Journal: PLoS ONE

Article Title: Analysis of Genome-Wide Monoallelic Expression Patterns in Three Major Cell Types of Mouse Visual Cortex Using Laser Capture Microdissection

doi: 10.1371/journal.pone.0163663

Figure Lengend Snippet: ( a ) Schematic flowchart of the experimental design. Top : mating strategy to obtain B6/CAST mice. Middle : 7 μm cryostat sections from P28 mice were cut and single cells were identified for laser capture microdissection (LCM) as excitatory neurons, inhibitory neurons or astrocytes in layer 2/3 of the visual cortex. Scale bar, 50 μm. Bottom : LCM single cells were collected and RNA was extracted. A cDNA library, was generated for each cell type followed by RNA-Seq, and allele-specific expression in the cellular transcriptome was analyzed. ( b ) Schematic diagram showing cross of cell-type-specific Cre lines with a Cre/loxP reporter line ( Ai14 ). ( c ) Confirmation of each specific cell type captured by LCM in Cre reporter cell lines. Left panels (i , v , ix) : Representative images of the visual cortex from the three cell-type-specific Cre mouse lines. Cells are visualized with tdTomato (red). Cell-type specific Cre reporter mouse lines are indicated above each image. Squares indicate areas enlarged in left panel. Scale bar, 100 μm. Right panels ( ii-iv , vi-viii , x-xii) : Enlarged images of cells labeled with antibodies (green) to anti-CAMK2A (excitatory neurons), anti-GAD2 (for inhibitory neurons) or anti-S100B (for astrocytes). Double-labeled cells (overlay) confirmed cell type of red fluorescent cells from the cell-type-specific Cre reporter lines. Sections were counterstained with DAPI (blue). Scale bar, 20 μm.

Article Snippet: Red fluorescent positive cells in layer 2/3 of primary visual cortex were captured using a laser capture microdissection system (Zeiss, PALM MicroBeam) and collected into AdhesiveCap 500 opaque PCR tubes (Zeiss, 415190-9201-000).

Techniques: Laser Capture Microdissection, cDNA Library Assay, Generated, RNA Sequencing, Expressing, Labeling

Journal: iScience

Article Title: Investigating microglia-neuron crosstalk by characterizing microglial contamination in human and mouse patch-seq datasets

doi: 10.1016/j.isci.2023.107329

Figure Lengend Snippet:

Article Snippet: Patch-Seq of neurons in mouse primary visual cortex , Gouwens et al., , Allen Institute for Brain Science Cell Types Database.

Techniques: Software