研究目的
To develop a chameleon sensor for live cell imaging of mitochondrial DNA (mtDNA) that can selectively bind to mtDNA and provide dual color imaging to study mtDNA aggregation and mitophagy in cells stressed with anticancer doxorubicin.
研究成果
The study successfully developed a chameleon sensor, SG-RB, for live cell imaging of mtDNA, demonstrating its utility in studying mtDNA aggregation and mitophagy. The sensor's ability to provide dual color imaging based on the cellular location of mtDNA offers a novel approach to investigate mtDNA dynamics in pathological conditions.
研究不足
The study primarily focuses on HeLa cells and the effects of doxorubicin, which may not represent all cell types or stressors. The sensor's performance in other cell types or under different conditions was not extensively explored.
1：Experimental Design and Method Selection:
The study involved the synthesis of SG-RB, a chameleon sensor combining a cationic rhodamine B entity for mitochondria targeting and a SYBR Green-I entity for DNA sensing. The fluorescence response of SG-RB to plasmid DNA was tested in vitro.
2：Sample Selection and Data Sources:
HeLa cells were used to test the sensor's ability to image mtDNA and mitophagy.
3：List of Experimental Equipment and Materials:
Confocal fluorescence microscopy was performed on Zeiss LSM
4：Experimental Procedures and Operational Workflow:
7 Cells were cultured with SG-RB and other dyes, then visualized by confocal microscopy. The effects of doxorubicin on mtDNA were also examined.
5：Data Analysis Methods:
Fluorescence emission was analyzed using SpectraMax M5, and images were processed using Zen 2.3.0.
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Zeiss LSM 780
LSM 780
Zeiss
Confocal fluorescence microscopic imaging
ZEISS LSM 990 Spectral Multiplex
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SpectraMax M5
M5
SpectraMax
Fluorescence spectra analysis
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SYBR Green-I
Bioluminor
Fluorogenic DNA sensing
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MitoTracker Deep Red
Thermo Fisher
Mitochondria staining
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LysoTracker Blue
Thermo Fisher
Lysosomes staining
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Hoechst
Thermo Fisher
Nucleus staining
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