研究目的
To develop a novel chemiluminescent probe for selective detection of cysteine (Cys) in living systems, distinguishing it from other biothiols like homocysteine and glutathione, and to apply it for in vivo imaging.
研究成果
The probe CL-Cys effectively detects cysteine with high sensitivity and selectivity, enabling chemiluminescent imaging in living mice. It provides a non-invasive method for studying cysteine's roles in physiological and pathological processes, with potential for further biological applications.
研究不足
The probe may have limitations in specificity under certain biological conditions, and the synthesis yield was low (22% for CL-Cys). In vivo applications might be affected by tissue penetration and background signals, though chemiluminescence minimizes these issues.
1：Experimental Design and Method Selection:
The probe CL-Cys was designed based on the Schaap’s adamantylidene-dioxetane scaffold, utilizing a Michael addition reaction with Cys followed by intramolecular cyclization to generate chemiluminescence. The synthesis involved multiple steps including protection, oxidation, and acylation reactions. Spectroscopic measurements were conducted to assess chemiluminescent properties.
2：Sample Selection and Data Sources:
Cysteine and other analytes (e.g., GSH, Hcy) were used in aqueous solutions. In vivo experiments were performed on SCID/BALB-C mice.
3：List of Experimental Equipment and Materials:
Equipment included Varian INOVA 300 spectrometer, Bruker DMX-600 spectrometer, LTQ mass spectrometry, Ultra-Weak Luminescence Analyzer (BPCL-GP-TGC), Molecular Devices SpectraMax M3, ProteinSimple FlourChem for imaging, and IVIS system. Chemicals included Tris-HCl buffer, DMSO, and various reagents for synthesis.
4：Experimental Procedures and Operational Workflow:
The probe was synthesized, characterized by NMR and MS. Chemiluminescent responses were measured in buffer solutions with varying Cys concentrations, pH levels, and selectivity tests. Cytotoxicity was assessed using CCK assay. In vivo imaging involved subcutaneous injections of probe and Cys mixtures in mice, with chemiluminescence captured using imaging systems.
5：Data Analysis Methods:
Chemiluminescent intensity was quantified, linear relationships and detection limits were calculated, and statistical analysis was performed for repeated experiments.
独家科研数据包，助您复现前沿成果，加速创新突破
获取完整内容-
Spectrometer
Bruker DMX-600
Bruker
Used for measuring 13C NMR spectra.
-
Ultra-Weak Luminescence Analyzer
BPCL-GP-TGC
Used for acquiring chemiluminescent spectra and kinetic curves.
-
Spectrometer
Varian INOVA 300
Varian
Used for measuring 1H NMR spectra.
-
Mass Spectrometry
LTQ
Used for recording electrospray ionization mass spectra (ESI-MS).
-
Microplate Reader
Molecular Devices SpectraMax M3
Molecular Devices
Used for recording chemiluminescent emission spectrum.
-
Imaging System
ProteinSimple FlourChem
ProteinSimple
Used for bulk chemiluminescent imaging.
-
IVIS System
Used for in vivo imaging of chemiluminescence in living mice.
-
ULTRAPURE WATER system
Used for purifying ultrapure water for experiments.
-
登录查看剩余6件设备及参数对照表
查看全部
