This profile is often used
in a typical SPR experiment to indicate that the metal film (or
the solution above the metal film) matches the SPR resonance condition,
and that the apparatus is set up properly. However, the actual measurement
of molecular adsorption kinetics is only related to the shift of
the absorption (SPR) dip corresponding to the SPR angle change (Figure
2). One has to sacrifice the resolution of the measurement
by acquiring the whole intensity profile curves and comparing them
to find the shift of the dip (minimum).
Click
Here to View Animation Figure 2.
A light is beamed upon a metal film through a prism and the reflected
beam image shows a dark line due to SPR. The intensity profile of
the reflected beam exhibits a dip or minimal intensity at the resonance
angle. An SPR experiment measures the position shift of the dip
(the angle shift) upon molecular adsorption, and this shift represents
the adsorption kinetics when plotted as a function of time.
Biosensing Instrument Incorporated uses a different approach to
detect the SPR angle change by using a position-sensitive detector
to detect the SPR absorption dip shift. Since it only measures
the position shift of the dip, it offers a highly sensitive detection
scheme to measure extremely small angle changes of the SPR. As
long as the SPR angle resides inside the angle spread, the system
delivers exceptionally high angular resolution in its measurement.
Even though intensity profile is commonly used for demonstrating
the proper alignment and the setup of an SPR system, it only offers
limited status check of a metal film at a single position. For
a dual-channel detection scheme, a single intensity profile check
is not sufficient to ensure that the system is in proper SPR detection
condition. One has to ensure that all the illuminated areas of
the metal film are in good condition. BI-SPR instruments provides
the user a simple but sufficient solution by projecting the SPR
absorption dip onto a screen that can be visualized. By examining
the quality of the dark line (as shown in figure 3-5),
one could ensure that all conditions of the SPR are met and a
successful experiment can be conducted. |
As illustrated in Figure
3, a good metal film gives a well-defined, clean, and straight
dark line. This guarantees a good intensity profile at any point
along the dark line. Figure 4 and 5 demonstrated
that a partially contaminated metal film can still give a reasonable
intensity profile of SPR at a given point. However, it will not
lead to a reliable measurement, since the film quality across a
large area is bad.
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