Imunohistochemical labelling of whole mount hippocampal slice cultures

Figure 1e:
Problems associated with conventional immunohistochemical labelling of whole mount hippocampal slice cultures. Slices cultured for 12 days were labelled with isolectin-b4 using conventional techniques and Z-series scans were taken using a confocal microscope. Slices were mounted using coverslip bridges to avoid squashing. Representative images of a whole mount slice are shown. Although labelling is clear near the top and bottom, there appears to be an unlabelled ‘blank space’ in the middle of the slice. The ‘blank space’ cannot be identified in serial coronal cryostat sections. The section shows the CA1 pyramidal cell layer. Red is MAP2, green connexin-43 and blue GFAP. Scale bar: 50 μm.

From: Improved Immunohistochemical Protocol for Organotypic Brain Slice Cultures.
Jinny J. Yoon,¹ Vithika Suri,² Louise F. B. Nicholson² and Colin R. Green ¹
1. Department of Ophthalmology, and 2. Department of Anatomy with Radiology and the Centre for Brain Research, University of Auckland, New Zealand.
See: Microscopy and Analysis 25(1):5-7 (AM), 2011