研究目的
To visualize telomerase expression level in tumor cells by developing a plasmonic colorimetric sensor for highly sensitive detection of telomerase activity.
研究成果
The developed plasmonic colorimetric sensor offers a new opportunity for visualization and quantitative detection of telomerase activity with superior sensitivity and holds great potential to boost the visual detection of telomerase.
研究不足
The study focuses on the visual detection of telomerase activity in tumor cells, and the sensitivity is limited to 1 HeLa cell for LSPR peak measurement and a visual detection limit of 20 HeLa cells.
1:Experimental Design and Method Selection:
The study integrates an excellent etching substrate Au nanobipyramids (Au NBPs) with a liposome-based signal amplification strategy.
2:Sample Selection and Data Sources:
Telomerase extracted from a certain number of cells was serially diluted in 1×CHAPS lysis buffer.
3:List of Experimental Equipment and Materials:
Includes Au NBPs, HRP-encapsulated liposomes (HRP-Ls), magnetic beads, and H2O2-TMB substrate solution.
4:Experimental Procedures and Operational Workflow:
Telomerase activity was converted into the amount of the attached HRP-Ls on the surfaces of magnetic beads, followed by the liberation of HRP and initiation of the oxidation reaction between H2O2 and TMB to form TMB2+. The morphological evolution of Au NBPs was then observed.
5:2+. The morphological evolution of Au NBPs was then observed. Data Analysis Methods:
5. Data Analysis Methods: UV-vis absorption spectra of Au NBPs aqueous dispersion were measured under room temperature using a microplate spectrophotometer.
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Transmission electron microscopy
FEI Tecnai G2 F20
FEI
Used to observe the size and morphology of the Au NBPs.
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Dynamic light scattering system
Malvern Nano ZS90
Malvern
Used to measure the average size of the liposomes.
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Au nanobipyramids
Serving as the plasmonic signal readout element for the detection of telomerase activity.
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Horseradish peroxidase-encapsulated liposomes
Used for signal amplification in the detection of telomerase activity.
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Magnetic beads
Used to specifically harbor HRP-encapsulated liposomes under the action of telomerase.
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H2O2-3,3′,5,5′-tetramethylbenzidine sulfate
Used as a substrate for the oxidation reaction catalyzed by HRP.
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Microplate spectrophotometer
Synergy H1
BioTek
Used to measure UV-vis absorption spectra of Au NBPs aqueous dispersion.
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