The video demonstrates a cryosectioning technique for obtaining discrete dorsal root ganglion (DRG) tissue sections. It details the process of attaching DRG sections to glass slides without overlap, ensuring optimal placement for subsequent immunohistochemistry studies. The effectiveness of the method is confirmed through confocal fluorescence microscopy imaging.

cryosectioning of the dorsal root ganglion (drg)

小鼠背根神经节冷冻切片

The dorsal root ganglion (DRG) contains the primary sensory neurons, the tissue macrophages, and the satellite cells that surround the primary sensory neurons1,2,3,4. It is a key anatomic structure in processing innocuous and noxious signals, and plays critical roles in pain, itch, and various peripheral nerve disorders5,6,7,8,9,10,11,12,13. Although several methods have been developed to dissect DRG tissue from the mouse spinal cord14,15,16, cryosectioning of the DRG tissue remains challenging as the DRG tissue is quite small, and cryostat sections of DRG samples tend to curve into rolls, making it difficult to properly transfer the cryostat sections onto glass slides. However, proper cryosectioning of the DRG tissue is crucial for immunohistochemistry studies and the structure of DRG sensory neurons17,18,19,20,21,22,23. Moreover, as single-cell RNA sequencing results have revealed the remarkable heterogeneity of DRG sensory neurons in both humans24 and mice25, proper cryosectioning of DRG tissue is critical for investigating the functional role of different DRG cells in various physiological and pathological conditions.
Although the tissue-clearing technique has been applied to investigate the 3D reconstruction of the DRG26 as an alternative technique of cryosectioning the DRG, the tissue-clearing technique is time- and labor-consuming. In comparison, cryosectioning of the DRG is quick and relatively easy to perform, and thus it remains to be a key technique for immunohistochemistry and structure studies of the DRG and other regions of the central nervous system. However, obtaining high-quality, intact, and flat cryostat sections onto glass slides remains to be a challenge in neuroscience research because of the tiny sample size of tissues, like the DRG and certain brain regions, and there is no article describing the optimal protocol at this point for cryosectioning small-sized tissue samples, such as mouse DRGs.
This protocol provides an easy, step-by-step technique for cryostat sectioning of the mouse DRG to reliably obtain as many high-quality DRG sections on the slides for subsequent DRG studies. While specifically designed for cryosectioning DRG samples, this technique can potentially be used for cryosectioning various other tissues with a small sample size.

作者聚焦：小鼠背根神经节冷冻切片程序  

小鼠背根神经节冷冻切片的程序

Watch this Scientific Journal Video about Cryosectioning of the Dorsal Root Ganglion DRG at JoVE.com

Cryosectioning of the Dorsal Root Ganglion (DRG)  

背根神经节冷冻切片 （DRG）

使用小油漆刷在20摄氏度下预冷，将最佳切割温度DRG部分保持在底部，开始冷冻切片。使用室温镊子的末端轻轻触摸该部分的底部，使其粘在平台表面上。接下来，慢慢地将带电的载玻片放在该部分上。一旦该部分粘在滑轨上，轻轻地将滑轨向后拉。按照相同的过程将更多 DRG 部分放到幻灯片上，而不会与这些部分重叠。在组织不与其他 DRG 切片的 OCT 重叠的地方，可以看到载玻片上 DRG 切片的正确位置。共聚焦荧光显微镜图像证明了使用该协议获得的DRG切片的质量。通过这种技术，可以使用不同的抗体进行DRG的免疫组织化学研究。

cryosectioning-of-the-dorsal-root-ganglion-drg

Research

JoVE Journal

Neuroscience

Cryosectioning of the Dorsal Root Ganglion (DRG)

544 次观看. 使用小油漆刷在20摄氏度下预冷，将最佳切割温度DRG部分保持在底部，开始冷冻切片。使用室温镊子的末端轻轻触摸该部分的底部，使其粘在平台表面上。接下来，慢慢地将带电的载玻片放在该部分上。一旦该部分粘在滑轨上，轻轻地将滑轨向后拉。按照相同的过程将更多 DRG 部分放到幻灯片上，而不会与这些部分重叠。在组织不与其他 DRG 切片的 OCT 重叠的地方，可以?...

视频: Cryosectioning of the Dorsal Root Ganglion (DRG)  

A Procedure for Mouse Dorsal Root Ganglion Cryosectioning

High-quality mouse dorsal root ganglion (DRG) cryostat sections are crucial for proper immunochemistry staining and RNAscope studies in the research of inflammatory and neuropathic pain, itch, as well as other peripheral neurological conditions. However, it remains a challenge to consistently obtain high-quality, intact, and flat cryostat sections onto glass slides because of the tiny sample size of the DRG tissue. So far, there is no article describing an optimal protocol for DRG cryosectioning. This protocol presents a step-by-step method to resolve the frequently encountered difficulties associated with DRG cryosectioning. The presented article explains how to remove the surrounding liquid from the DRG tissue samples, place the DRG sections on the slide facing the same orientation, and flatten the sections on the glass slide without curving up. Although this protocol has been developed for cryosectioning the DRG samples, it can be applied for the cryosectioning of many other tissues with a small sample size.

Presented here is the development for consistently acquiring high-quality dorsal root ganglion cryostat sections.

High-quality mouse dorsal root ganglion (DRG) cryostat sections are crucial for proper immunochemistry staining and RNAscope studies in the research of ...