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在生物组织中使用MINFLUX光纳米镜
Thea Moosmayer1,2, Kamila A Kiszka1, Volker Westphal1
1Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Göttingen 37077, Germany.
概括
MINFLUX纳米镜能够在完整的大脑组织中实现纳米级蛋白质成像,克服了分散的挑战. 这种超分辨率技术精确地定位了组织深处的分子,使得详细的突触结构分析成为可能.
科学领域:
- 神经科学是一个神经科学.
- 生物物理学的生物物理.
- 光学显微镜的使用方法
背景情况:
- 在完整的组织中实现纳米级蛋白质分布成像对于理解生理上下文中的细胞功能至关重要.
- 超分辨率光显微镜在分散和吸收生物组织方面面临挑战,限制了分子定位精度.
- 假设MINFLUX纳米镜,利用一个共聚焦针孔,通过提高信号与背景比率来克服这些限制.
研究的目的:
- 评估MINFLUX纳米镜的性能,用于在小鼠大脑组织段中成像蛋白质分布.
- 评估单个光体在具有挑战性的组织环境和深度内的定位精度.
- 调查双色MINFLUX在同时可视化不同突触蛋白点方面的潜力.
主要方法:
- MINFLUX (最小发射检测) 纳米镜应用于固定小鼠大脑组织部分.
- 用特定的光探针标记不同的突触蛋白点.
- 进行两色成像,以分析多种蛋白质和细胞结构的相对定位.
主要成果:
- 单个光体在5nm以下的精度下被定位,即使在组织内深度高达80μm.
- 通过MINFLUX成像,可视化后突触密度蛋白95 (PSD95) 与脊柱头形态相对.
- 该技术成功地解决了囊泡性谷氨酸转运体1 (VGlut1) 和α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid受体 (AMPARs) 在突触后的聚类.
- 在双色实验中,在10nm以下的3D光体定位精度.
结论:
- 在分散的生物组织中,MINFLUX纳米镜显示了纳米级蛋白质成像的高精度和深度透.
- 该研究验证了MINFLUX在单突触水平上对突触蛋白组织的详细,多色可视化.
- 这种技术为推进固定和活脑切片中突触结构和功能的研究提供了显著的潜力.
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