使用 Jython 脚本分析 FRAP 电影

import java.awt.Color as Color
from ij import WindowManager as WindowManager
from ij.plugin.frame import RoiManager as RoiManager
from ij.process import ImageStatistics as ImageStatistics
from ij.measure import Measurements as Measurements
from ij import IJ as IJ
from ij.measure import CurveFitter as CurveFitter
from ij.gui import Plot as Plot
from ij.gui import PlotWindow as PlotWindow
import math

# Get ROIs
roi_manager = RoiManager.getInstance()
roi_list    = roi_manager.getRoisAsArray()

# We assume first one is FRAP roi, the 2nd one is normalizing roi.
roi_FRAP    = roi_list[0];
roi_norm    = roi_list[1];

# Specify up to what frame to fit and plot.
n_slices = 30

# Get current image plus and image processor
current_imp  = WindowManager.getCurrentImage()
stack        = current_imp.getImageStack()
calibration  = current_imp.getCalibration()

#############################################

# Collect intensity values

# Create empty lists of number
If = []  # Frap values
In = []  # Norm values

# Loop over each slice of the stack
for i in range(0, n_slices):
 
    # Get the current slice 
    ip = stack.getProcessor(i+1)
 
    # Put the ROI on it
    ip.setRoi(roi_FRAP)
 
    # Make a measurement in it
    stats = ImageStatistics.getStatistics(ip, Measurements.MEAN, calibration);
    mean  = stats.mean
 
    # Store the measurement in the list
    If.append( mean  )

    # Do the same for non-FRAPed area
    ip.setRoi(roi_norm)
    stats = ImageStatistics.getStatistics(ip, Measurements.MEAN, calibration);
    mean = stats.mean
    In.append( mean  )
 
# Gather image parameters
frame_interval = calibration.frameInterval
time_units = calibration.getTimeUnit()
IJ.log('For image ' + current_imp.getTitle() )
IJ.log('Time interval is ' + str(frame_interval) + ' ' + time_units)
 
# Find minimal intensity value in FRAP and bleach frame
min_intensity = min( If )
bleach_frame = If.index( min_intensity )
IJ.log('FRAP frame is ' + str(bleach_frame+1) + ' at t = ' + str(bleach_frame * frame_interval) + ' ' + time_units )
 
# Compute mean pre-bleach intensity
mean_If = 0.0
mean_In = 0.0
for i in range(bleach_frame):         # will loop until the bleach time
    mean_If = mean_If + If[i]
    mean_In = mean_In + In[i]
mean_If = mean_If / bleach_frame
mean_In = mean_In / bleach_frame
 
# Calculate normalized curve
normalized_curve = []
for i in range(n_slices):
    normalized_curve.append( (If[i] - min_intensity) / (mean_If - min_intensity)   *   mean_In / In[i] )
    
x = [i * frame_interval for i in range( n_slices ) ] 
y = normalized_curve

xtofit = [ i * frame_interval for i in range( n_slices - bleach_frame ) ]
ytofit = normalized_curve[ bleach_frame : n_slices ]
 
# Fitter
fitter = CurveFitter(xtofit, ytofit)
fitter.doFit(CurveFitter.EXP_RECOVERY_NOOFFSET)
IJ.log("Fit FRAP curve by " + fitter.getFormula() )
param_values = fitter.getParams()
IJ.log( fitter.getResultString() )
 
# Overlay fit curve, with oversampling (for plot)
xfit = [ (t / 10.0  + bleach_frame) * frame_interval for t in range(10 * len(xtofit) ) ]
yfit = []
for xt in xfit:
    yfit.append( fitter.f( fitter.getParams(), xt - xfit[0]) )

 
plot = Plot("Normalized FRAP curve for " + current_imp.getTitle(), "Time ("+time_units+')', "NU", [], [])
plot.setLimits(0, max(x), 0, 1.2 );
plot.setLineWidth(2)


plot.setColor(Color.BLACK)
plot.addPoints(x, y, Plot.LINE)
plot.addPoints(x,y,PlotWindow.X);

 
plot.setColor(Color.RED)
plot.addPoints(xfit, yfit, Plot.LINE)

plot.setColor(Color.black);
plot_window =  plot.show()


# Output FRAP parameters
thalf = math.log(2) / param_values[1]
mobile_fraction = param_values[0]

str1 = ('Half-recovery time = %.2f ' + time_units) % thalf
IJ.log( str1 )
str2 = "Mobile fraction = %.1f %%" % (100 * mobile_fraction)
IJ.log( str2 )