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高折射率成像缓冲器用于双色 3D SMLM 厚样品成像

Lulu Zhou¹, Wei Shi¹, Shuang Fu¹

¹Department of Biomedical Engineering, Southern University of Science and Technology, Shenzhen 518055, China.

Analytical chemistry

|September 19, 2024

概括

研究人员通过开发折射率 (RI) 匹配的缓冲器,通过单分子局部化显微镜 (SMLM) 改进了深层组织成像. 这一突破使得在更厚的生物样本中能够在没有专门设备的情况下实现高分辨率的SMLM.

科学领域:

生物物理学的生物物理.
显微镜技术的使用方法
光学成像技术的使用.

背景情况:

单分子定位显微镜 (SMLM) 面临深层组织成像方面的局限性,原因是深度依赖的异常.
样品和客观浸泡介质之间的折射率 (RI) 不匹配导致了这些偏差,阻碍了更深处的高分辨率成像.

研究的目的:

为了扩大SMLM的成像深度.
通过将其 RI 与目标沉浸介质相匹配来优化成像缓冲器.
评估RI匹配的缓冲对染料闪和超分辨率图像质量的影响.

主要方法:

对五种不同的RI匹配的SMLM成像缓冲器进行系统评估.
专注于红色吸收染料的闪行为.
在各种深度重建超分辨率图像质量的评估.
使用了基于尾酒的清晰无障碍脑成像缓冲器,与油RI相匹配.

主要成果:

确定了一种RI匹配的缓冲器,能够清除组织样本和匹配RI的油.
在微米到几十微米的深度展示了高质量的双色3D SMLM成像.
在培养细胞和切割组织样本中实现了更好的成像.

结论:

与RI匹配的成像缓冲器显著提高了SMLM透深度.
这种方法为高分辨率深层组织成像提供了一种实用且易于使用的方法.
不需要专门的光学设备或专业知识,使先进的成像更容易获得.

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