acd rnascope ls multiplex fluorescent reagent kit  (Advanced Cell Diagnostics Inc)

Journal: Molecular Therapy. Methods & Clinical Development

Article Title: Serum factors create species-specific barriers to hepatic gene transfer by lipid nanoparticles in liver-humanized mice

doi: 10.1016/j.omtm.2025.101470

Figure Lengend Snippet: Biodistribution of DasherGFP mRNA in liver-humanized NSG-PiZ and FRG mouse liver RNAscope was performed on sections of livers from chimeric NSG-PiZ (A) or FRG (B and C) mice at 24 h post treatment with PBS or LNP1, LNP2, or LNP3 formulated with DasherGFP mRNA. High-power images of mouse hepatocytes from LNP2-treated (white arrows) but not LNP3-treated (red arrows) FRG mice demonstrate diffuse intracellular DasherGFP mRNA signal (C). Individual mouse IDs are indicated on the left. Sections were hybridized with probes specific for DasherGFP (green), human B2M (purple), and mouse GAPDH (cyan) and stained with DAPI (blue, A and B; white, C). Scale bars, 50 μm.

Article Snippet: In situ hybridization was performed using the ACD RNAscope LS Multiplex Fluorescent Reagent Kit (Advanced Cell Diagnostics, Newark, CA) and detection was performed using Akoya’s Opal Reagents (Akoya Biosciences).

Techniques: RNAscope, Staining

Journal: Molecular Neurodegeneration

Article Title: Amyloid-β (Aβ) immunotherapy induced microhemorrhages are linked to vascular inflammation and cerebrovascular damage in a mouse model of Alzheimer’s disease

doi: 10.1186/s13024-024-00758-0

Figure Lengend Snippet: Amyloid immunotherapy is highly associated with increased Trem2 + macrophages in 3D6 treated PDAPP Mice. a Dual RNAscope in situ hybridization of tissue-resident macrophages (Mrc1, green), monocyte-derived macrophages (CD74, purple) and immunofluorescence of amyloid (yellow) across the leptomeninges (colored lines) in PDAPP mice treated with 3D6 or IgG control. Amyloid, Mrc1 and CD74 overlay (Merge). b Quantification of leptomeningeal Mrc1 + area (%) of IgG or 3D6 treated PDAPP mice. c Quantification of leptomeningeal CD74 + area (%) in IgG or 3D6 treated PDAPP mice. d Dual RNAscope in situ hybridization of tissue-resident macrophages (Mrc1, green), monocyte-derived macrophages (CD74, purple) and immunofluorescence of amyloid (yellow) around amyloid + vessels in PDAPP mice treated with 3D6 or IgG control. Amyloid, Mrc1 and CD74 overlay (Merge). e Quantification of Mrc1 + area (%) of amyloid + vessels in IgG or 3D6 treated PDAPP mice. f Quantification of CD74 + area (%) of amyloid + vessels in IgG or 3D6 treated PDAPP mice. g Four-color immunofluorescences of amyloid (X-34, blue), endothelial cells (Pecam-1, cyan), tissue-resident macrophages (CD36, green, white arrow) and monocyte-derived macrophages (MHC II, red, yellow arrow) around vascular amyloid deposits in PDAPP mice treated with 3D6. h Multiplex RNAscope image of Trem2 (red), tissue-resident macrophages (Mrc1, green) and monocyte-derived macrophages (CD74, purple) around vessels in PDAPP mice treated with 3D6 or IgG control. Trem2, Mrc1 and CD74 immunoreactivity overlay (Merge). i Quantification of Trem2 + area (%) of vessels in IgG or 3D6 treated PDAPP mice. The number of vessels analyzed was 20. j Triple immunofluorescence of amyloid + (yellow) or amyloid − vessels, Trem2 (red) and dapi in PDAPP mice treated with IgG control. Amyloid, Dapi, Trem2 immunoreactivity overlay (Merge). k Quantification of Trem2 + area (%) of IgG in amyloid + or amyloid − vessels. l Triple immunofluorescence of amyloid + (yellow) or amyloid − vessels, Trem2 (red) and dapi in PDAPP mice treated with 3D6. m Quantification of Trem2 + area (%) of 3D6 in amyloid + or amyloid − vessels. The number of vessels analyzed was 3–7 per animal. Results are shown as mean ± SEM of n = 6 (mice). Asterisks indicate significant differences, where * p < 0.05, by unpaired Student’s t test. Scale bar 50 μm, 10 μm, 5 μm or 2 μm respectively

Article Snippet: Incubation, wash, and probe application periods were performed by utilizing the RNAscope® LS Multiplex Fluorescent Reagent kit system (Advanced Cell Diagnostics) and RNAscope® LS 4-plex Ancillary Kit for Multiplex Fluorescent Reagent (Advanced Cell Diagnostics) on the BOND RX (Leica).

Techniques: RNAscope, In Situ Hybridization, Derivative Assay, Immunofluorescence, Control, Multiplex Assay