在不同的照明模式下StayGold的光稳定性
Masahiko Hirano1,2, Yasuo Yonemaru3,4, Satoshi Shimozono1
1Laboratory for Cell Function Dynamics, RIKEN Center for Brain Science, 2-1 Hirosawa, Wako-City, Saitama, 351-0198, Japan.
Scientific reports
|March 6, 2024
概括
与其他光蛋白相比,StayGold在活细胞成像中提供了卓越的光稳定性. 它的性能在激光扫描共聚焦显微镜中因照明模式而异.
科学领域:
- 细胞和分子成像技术
- 生物光子学 生物光子学
- 显微镜技术 显微镜技术
背景情况:
- 光蛋白 (FPs) 是活细胞成像中的关键工具.
- 光稳定性是许多FP的关键限制,特别是在强光照明下.
- 现有的绿色发射FP如EGFP,mClover3和mNeonGreen具有不同的光稳定性.
研究的目的:
- 为了评估一种新鲜明亮的光蛋白的光稳定性,StayGold.
- 在不同的显微镜照明模式中,将StayGold的光稳定性与已知的绿色FP (EGFP,mClover3,mNeonGreen) 进行比较.
- 研究激光扫描共聚焦显微镜 (LSCM) 参数对FP光稳定性的影响.
主要方法:
- 在HeLa细胞中表达StayGold,EGFP,mClover3和mNeonGreen作为基因2B的融合.
- 在广场,结构化照明和各种单光束和多光束激光扫描共聚焦显微镜 (LSCM) 照明模式下对光稳定性的评估.
- 系统检查单束LSCM扫描模式及其对FP光漂白的影响.
主要成果:
- 在广场和结构化照明显微镜下,StayGold在一个数量级上表现出比EGFP,mClover3和mNeonGreen更大的光稳定性.
- 与多束LSCM相比,StayGold的光稳定性在单束LSCM中显著降低.
- 发现FP光稳定性,包括StayGold的光稳定性,对化学固定方法敏感.
结论:
- StayGold代表了光蛋白在活细胞成像应用中的光稳定性的重大进步.
- 优化LSCM扫描模式对于最大限度地提高像StayGold这样的光稳定FP的实用性至关重要.
- 需要进一步的研究才能充分了解StayGold的光漂白机制和固定灵敏度.
更多相关视频
相关概念视频
01:26Photochemical Electrocyclic Reactions: Stereochemistry
1.4K
The absorption of UV–visible light by conjugated systems causes the promotion of an electron from the ground state to the excited state. Consequently, photochemical electrocyclic reactions proceed via the excited-state HOMO rather than the ground-state HOMO. Since the ground- and excited-state HOMOs have different symmetries, the stereochemical outcome of electrocyclic reactions depends on the mode of activation; i.e., thermal or photochemical.
Selection Rules: Photochemical Activation
Selection Rules: Photochemical Activation
1.4K
01:23Photoluminescence: Fluorescence and Phosphorescence
5.6K
Photoluminescence is a process where a molecule absorbs light energy and re-emits it in the form of light. This phenomenon occurs when a substance absorbs photons, promoting its electrons to higher energy level excited states, followed by a relaxation process in which the electrons return to their original ground state energy levels and emit light. Photoluminescence is widely observed in various materials, including semiconductors, and organic and inorganic compounds.
A pair of electrons in a...
A pair of electrons in a...
5.6K
01:26Variables Affecting Phosphorescence and Fluorescence
3.9K
Fluorescence and phosphorescence are essential phenomena in fields like analytical chemistry, biological imaging, and materials science, where they detect molecular properties and visualize cellular structures. Understanding the variables that influence these luminescent behaviors is crucial for maximizing accuracy and efficiency in their applications. These variables can broadly be grouped into chemical structure, solvent properties, and external conditions, each playing a distinct role in...
3.9K
01:14Photoluminescence: Applications
1.3K
Photoluminescence offers a wide range of applications due to its inherent sensitivity and selectivity. This technique allows for both direct and indirect analyses of the analyte. Direct quantitative analysis is possible when the analyte exhibits a favorable quantum yield for fluorescence or phosphorescence. However, an indirect analysis may be feasible if the analyte is not fluorescent or phosphorescent, or if the quantum yield is unfavorable. Indirect methods include reacting the analyte with...
1.3K