20000.0)
psf_f = lambda s, x, y, i3 : psf.GaussianPSF(s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory = bg_f,
camera_factory = cam_f,
photophysics_factory = pp_f,
psf_factory = psf_f,
dither = True,
x_size = settings.x_size,
y_size = settings.y_size)
sim.simulate("spline_2d.tif", "sparse_list.hdf5", 5)
# Measure the PSF.
#
print("Measuring PSF.")
psf_name = "psf.psf"
measurePSF.measurePSF("spline_2d.tif",
"na",
"sparse_list.hdf5",
psf_name,
want2d = True,
aoi_size = int(settings.spline_size + 1),
pixel_size = settings.pixel_size * 1.0e-3)
# Measure the Spline.
#
if True:
print("Measuring Spline.")
psfToSpline.psfToSpline(psf_name, "psf.spline", settings.spline_size)
def makeData(dither = False):
index = 1
# Gaussian PSF, uniform background.
if True:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3 : background.UniformBackground(s, x, y, i3, photons = bg)
cam_f = lambda s, x, y, i3 : camera.Ideal(s, x, y, i3, settings.camera_offset)
pp_f = lambda s, x, y, i3 : photophysics.AlwaysOn(s, x, y, i3, photons)
psf_f = lambda s, x, y, i3 : psf.GaussianPSF(s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory = bg_f,
camera_factory = cam_f,
photophysics_factory = pp_f,
psf_factory = psf_f,
dither = dither,
x_size = settings.x_size,
y_size = settings.y_size)
sim.simulate(wdir + "/test.dax", "grid_list.hdf5", settings.n_frames)
index += 1
# Pupil Function PSF.
if False:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3 : background.UniformBackground(s, x, y, i3, photons = bg)
cam_f = lambda s, x, y, i3 : camera.Ideal(s, x, y, i3, settings.camera_offset)
pp_f = lambda s, x, y, i3 : photophysics.AlwaysOn(s, x, y, i3, photons)
psf_f = lambda s, x, y, i3 : psf.PupilFunction(s, x, y, i3, settings.pixel_size, [])
sim = simulate.Simulate(background_factory = bg_f,
camera_factory = cam_f,
photophysics_factory = pp_f,
psf_factory = psf_f,
x_size = settings.x_size,
y_size = settings.y_size)
sim.simulate(wdir + "/test.dax", "grid_list.hdf5", settings.n_frames)
index += 1
# Gaussian non-uniform background, always on.
if False:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3 : background.GaussianBackground(s, x, y, i3, photons = bg)
cam_f = lambda s, x, y, i3 : camera.Ideal(s, x, y, i3, settings.camera_offset)
pp_f = lambda s, x, y, i3 : photophysics.AlwaysOn(s, x, y, i3, photons)
psf_f = lambda s, x, y, i3 : psf.GaussianPSF(s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory = bg_f,
camera_factory = cam_f,
photophysics_factory = pp_f,
psf_factory = psf_f,
x_size = settings.x_size,
y_size = settings.y_size)
sim.simulate(wdir + "/test.dax", "grid_list.hdf5", settings.n_frames)
index += 1
# Uniform background, STORM.
if False:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3 : background.UniformBackground(s, x, y, i3, photons = bg)
cam_f = lambda s, x, y, i3 : camera.Ideal(s, x, y, i3, settings.camera_offset)
pp_f = lambda s, x, y, i3 : photophysics.SimpleSTORM(s, x, y, i3, photons)
psf_f = lambda s, x, y, i3 : psf.GaussianPSF(s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory = bg_f,
camera_factory = cam_f,
photophysics_factory = pp_f,
psf_factory = psf_f,
x_size = settings.x_size,
y_size = settings.y_size)
sim.simulate(wdir + "/test.dax", "random_list.hdf5", settings.n_frames)
index += 1
# Gaussian non-uniform background, STORM.
if False:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3 : background.GaussianBackground(s, x, y, i3, photons = bg)
cam_f = lambda s, x, y, i3 : camera.Ideal(s, x, y, i3, settings.camera_offset)
pp_f = lambda s, x, y, i3 : photophysics.SimpleSTORM(s, x, y, i3, photons)
psf_f = lambda s, x, y, i3 : psf.GaussianPSF(s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory = bg_f,
camera_factory = cam_f,
photophysics_factory = pp_f,
psf_factory = psf_f,
x_size = settings.x_size,
y_size = settings.y_size)
sim.simulate(wdir + "/test.dax", "random_list.hdf5", settings.n_frames)
index += 1
# Sloped non-uniform background, always on.
if False:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3 : background.SlopedBackground(s, x, y, i3, slope = 0.4, offset = 10)
cam_f = lambda s, x, y, i3 : camera.Ideal(s, x, y, i3, settings.camera_offset)
pp_f = lambda s, x, y, i3 : photophysics.AlwaysOn(s, x, y, i3, photons)
psf_f = lambda s, x, y, i3 : psf.GaussianPSF(s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory = bg_f,
camera_factory = cam_f,
photophysics_factory = pp_f,
psf_factory = psf_f,
dither = True,
x_size = settings.x_size,
y_size = settings.y_size)
sim.simulate(wdir + "/test.dax", "grid_list.hdf5", settings.n_frames)
