Journal: bioRxiv
Article Title: RatDISCO, a tissue clearing and immunolabelling protocol for large rat brains
doi: 10.1101/2025.08.17.670723
Figure Lengend Snippet: RatDISCO enables immunodetection of neuronal markers, viral tracing, and transgenic expression in mice and human brain organoids. Light-sheet microscopy images of whole-mouse brains immunolabelled for endogenous neuronal markers: MECP2 (A), cFOS (B, visualized with Inferno LUT), tyrosine hydroxylase (C), and FoxP2 (G: whole brain MIP; H-Optical slices at 0.84mm lateral to Bregma). D-F. Immunolabelling against virally-labelled medial (MEC) and lateral entorhinal cortex (LEC) neurons and axons in a Sim1 Cre mouse. (D) Whole-brain view of MEC projections. (E) Higher-magnification view of MEC-hippocampal projections. (F) Whole-brain view and a close-up of virally labelled MEC (GFP, green), LEC (mCherry, red) projections and merged image. (I) Immunolabelling of GFP in the transgenic P038 mouse line, highlighting MEC expression. Insets show GFP (green) co-labelling with parvalbumin (red) and the merged signal in a horizontal view. (J–O) Human iPSC-derived brain organoids (J, M; TO-PRO, magenta) co-cultured with GFP-positive single tumour cells (K, cyan) or tumour spheroids (N, cyan), with merged images shown in (L, O). Scale bars A-D, G-H: 500 µm. E: 250 µm, F, J-O and I-insets: 100 µm.
Article Snippet: Sim1 Cre mice (generated by GenSat and obtained from MMRRC: Tg(Sim1cre) KH21Gsat/Mmucd) were bred to be heterozygous for the Cre transgene by crossing male mice carrying the transgene with female C57BL6/J mice (Charles River, UK).
Techniques: Immunodetection, Transgenic Assay, Expressing, Microscopy, Derivative Assay, Cell Culture