研究目的
To develop a liquid crystal (LC)-based aptasensor for the detection of interferon-γ (IFN-γ) in human blood and its application in the diagnosis of tuberculosis (TB).
研究成果
The LC-based aptasensor developed in this study is a simple, rapid, label-free, and easy-to-fabricate tool for the detection of IFN-γ and the diagnosis of latent TB. It offers advantages over conventional methods, including high sensitivity and the absence of intricate processes or instruments.
研究不足
The study was limited by the number of clinical blood samples from patients with latent TB used for testing. Additionally, the sensitivity of the sensor, while high, may require further optimization for clinical applications.
1：Experimental Design and Method Selection:
The study utilized a liquid crystal (LC)-based aptasensor system where the binding of IFN-γ to an aptamer immobilized on a surface disrupts the orientation of LCs, inducing a transition from a homeotropic orientation to a random one. This change was observed under a polarized light microscope.
2：Sample Selection and Data Sources:
Blood samples from patients with latent TB were used after pre-stimulation with TB antigen to induce IFN-γ release.
3：List of Experimental Equipment and Materials:
4-Cyano-4′-pentylbiphenyl (5CB), polished silicon wafer, microscope glass slides, dimethyloctadecyl[3-(trimethoxysilyl)propyl]ammonium chloride (DMOAP), (3-aminopropyl)triethoxysilane (APTES), glutaraldehyde (GA) solution, IFN-γ, human serum albumin (HSA), bovine serum albumin (BSA), ethanolamine, Tween? 20, and phosphate-buffered saline.
4：Experimental Procedures and Operational Workflow:
The aptamer was immobilized on a surface treated with APTES/DMOAP. The binding of IFN-γ to the aptamer was detected by monitoring the optical responses of LCs under a polarized light microscope.
5：Data Analysis Methods:
The optical LC images were analyzed to determine the orientational transition of LCs. Surface analysis was performed using atomic force microscopy (AFM) and ellipsometry.
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Atomic force microscopy
NanoScope IIIa
Veeco Metrology
Used for surface analysis to confirm the binding of IFN-γ to the aptamer-immobilized surface.
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4-Cyano-4′-pentylbiphenyl
C1555
Tokyo Chemical Industry
Used as the liquid crystal material in the aptasensor.
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Polished silicon wafer
Silicon Sense
Used for precise measurements with AFM and ellipsometry.
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Microscope glass slides
S-1215
Matunami
Used as the substrate for the aptasensor.
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Dimethyloctadecyl[3-(trimethoxysilyl)propyl]ammonium chloride
Sigma-Aldrich
Used for surface treatment to introduce LCs with a homeotropic orientation.
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(3-aminopropyl)triethoxysilane
Sigma-Aldrich
Used for surface treatment to immobilize the aptamer.
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Glutaraldehyde solution
Sigma-Aldrich
Used to functionalize surfaces with aldehyde groups for aptamer immobilization.
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Interferon-γ
Sigma-Aldrich
The target molecule for detection by the aptasensor.
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Human serum albumin
Sigma-Aldrich
Used in control experiments to test the specificity of the aptasensor.
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Bovine serum albumin
Sigma-Aldrich
Used in control experiments to test the specificity of the aptasensor.
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Ethanolamine
Sigma-Aldrich
Used for blocking non-specific binding of protein to the surface.
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Tween? 20
Sigma-Aldrich
Used in the preparation of solutions for the aptasensor.
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Phosphate-buffered saline
Sigma-Aldrich
Used as a buffer in the aptasensor experiments.
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Polarizing light microscope
ECLIPSE LV100POL
Nikon
Used to observe the optical responses of LCs in the aptasensor.
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Digital camera
DS-2Mv
Nikon
Used to capture images of the LC responses under the microscope.
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Ellipsometer
Elli-SE(UV)-FM8
Ellipso Technology
Used to measure the optical thickness of the organic surface layer at each step of the experiment.
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