Logic of this FWHM script? - matlab

Question

Could someone explain the logic of this program.
I dont understand why the y=y/max(y)
and,
interp = (0.5-y(i-1)) / (y(i)-y(i-1));
tlead = x(i-1) + interp*(x(i)-x(i-1));
The script:
function width = fwhm(x,y)
y = y / max(y);
N = length(y);
MicroscopeMag=10;
PixelWidth=7.8; % Pixel Pitch is 7.8 Microns.
%------- find index of center (max or min) of pulse---------------%
[~,centerindex] = max(y);% 479 S10 find center peak and coordinate
%------- find index of center (max or min) of pulse-----------------%
i = 2;
while sign(y(i)-0.5) == sign(y(i-1)-0.5) %trying to see the curve raise
i = i+1; %474 S10
end %first crossing is between v(i-1) & v(i)
interp = (0.5-y(i-1)) / (y(i)-y(i-1));
tlead = x(i-1) + interp*(x(i)-x(i-1));
i=centerindex+1; %471
%------- start search for next crossing at center--------------------%
while ((sign(y(i)-0.5) == sign(y(i-1)-0.5)) && (i <= N-1))
i = i+1;
end
if i ~= N
interp = (0.5-y(i-1)) / (y(i)-y(i-1));
ttrail = x(i-1) + interp*(x(i)-x(i-1));
%width = ttrail - tlead; % FWHM
width=((ttrail - tlead)/MicroscopeMag)*PixelWidth;
% Lateral Magnification x Pixel pitch of 7.8 microns.
end
Thanks.

Answer 1

The two segments of code you specifically mention are both housekeeping: it's more about the compsci of it than the optics.
So the first line
y = y/max(y);
is normalising it to 1, i.e. dividing the whole series through by the maximum value. This is a fairly common practice and it's sensible to do it here, it saves the programmer from having to divide through by it later.
The next part,
interp = (0.5-y(i-1)) / (y(i)-y(i-1));
tlead = x(i-1) + interp*(x(i)-x(i-1));
and the corresponding block later on for ttrail, are about trying to interpolate the exact point(s) where the signal's value would be 0.5. Earlier it identifies the centre of the peak and the last index position before half-maximum, so now we have a range containing the leading edge of the signal.
The 'half-maximum' criterion requires us to find the point where that leading edge's value is 0.5 (we normalised to 1, so the half-maximum is by definition 0.5). The data probably won't have a sample at exactly that value - it'll go [... 0.4856 0.5024 ...] or something similar.
So these two lines are an attempt to determine in fractions of an index exactly where the line would cross the 0.5 value. It does this by simple linear interpolation:
y(i)-y(i-1)
gives us the delta_y between the two values either side, and
0.5-y(i-1)
gives us the shortfall. By taking the ratio we can linearly interpolate how far between the two index positions we should go to hit exactly 0.5.
The next line then works out the corresponding delta_x, which gives you the actual distance in terms of the timebase.
It does the same thing for the trailing edge, then uses these two interpolated values to give you a more precise value for the full-width.
To visualise this I would put a breakpoint at the i = 2 line and step through it, noting or plotting the values of y(i) as you go. stem is helpful for visualising discrete data, especially when you're working between index positions.

Answer 2

The program computes the resolution of a microscope using the Full Width at Half Maximum (FWHM) of the Point Spread Function (PSF) characterizing the microscope with a given objective/optics/etc.
The PSF normally looks like a gaussian:
and the FWHM tells you how good is your microscope system to discern small objects (i.e. the resolution). Let's say you are looking at 2 point objects, then the resolution (indirectly FWHM) is the minimum size those objects need to be if you are indeed to tell that there are 2 objects close to one another instead of one big object.
Now for the above function, it looks like it first compute the maximum of the PSF and then progressively goes down along the curve until it approximately reaches the half maximum. Then it's possible to compute the FWHM from the distribution of the PSF.
Hope that makes things a bit clearer!