index += 1
# Sine non-uniform background, always on.
if False:
for [bg, photons] in settings.photons:
wdir = "test_{0:02d}".format(index)
print(wdir)
if not os.path.exists(wdir):
os.makedirs(wdir)
bg_f = lambda s, x, y, i3 : background.SineBackground(s, x, y, i3, photons = bg, period = 45)
cam_f = lambda s, x, y, i3 : camera.Ideal(s, x, y, i3, settings.camera_offset)
pp_f = lambda s, x, y, i3 : photophysics.AlwaysOn(s, x, y, i3, photons)
psf_f = lambda s, x, y, i3 : psf.GaussianPSF(s, x, y, i3, settings.pixel_size)
sim = simulate.Simulate(background_factory = bg_f,
camera_factory = cam_f,
photophysics_factory = pp_f,
psf_factory = psf_f,
dither = True,
x_size = settings.x_size,
y_size = settings.y_size)
sim.simulate(wdir + "/test.dax", "grid_list.hdf5", settings.n_frames)
index += 1
# Create "peak_locations" file if needed.
#
if hasattr(settings, "peak_locations") and (settings.peak_locations is not None):
with saH5Py.SAH5Py("test_01/test_ref.hdf5") as h5:
locs = h5.getLocalizationsInFrame(0)
if settings.peak_locations.endswith(".hdf5"):
saH5Py.saveLocalizations(settings.peak_locations, locs)
else:
numpy.savetxt(settings.peak_locations,
numpy.transpose(numpy.vstack((locs['x'],
locs['y'],
locs['height'],
locs['background']))))
def configure():
# Create analysis XML files.
#
print("Creating XML files.")
params = testingParametersSCMOS()
params.toXMLFile("scmos.xml")
params = testingParametersMC()
params.toXMLFile("multicolor.xml")
# Useful variables
aoi_size = int(settings.psf_size / 2) + 1
# Create sCMOS data and HDF5 files we'll need for the simulation.
#
if True:
# Create sCMOS camera calibration files.
#
numpy.save("calib.npy", [
numpy.zeros(
(settings.y_size, settings.x_size)) + settings.camera_offset,
numpy.ones(
(settings.y_size, settings.x_size)) * settings.camera_variance,
numpy.ones(
(settings.y_size, settings.x_size)) * settings.camera_gain,
numpy.ones((settings.y_size, settings.x_size)), 2
])
# Create localization on a grid file.
#
print("Creating gridded localizations.")
emittersOnGrid.emittersOnGrid("grid_list.hdf5", settings.nx,
settings.ny, 1.5, 20,
settings.test_z_range,
settings.test_z_offset)
# Create randomly located localizations file (for STORM movies).
#
print("Creating random localizations.")
emittersUniformRandom.emittersUniformRandom("random_storm.hdf5", 1.0,
settings.margin,
settings.x_size,
settings.y_size,
settings.test_z_range)
# Create randomly located localizations file (for mapping measurement).
#
print("Creating random localizations.")
emittersUniformRandom.emittersUniformRandom("random_map.hdf5", 0.0003,
settings.margin,
settings.x_size,
settings.y_size,
settings.test_z_range)
# Create sparser grid for PSF measurement.
#
print("Creating data for PSF measurement.")
emittersOnGrid.emittersOnGrid("psf_list.hdf5", 6, 3, 1.5, 40, 0.0, 0.0)
## This part makes / tests measuring the mapping.
##
if True:
print("Measuring mapping.")
# Make localization files for simulations.
#
locs = saH5Py.loadLocalizations("random_map.hdf5")
locs["z"][:] = 1.0e-3 * settings.z_planes[0]
saH5Py.saveLocalizations("c1_random_map.hdf5", locs)
for i in range(1, 4):
locs["x"] += settings.dx
locs["y"] += settings.dy
locs["z"][:] = settings.z_planes[i]
saH5Py.saveLocalizations("c" + str(i + 1) + "_random_map.hdf5",
locs)
# Make localization files for simulations.
#
locs = saH5Py.loadLocalizations("random_map.hdf5")
locs["z"][:] = 1.0e-3 * settings.z_planes[0]
saH5Py.saveLocalizations("c1_random_map.hdf5", locs)
for i in range(1, 4):
locs["x"] += settings.dx
locs["y"] += settings.dy
locs["z"][:] = settings.z_planes[i]
saH5Py.saveLocalizations("c" + str(i + 1) + "_random_map.hdf5",
locs)
# Make simulated mapping data.
#
bg_f = lambda s, x, y, h5: background.UniformBackground(
s, x, y, h5, photons=10)
cam_f = lambda s, x, y, h5: camera.SCMOS(s, x, y, h5, "calib.npy")
pp_f = lambda s, x, y, h5: photophysics.AlwaysOn(s, x, y, h5, 20000.0)
psf_f = lambda s, x, y, i3: psf.GaussianPSF(s, x, y, i3, settings.
