rnascope 2.5 hd duplex detection kit (chromogenic)  (Advanced Cell Diagnostics Inc)

Journal: Acta Neuropathologica Communications

Article Title: A comprehensive spatiotemporal map of dystrophin isoform expression in the developing and adult human brain

doi: 10.1186/s40478-025-01996-z

Figure Lengend Snippet: Human DMD transcripts and RNA labelling dynamics at the 5′ and 3′ ends of the DMD gene protein. Isoforms derived from the DMD gene can be classified into full-, intermediate- and short-length according to their mass in kDa and are all expressed in the human nervous system. DMD exons are shown with colours matching the encoded dystrophin functional domains. ASP (alternative splicing products) are generated by alternative splicing and distinct polyA-addition signals ( a ). The DMD 5′ probe detects dp427c , dp427m and dp427p and the DMD 3′ probe detects all the mature DMD transcripts except dp40 ( b ). Images show the variable size of the RNAscope red punctate dots detected using DMD 5′ and DMD 3′ probes ( c ). Scale bars = 50µm

Article Snippet: Singleplex RNAscope assays (Advanced Cell Diagnostics Inc, Cat. No. 322360) relying on RNAscope DMD 5′ and DMD 3′ probes were performed according to the manufacturer’s protocol.

Techniques: Derivative Assay, Functional Assay, Alternative Splicing, Generated, RNAscope

Journal: Acta Neuropathologica Communications

Article Title: A comprehensive spatiotemporal map of dystrophin isoform expression in the developing and adult human brain

doi: 10.1186/s40478-025-01996-z

Figure Lengend Snippet: In situ hybridisation in the human brain. Heatmap representing the DMD 5′ and DMD 3′ RNAscope signal scores in each adult ROIs ( a ). Heatmap representing the singleplex Basescope P+ (% Positive cells/total cells) across different adult brain areas ( b ). Colour code: white: unprocessed; light green: low expression; green: moderate expression; orange/red: high expression; grey: undetermined; S1-4: subjects processed for each developmental stage

Article Snippet: Singleplex RNAscope assays (Advanced Cell Diagnostics Inc, Cat. No. 322360) relying on RNAscope DMD 5′ and DMD 3′ probes were performed according to the manufacturer’s protocol.

Techniques: In Situ, Hybridization, RNAscope, Expressing

Journal: Acta Neuropathologica Communications

Article Title: A comprehensive spatiotemporal map of dystrophin isoform expression in the developing and adult human brain

doi: 10.1186/s40478-025-01996-z

Figure Lengend Snippet: DMD transcript detection by RNAscope in embryonic human brain at Carnegie stage 17 (CS17). Top left image shows a low magnification of a CS17 embryo sagittal section. Individual plates show representative images of brain sections from the regions indicated in the CS17 embryo hybridised either with the DMD 5′ or DMD 3′ probe. Note extensive DMD 3′ signal in all brain regions, whereas the DMD5′ signal is either not detectable or detected only in a few cells, with the exception of the trigeminal ganglion where several positive cells are observed. As positive control, representative images of FFPE muscle sections hybridised either with the DMD 5′ or the DMD 3′ probe are shown; note extensive signal with both probes (red dots). Inserts show representative puncta at a higher magnification. Scale bars are: embryo = 1 mm; images on left column = 10 µm; telencephalon = 25 µm; all other images = 50 µm

Article Snippet: Singleplex RNAscope assays (Advanced Cell Diagnostics Inc, Cat. No. 322360) relying on RNAscope DMD 5′ and DMD 3′ probes were performed according to the manufacturer’s protocol.

Techniques: RNAscope, Positive Control

Journal: Acta Neuropathologica Communications

Article Title: A comprehensive spatiotemporal map of dystrophin isoform expression in the developing and adult human brain

doi: 10.1186/s40478-025-01996-z

Figure Lengend Snippet: DMD transcript detection by RNAscope in embryonic human brain at Carnegie stage 23 (CS23). The left image shows a low magnification of a CS23 embryonic brain coronal section highlighting main structures at this stage of development. Right panels show representative images of telencephalon, hippocampus and cerebellum hybridised either with the DMD 5′ or DMD 3′ probe. Note extensive DMD 3′ signal in all brain regions with representative puncta shown at higher magnification in the inserts; the DMD 5′ signal is very sparse (telencephalon and cerebellum) or absent (hippocampus). CP choroid plexus, H hippocampus, LV lateral ventricle, 4V 4th ventricle. Scale bars are: CS23 embryo = 2 mm telencephalon and cerebellum = 100 µm; hippocampus = 250 µm

Article Snippet: Singleplex RNAscope assays (Advanced Cell Diagnostics Inc, Cat. No. 322360) relying on RNAscope DMD 5′ and DMD 3′ probes were performed according to the manufacturer’s protocol.

