We present two innovations in histological technique for rodent spinal cord: gelatin embedding and LED photobleaching. Gelatin embedding uses liquid gelatin solution to permeate delicate biological structures then solidify to provide mechanical support throughout dissection, vibratome sectioning, and staining. LED photobleaching uses high-intensity visible light during blocking and primary incubations to reduce autofluorescence in tissue sections before fluorescent secondaries are added. We found gelatin embedding improved mechanical stability without interfering with immunohistochemical staining. Gelatin embedding also preserved some spinal roots and provided an opportunity for dye-less and cut-less tracking of left/right orientation during free-floating staining, which is valuable for tissue samples that have no spare areas that can be marked. LED photobleaching greatly reduced autofluorescence and added essentially no extra time or labor to the process. Descriptions of the techniques and characterization data are provided.

我们介绍了啮齿动物脊髓组织学技术的两项创新：明胶包埋和 LED 光漂白。明胶包埋使用液体明胶溶液渗透精细的生物结构，然后固化以在整个解剖、振动切片和染色过程中提供机械支撑。LED 光漂白在阻断和初级孵育期间使用高强度可见光，以在添加荧光二级之前减少组织切片中的自发荧光。我们发现明胶包埋提高了机械稳定性而不干扰免疫组织化学染色。明胶嵌入还保留了一些脊髓根，并提供了在自由浮动染色期间对左/右方向进行无染料和无切割跟踪的机会，这对于没有可标记的备用区域的组织样本很有价值。LED 光漂白大大减少了自发荧光，并且基本上不会在该过程中增加额外的时间或劳动力。提供了技术和特征数据的描述。