pixel_size)
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=settings.x_size,
y_size=settings.y_size)
for i in range(4):
sim.simulate("c" + str(i + 1) + "_map.dax",
"c" + str(i + 1) + "_random_map.hdf5", 1)
# Analyze simulated mapping data
#
for i in range(4):
h5_name = "c" + str(i + 1) + "_map.hdf5"
if os.path.exists(h5_name):
os.remove(h5_name)
scmos.analyze("c" + str(i + 1) + "_map.dax", h5_name, "scmos.xml")
# Measure mapping.
#
for i in range(3):
micrometry.runMicrometry("c1_map.hdf5",
"c" + str(i + 2) + "_map.hdf5",
"c1_c" + str(i + 2) + "_map.map",
min_size=5.0,
max_size=100.0,
max_neighbors=20,
tolerance=1.0e-2,
no_plots=True)
# Merge mapping and save results.
#
merged_map = mergeMaps.mergeMaps(
["c1_c2_map.map", "c1_c3_map.map", "c1_c4_map.map"])
with open("map.map", 'wb') as fp:
pickle.dump(merged_map, fp)
# Print mapping.
#
if True:
print("Mapping is:")
printMapping.printMapping("map.map")
print("")
# Check that mapping is close to what we expect (within 5%).
#
with open("map.map", 'rb') as fp:
mappings = pickle.load(fp)
for i in range(3):
if not numpy.allclose(mappings["0_" + str(i + 1) + "_x"],
numpy.array(
[settings.dx * (i + 1), 1.0, 0.0]),
rtol=0.05,
atol=0.05):
print("X mapping difference for channel", i + 1)
if not numpy.allclose(mappings["0_" + str(i + 1) + "_y"],
numpy.array(
[settings.dy * (i + 1), 0.0, 1.0]),
rtol=0.05,
atol=0.05):
print("Y mapping difference for channel", i + 1)
## This part measures / test the PSF measurement.
##
if True:
# Create drift file, this is used to displace the localizations in the
# PSF measurement movie.
#
dz = numpy.arange(-settings.psf_z_range, settings.psf_z_range + 0.05,
0.01)
drift_data = numpy.zeros((dz.size, 3))
drift_data[:, 2] = dz
numpy.savetxt("drift.txt", drift_data)
# Also create the z-offset file.
#
z_offset = numpy.ones((dz.size, 2))
z_offset[:, 1] = dz
numpy.savetxt("z_offset.txt", z_offset)
# Create simulated data for PSF measurements.
#
bg_f = lambda s, x, y, h5: background.UniformBackground(
s, x, y, h5, photons=10)
cam_f = lambda s, x, y, h5: camera.SCMOS(s, x, y, h5, "calib.npy")
drift_f = lambda s, x, y, h5: drift.DriftFromFile(
s, x, y, h5, "drift.txt")
pp_f = lambda s, x, y, h5: photophysics.AlwaysOn(s, x, y, h5, 20000.0)
psf_f = lambda s, x, y, h5: psf.PupilFunction(s, x, y, h5, settings.
pixel_size, [])
sim = simulate.Simulate(background_factory=bg_f,
camera_factory=cam_f,
drift_factory=drift_f,
photophysics_factory=pp_f,
psf_factory=psf_f,
x_size=settings.x_size,
y_size=settings.y_size)
if True:
for i in range(4):
sim.simulate("c" + str(i + 1) + "_zcal.dax",
"c" + str(i + 1) + "_random_map.hdf5", dz.size)
# Get localizations to use for PSF measurement.
#
psfLocalizations.psfLocalizations("c1_map_ref.hdf5",
"map.map",
aoi_size=aoi_size)
# Create PSF z stacks.
#
for i in range(4):
psfZStack.psfZStack("c" + str(i + 1) + "_zcal.dax",
"c1_map_ref_c" + str(i + 1) + "_psf.hdf5",
"c" + str(i + 1) + "_zstack",
aoi_size=aoi_size)
# Measure PSF.
#
for i in range(4):
mpMeasurePSF.measurePSF("c" + str(i + 1) + "_zstack.npy",
"z_offset.txt",
"c" + str(i + 1) + "_psf_normed.psf",
z_range=settings.psf_z_range,
normalize=True)
## This part creates the splines.
##
if True:
print("Measuring Splines.")
for i in range(4):
psfToSpline.psfToSpline("c" + str(i + 1) + "_psf_normed.psf",
"c" + str(i + 1) + "_psf.spline",
int(settings.psf_size / 2))
## This part measures the Cramer-Rao weights.
##
if True:
print("Calculating weights.")
planeWeighting.runPlaneWeighting("multicolor.xml",
"weights.npy",
[settings.photons[0][0]],
settings.photons[0][1],
no_plots=True)