Techniques: RNAscope

Journal: Acta Neuropathologica Communications

Article Title: A comprehensive spatiotemporal map of dystrophin isoform expression in the developing and adult human brain

doi: 10.1186/s40478-025-01996-z

Figure Lengend Snippet: DMD transcript detection by RNAscope in a 19 weeks post-conception (PCW19) foetal human brain. Left images show a low magnification of PCW19 hippocampus and cerebellum with localisation of regions shown on the left indicated. Individual plates on the right show representative sections of hippocampus and cerebellum hybridised either with the DMD 5′ or DMD 3′ probe. Signal from both probes is detected in the dentate granule cells, CA1 and subiculum of the hippocampus, as well as in all cerebellar layers: external granular layer (EGL), molecular layer (MOL), Purkinje cell layer (PCL) and granule cell layer (GCL). Some representative puncta are shown at higher magnification in the inserts. Scale bars are: low magnification hippocampus and cerebellar cortex = 1 mm and = 500 µm, respectively; all other images = 40 µm

Article Snippet: Singleplex RNAscope assays (Advanced Cell Diagnostics Inc, Cat. No. 322360) relying on RNAscope DMD 5′ and DMD 3′ probes were performed according to the manufacturer’s protocol.

Techniques: RNAscope

Journal: Acta Neuropathologica Communications

Article Title: A comprehensive spatiotemporal map of dystrophin isoform expression in the developing and adult human brain

doi: 10.1186/s40478-025-01996-z

Figure Lengend Snippet: Detection of DMD isoforms, GABAergic and glutamatergic neurons in developing and adult human brain areas by duplex RNAscope. Representative image of experiments performed in 14pcw cerebellum ( a – d ). Representative images of the results from the adult samples ( e – n ). Green dots indicate RNAscope DMD 5′ and DMD 3′ transcripts. Red signal indicates GAD1 (GABAergic) or SLC17A7 (glutamatergic) transcripts. Black arrows indicate migrating Purkinje neurons ( a ) external granular layer ( c ) GAD1 - or low GAD1 + Purkinje neurons ( e , g ). The green arrow shows sporadic DMD 3′ signal in d . Red arrows indicate GAD1 + Purkinje neurons ( e , g ) and GAD1 + dentate neurons ( f , h ) in the cerebellum. Scale bars are: a, e, l = 50 µm; all other images = 10 µm

Article Snippet: Singleplex RNAscope assays (Advanced Cell Diagnostics Inc, Cat. No. 322360) relying on RNAscope DMD 5′ and DMD 3′ probes were performed according to the manufacturer’s protocol.

Techniques: RNAscope

Journal: Acta Neuropathologica Communications

Article Title: A comprehensive spatiotemporal map of dystrophin isoform expression in the developing and adult human brain

doi: 10.1186/s40478-025-01996-z

Figure Lengend Snippet: Dystrophin detection by immunohistochemistry in developing and adult brain regions. Developing brains stained either with the monoclonal antibody DYSA (dystrophin N-Terminus) or DYS2 (dystrophin C-Terminus) ( a – c ). Adult brain sections stained with DYSA ( d ). Adult upper and lower motor neurons stained with DYSA and RNAscope DMD 5′ and DMD 3′ probes ( e – f ). Pcw postconceptional weeks, CS Carnegie stage. Scale bars are: a = 1 mm (overview) and 50 µm; d = 100 µm; b and c = 50 µm all other images = 10 µm

Article Snippet: Singleplex RNAscope assays (Advanced Cell Diagnostics Inc, Cat. No. 322360) relying on RNAscope DMD 5′ and DMD 3′ probes were performed according to the manufacturer’s protocol.

Techniques: Immunohistochemistry, Staining, RNAscope

Journal: Acta Neuropathologica Communications

Article Title: A comprehensive spatiotemporal map of dystrophin isoform expression in the developing and adult human brain

doi: 10.1186/s40478-025-01996-z

Figure Lengend Snippet: Dystrophin protein and transcript expression in brain vasculature. Embryonic and foetal choroid plexus and blood vessels stained with DYSA and DYS2 antibodies and hybridised with the RNAscope DMD 5′ and DMD 3′ probes ( a – e ). Adult blood vessels stained with DYSA antibody and processed for RNAscope or Duplex Basescope ( f – i ). Note positive signal detected with duplex Basescope ( Dp427m -green/ Dp71 -red), RNAscope DMD 5′ and DMD 3′ probes, and the DYSA antibody within the tunica media of a medium-sized leptomeningeal artery ( f ) and in a cortical penetrating arteriole ( g ) in the ventromedial prefrontal/orbitofrontal cortex. Note also expression of the Dp71 transcript and DYSA immunoreactivity in the cerebellar cortical parenchymal capillary ( h ) and cerebellar pial–glial membrane (glia limitans) at the cortical surface ( i ). Each red arrow indicates RNAscope or duplex Basescope ( dp71 ) signal (red dots) while green arrows indicate duplex Basescope ( dp427m ) signal (green dots). Scale bars are: f, g, h, i = 25 µm; all other images = 10 µm

Article Snippet: Singleplex RNAscope assays (Advanced Cell Diagnostics Inc, Cat. No. 322360) relying on RNAscope DMD 5′ and DMD 3′ probes were performed according to the manufacturer’s protocol.

Techniques: Expressing, Staining, RNAscope, Membrane