Aptamer LC (liquid crystal) biosensor for detecting pulmonary surfactant protein A as well as preparation and detection method of aptamer LC biosensor
A nucleic acid aptamer, lung surfactant technology, applied in the field of detection, to achieve the effect of rapid and efficient detection
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Embodiment 1
[0038] Determination of the optimal ratio of APTES / DMOAP on the bottom slide
[0039] In order to ensure the vertical uniform orientation of the liquid crystal, and to ensure the highest possible sensitivity of the sensor, the influence of the mixing ratio of APTES / DMOAP on the orientation of the liquid crystal was investigated successively. The test results are detailed in the appendix of the manual. Figure 4 . When the volume ratio of APTES / DMOAP is 5:1, the image appears uniform black (see the appendix Figure 4 a), when the volume ratio of APTES / DMOAP is increased to 10:1, the image still presents a uniform black color (see the appendix Figure 4 b), when the volume ratio of the two continues to increase to 15:1, the Schlelen texture begins to appear in the image (see the attached manual Figure 4 c), when the volume ratio is further increased to 20:1, the image appears obvious Schlelen texture (see the appendix Figure 4 d), in order to ensure that as many nucleic aci...
Embodiment 2
[0041] Optimal concentration of aptamer (Aptamer) on the surface of slides
[0042] The amino-modified nucleic acid aptamer was immobilized on the surface of an aldylated glass slide, and it was predicted that the concentration of Aptamer would affect the orientation of the liquid crystal molecules, (see the appendix for details). Figure 5 ) SP-A Aptamer liquid crystal optical signal map) Therefore, it is necessary to further determine the appropriate concentration of Aptamer immobilized on the glass surface. When the concentration of Aptamer is high (500 nmol·L -1 , 300 nmol L -1 , 200 nmol L -1 ), due to the excessive amount of Aptamer fixed on the surface of the glass slide, it has a great influence on the orientation of liquid crystal molecules, resulting in large colored and bright areas in the image (see the appendix Figure 5 a, b, c), in order to ensure that the subsequent detection is not affected, the concentration of Aptamer needs to be reduced. When the concent...
Embodiment 3
[0044] Investigation of sensor sensitivity
[0045] Different concentrations of pulmonary surfactant protein A (SP-A) solutions were dropped on the lower glass slide immobilized with nucleic acid aptamers, and after treatment, they were assembled face-to-face with the DMOAP-modified upper glass slide to form a liquid crystal cell. Observation, the observation results are detailed in the appendix of the instruction manual Image 6 . The higher the concentration of pulmonary surfactant protein A, the more it binds to Aptamer, so the greater the damage to the vertical orientation of liquid crystal molecules, the more obvious the colored bright areas and Schlelen texture in the image, when the pulmonary surfactant protein The concentration of A is 200nmol·L -1 When the color texture optical image is seen under the polarizing microscope (see the appendix Image 6 a); when the concentration of pulmonary surfactant protein A decreased to 100 nmol L -1 , 20 nmol L -1 and 10 nmol ...
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