Snapshots in Neuroscience: Multiplex RNA imaging

These images have been selected to showcase the art that neuroscience research can create.

As described by the authors: “Shown is a 63X tile scan image from a juvenile mouse hippocampus stained using highly multiplexed single molecule fluorescent in situ hybridization (HiPlex smFISH). Twelve mRNAs were simultaneously hybridized and then iteratively labeled and imaged in three separate rounds. The image shows mRNA expression for four mRNAs (Adcy1, Aco2, Psd, Dlg4) after the first round of imaging. These mRNAs are abundantly expressed in the neuropil, but also differentially enriched in specific hippocampal subregions (CA1, CA2, CA3, DG).

“In our study, HiPlex smFISH was used to investigate the spatial organization of localized mRNAs in the mouse hippocampus. The laminar cytoarchitecture of the hippocampus allowed us to visualize and spatially map mRNAs localized to the synapse-rich neuropil layer. We gained insights into how the spatial distribution and colocalization of these mRNAs are, in large part, explained by stochastic interactions based on abundance rather than coordinated co-packaging of these specific mRNAs into defined granules.

“The image was acquired on a Leica DMi8 inverted Thunder widefield microscope using a 63X/1.4 NA objective and postprocessed using Thunder large volume computational clearing. Adcy1 (green), Aco2 (magenta), Psd (cyan), and Dlg4 (yellow) mRNAs were labeled using HiPlex RNAscope in situ hybridization assay. DAPI was used to counterstain nuclei (blue).”

A 63X tile scan image of four neuropil-localized mRNAs in a horizontal section from a juvenile mouse hippocampus. Adcy1 (green), Aco2 (magenta), Psd (cyan), and Dlg4 (yellow) mRNAs were labeled using HiPlex RNAscope in situ hybridization. DAPI was used to counterstain cell nuclei (blue). Image credit: Shannon Farris.

Read the full article:

Multiplexed smFISH Reveals the Spatial Organization of Neuropil Localized mRNAs Is Linked to Abundance
Renesa Tarannum, Grace Mun, Fatima Quddos, Sharon A. Swanger, Oswald Steward, and Shannon Farris