Cell Biology

Many interesting cellular structures are smaller than the optical diffraction limit of ~200-300 nm. These include the substructures of most organelles and all macromolecular machines, channels, and receptors.

询问和回答有关这些结构和其他结构的分子组织，相互作用和化学计量的问题，需要在低于光的衍射极限的分辨率下对标记结构进行成像。

Conventional Microscopy Techniques in Cell Biology

Conventional light microscopy methods, like widefield and confocal microscopy, can image and identify specifically labeled cellular structures. However, they are limited in their ability to image below the optical diffraction limit. Alternatively, electron microscopy (EM) can achieve resolutions down to 0.1-0.2 nm laterally, but specific labeling or quantitative measurement of molecules within the cell is very challenging.

细胞生物学的超分辨率荧光显微镜

超分辨率显微镜支持高分辨率成像 - 横向下至20 nm，沿光轴50 nm - 专门标记的细胞结构。因此，它弥合了常规光与电子显微镜（EM）技术之间的差距。

_{*右：用Alexa 647对TOM20染色的线粒体，用SMLM（DSTORM）成像。}

The high resolution (20 nm laterally) paired with the ability to image specifically labeled structures makes SMLM a powerhouse solution in cell biology research. Discoveries made with SMLM include the visualization of local movements of chromatin domains in HeLa cells, the eightfold symmetry within the nuclear pore complex of Xenopus oocytes and the radial ninefold symmetry of the centriolar protein CEP164, and the organization of connected tubular structures within the endoplasmic reticulum[1, 2].

适合SMLM研究的纳米级细胞结构的例子

超分辨率显微镜支持高分辨率成像 - 横向下至20 nm，沿光轴50 nm - 专门标记的细胞结构。因此，它弥合了常规光与电子显微镜（EM）技术之间的差距。

The resolution needed to visualize structures of interest must be taken into consideration when deciding if SMLM is the right solution. Some examples of the sizes of diffraction-limited structures suitable for imaging with SMLM are provided. Examples* include (but are not limited to):

• 核孔复合物的扩散通道（长度为40-50nm）
• 线粒体中的核苷（直径约100 nm）
• 微管（厚度为24 nm）
• Ribosome (~20-30 nm in diameter)
• Bacterial heat shock protein DegP (20 nm in diameter)
• 叶绿体中的圆柱形小管（直径为107±26 nm）
• 小鼠细胞中的内吞坑（直径86±2.4nm）
• 大鼠肝线粒体（直径30-40 nm）中Cristae的管状片段
• Plasma membrane caveolae (50-100 nm in diameter)
• 细菌鞭毛钩(~ 55纳米长度)

_{*纳米级子结构及其尺寸的示例来自“有用的生物学数据库”。关联：https://bionumbers.hms.harvard.edu/search.aspx}

在超分辨率显微镜方法中，单分子定位显微镜（SMLM）可以说是最适合细胞生物学研究的。

具体来说，SMLM is the best method for studying specific nanoscale structures within cells when the research question requires:

• 可视化分子相距至20 nm；
• 标记多个特定分子结构；或者
• 定量数据收集在此规模上。
  

SMLM已经被用来回答以前未开发的细胞生物学问题。SMLM是在3D和高分辨率和量化数据中成像多个特定结构的理想选择。请参阅下面的示例数据和描述，以了解有关SMLM可以唯一实现的研究应用程序的更多信息：

固定样品成像

使用原始/seconday抗体标记的Alexa 647标记的固定微管的VXL -DSTROM图像中获得的微管。

活细胞成像

Acquired on the VXL - Imaging of live BSC1 cells labeled with Alexa 647 transferrin.

Bruker的Vutara VXL提供了一流的易用性和成像深度。支持细胞生物学研究人员需求的关键特征包括：

• 自动化工作流：SRX软件使得可以轻松获取必要的数据集，并且工作流程使用户可以专注于生物学问题，而不是修补复杂的显微镜设置。
• Superior 3D Imaging and Sample Flexibility:专有双平面检测提供了三维信息，并在组织切片中进行成像。
• 无限的多路复用：The integrated Microfluidics Unit allows sequential labeling of an unlimited number of fluorescent probes.
• Expert Applications Support:Vutara VXL users can receive personalized guidance for sample prep optimization specific to their research objectives.