研究目的
Developing a near-infrared two-photon fluorescent probe for the detection of CE2 with high selectivity and sensitivity to evaluate its real-time activity in living cells, cancer tissues, and colon carcinoma mice model.
研究成果
The NIR-emissive two-photon fluorescent probe DCM-CES2 successfully detected endogenous CE2 activity in living cells and in vivo, offering potential advantages for studying the biological function of CE2 in complex systems.
研究不足
The probe's application may be limited by its specificity to CE2 and the need for further validation in more complex biological systems.
1：Experimental Design and Method Selection:
The probe DCM-CES2 was designed based on the attachment of an enzyme-active moiety L-leucine to an excellent NIR TP chromophore dicyanomethylene-4H-pyran (DCM).
2：Sample Selection and Data Sources:
Human hepatocellular liver carcinoma cell line (HepG2) and human colorectal cancer cell line HT-29 were used.
3：List of Experimental Equipment and Materials:
UV-Vis absorption spectrum, fluorescence spectrum, TP microscope, IVIS Lumina Kinetic Series III imaging system.
4：Experimental Procedures and Operational Workflow:
The probe was incubated with CE2 in PBS buffer pH = 7.4, 10 mM at 37 °C. Fluorescence imaging was performed at different depths in mice colon cancer slices.
5：4, 10 mM at 37 °C. Fluorescence imaging was performed at different depths in mice colon cancer slices.
Data Analysis Methods:
5. Data Analysis Methods: The fluorescence intensity was measured and analyzed to determine the activity of CE2.
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