Optical biosensor assay (OBA)

1991 ◽  
Vol 37 (9) ◽  
pp. 1502-1505 ◽  
Author(s):  
Y G Tsay ◽  
C I Lin ◽  
J Lee ◽  
E K Gustafson ◽  
R Appelqvist ◽  
...  

Abstract We describe a new biosensor immunoassay involving optical diffraction to detect clinically important analytes in human body fluids. A silicon wafer is used as a support for immobilization of antigen or antibody. The protein-coated surface is illuminated through a photo mask to create distinct periodic areas of active and inactive protein. When the surface is incubated with a positive sample, antigen-antibody binding occurs only on the active areas. Upon illumination with a light source such as a laser, the resulting biological diffraction grating diffracts the light. A negative sample does not result in diffraction because no antigen-antibody binding occurs to create the diffraction grating. The presence or absence of a diffraction signal differentiates between positive and negative samples, and the intensity of the signal provides a quantitative measure of the analyte concentration. The technique is demonstrated with a quantitative assay of choriogonadotropin in serum.

1983 ◽  
Vol 157 (6) ◽  
pp. 2121-2139 ◽  
Author(s):  
J Michl ◽  
M M Pieczonka ◽  
J C Unkeless ◽  
G I Bell ◽  
S C Silverstein

We describe a method for synchronously assembling antigen-antibody complexes underneath macrophages adherent to an antigen-coated surface. We have used this method to study the mechanism of Fc receptor (FcR) disappearance that occurs when resident and thioglycollate-elicited mouse macrophages are cultured on immune complex-coated surfaces. Erythrocytes opsonized with IgG (E(IgG) and a monoclonal antibody (2.4G2 IgG) directed against the trypsin-resistant FcR (FcRII) were used as indicators of the presence and distribution of FcRII molecules on the macrophage plasma membrane. Inhibitors of aerobic (NaCN) and anerobic (2-deoxyglucose, NaF) glycolysis and pinocytosis, of protein biosynthesis (cycloheximide), and of cytoskeletal function (cytochalasin B and D, colchicine, podophyllotoxin, taxol) did not reduce the rate or extent of FcRII modulation. Moreover, treatment of the macrophages with 0.1-0.5% formaldehyde did not reduce the extent of FcRII modulation as measured by the disappearance of E(IgG) binding sites. FcRII modulation was markedly slowed when the temperature was decreased to 2-4 degrees C. These results prove that FcRII modulation is governed by diffusion of the receptor in the plasma membrane. From the speed of FcRII disappearance from the macrophage's upper surface we calculate that the receptor has a diffusion coefficient at 37 degrees C of 2.5 X 10(-9) cm2/s. This finding indicates that FcRII, in its unligated form, is not linked to the macrophage's cytoskeleton, and that the receptor is capable of accommodating spatially to any distribution of ligands on a particle's surface.


2005 ◽  
Vol 295-296 ◽  
pp. 189-194 ◽  
Author(s):  
G.H. Wu ◽  
Z.J. Cai ◽  
Li Jiang Zeng

A two-color heterodyne interferometer based on the movement of the optical diffraction grating is proposed. The method allows us to measure the phase of synthetic wavelength f s directly and with high accuracy to extend the range of unambiguity for interferometric measurements by using two close wavelengths. Our experiment results show that the uncertainty in displacement measurement caused by the uncertainty in f s is 0.20 µm, smaller than the half of a single wavelength we used. The fringe order of a single wavelength can be determined without ambiguity. The uncertainty in displacement measurement can be improved further by using a single wavelength.


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