In: Physics
Treat the two channel Young
The interferometric biosensor, which is a single-sensor,
real-time optical measurement
device targeted for label-free protein detection and analysis, has
been developed.
This is the compound result of various improvements to the optical
construction, such
as the introduction of the beam-forming optics and the adjustment
of the double-slit,
as well as its secondary use as a spatial filter at the input
coupling of the optical grating
for the successful reduction of scattering.
The optical biochips supplied by Unaxis optics have undergone
measurement and examination
in order to determine their operation characteristics. It is
concluded that the
addition of protective layers above the grating couplers is an
acceptable solution for
the prevention of the unwanted measurement of the silicon flow
cells’ refractive index.
Even with the protective layers, the optical coupling efficiency is
more than sufficient
for measurements. Furthermore, the scattering phenomena resulting
from foreign material
on the waveguide surface and their disruptive effects have been
acceptably
compensated by the aforementioned double-slit solution.
The software of the interferometric biosensor has been enhanced and
expanded into a
stable software-measurement system. There are, in fact, two
available LabVIEW-based
measurement programs, which each use different fundamental signal
analysis algorithms:
FFT-based and Fourier-based correlation. Each of these programs
contains a full
compliment of signal processing methods (i.e. window functions and
zero padding)
and noise averaging algorithms (i.e. block averaging and
moving-average filtering).
As for the signal analysis algorithms themselves, there is no
conclusive evidence to the
support the superiority of either algorithm. This is mainly based
on a lack of comparative
data and the existence of yet unexplained phenomena (e.g.
oscillations in the
phase trend and a large drift) for the Fourier-based correlation
algorithm.
Measurements and simulations have shown that the implementation of
specialized window
functions offers no justifiable improvement to the quality of the
final measurement
signal. Furthermore, windowing is a method for reducing the random
noise component
of the overall noise if and only if its implementation increases
the SNR enough to
compensate for its own contribution to the random noise component,
or uncertainty.
Zero padding as a linear interpolation method also offers no
justifiable improvement
to the quality of the final measurement signal, since the
interferogram period precision
without spectral linear interpolation is sufficient for the
analysis of the interferogram’s
phase. Thus, the use of other, possibly more sophisticated
interpolation methods is also
unjustified. Therefore, zero padding should remain a method for the
fulfillment of the
requirements, such that the signal analysis can be performed with
the more efficient
FFT algorithm.
The exact nature and components of the noise in the measurement
signal has not
been conclusively determined, but rather significantly narrowed
down to a few probabilities.
The nature of the noise has been shown to be statistical in nature,
without the
presence of periodicity and with the small presence of a random
noise component.
Theoretically, the major contributer to the noise and fluctuations
of the measurement
signal is the fluctuation in the external propagation path
difference. Calculations, simulations,
and observations support this, but the findings are not yet
conclusive.
With noise averaging it has been shown to be possible to reduce the
noise present in
the measurement signal, but averaging is not an elimination method.
Measurements
and simulations with block averaging suggest a noise minimum at a
CCD camera exposure
time of nearly 50 ms. This may perhaps be uncharacteristic of other
CCD camera
applications due to the interim FFT analysis needed to arrive at
the measurement
signal and due to the presence of operating system conflicts at
exposure times lower
than 40-60 ms. Based on this and the need for an acceptable
resolution of the measurement
signal, a system time constant has been recommended in the range
between
200 ms and 600 ms, assuming the use of block averaging factors
between 5
and 10. Similar measurements and simulations justify the additional
use of smoothing
algorithms based on a moving-average filter. Due to the
subjectivity of their usage a
conclusive smoothing factor has not been suggested, however a
factor range from
16-64 will result in noise reductions around a factor of 5 or
greater.
The improvement of the interferometric biosensor and its final
operation have been
tracked and recorded through the execution of numerous
glycerin-based tests. Several
remaining areas of improvement would be the signal drift and the
continual elimination
of noise sources and reduction of noise in the measurement signal.
Again, despite
exhaustive measurements and simulations, it has not yet been
possible to correlate the
drift to any single source or any combination of sources. The
current drift factor remains
near 5.0 ×10−6 ne f f /hour, which is larger than the interim goal
of 1.0 ×10−6 ne f f /hour.
The layering of the test sample on the surface of the waveguide has
proved to be a
promising hypothesis for the drift, but only based on observation.
Therefore, the cause
of the drift remains inconclusive.