The lateral ventricle walls contain the largest germinal region in the adult mammalian brain. Traditionally, studies on neurogenesis in this region have relied on classical sectioning techniques for histological analysis. Here we present an alternative approach, the wholemount technique, which provides a comprehensive, en-face view of this germinal region.
Total time
~3–4 hours per brain (dissection, flow analysis, and staining combined)
Model organism
Mouse (adult, strain not specified)
Steps
1
Introduce subventricular zone anatomy and wholemount advantages
Overview of the lateral ventricle as the largest adult mammalian germinal region and explanation of why en-face wholemount preparation offers superior comprehensive visualization compared to classical sectioning.
▶ 00:07
2
Dissect and prepare lateral ventricle wholemounts
Detailed protocol for removing and carefully unfolding the lateral ventricle walls to create a flat, intact tissue preparation suitable for en-face imaging and downstream analysis.
▶ 01:02
3
Track ependymal flow with fluorescent microbeads
Application of fluorescent microbeads to the ependymal surface to visualize and measure the directional flow characteristics of cilia-driven fluid movement across the germinal zone.
▶ 04:14
4
Fix and immunostain wholemount tissue preparations
Protocol for chemical fixation and multi-target immunofluorescence staining of intact wholemounts to label neurogenic markers and cellular populations within the subventricular zone.
▶ 06:41
5
Interpret representative images and flow quantification
Analysis and visualization of typical en-face wholemount images and quantitative ependymal flow data, demonstrating output quality and biological interpretation.
▶ 10:58