Journal: Advanced Science
Article Title: In Vivo Cytosolic Delivery of Biomolecules into Neurons for Super‐Resolution Imaging and Genome Modification
doi: 10.1002/advs.202501033
Figure Lengend Snippet: Selective neuronal uptake of N1 peptide after systemic administration coupled with FUS‐mediated BBB opening, and intrathecal administration into the spinal cord. A) Schematic timeline illustrating the process of FUS‐mediated BBB opening facilitated by microbubbles, systemic administration of FITC‐N1, and its subsequent specific delivery to the brain parenchyma. B) Mouse brain atlas indicating the trajectory of ultrasound waves from the transducer (blue lines) and the targeted area (cyan ellipse). C) Large field‐of‐view image showing the area labeled with FITC‐N1 following FUS‐mediated BBB opening (FUS‐treated), with the contralateral side (without FUS treatment) serving as the control. D) Zoomed views of the FUS‐treated area and the contralateral control side. E) High‐magnification images (maximum intensity projection) illustrating the neuronal specificity of FITC‐N1 after FUS‐facilitated BBB opening, confirmed by colocalization with the neuronal marker NeuN. F) The percentage of FITC‐N1 + cells colocalized with NeuN + cells relative to the total number of FITC‐N1 + cells. A total of 776 FITC‐N1 + cells were analyzed and data are presented as mean ± s.d., n = 4 animals. G) Schematic representation of intrathecal injection of FITC‐N1 between the lumbar 3 (L3) and lumbar 5 (L5) vertebral levels. H) Upon intrathecal injection, FITC‐N1 diffuses within the cerebrospinal fluid (CSF), leading to its distribution in the brain and penetration into the spinal cord. I) Image of a sagittal spinal cord section labeled with FITC‐N1 after intrathecal injection. J,K) Images (maximum intensity projection) showing the neuronal specificity of FITC‐N1 in the spinal cord after intrathecal injection, confirmed by colocalization of FITC‐N1 + cells and NeuroTrace™ 640/660 + cells. J) was acquired from the gray matter (ROI1) and K) from the white matter (ROI2) in (I). White boxes in (I) do not indicate the actual size of images (J) and (K). L) Quantification of the percentage of FITC‐N1 + cells colocalized with NeuroTrace™ 640/660 + cells relative to the total number of FITC‐N1 + cells. A total of 753 FITC‐N1 + cells were analyzed. Data are presented as mean ± s.d., n = 4 animals. Scale bars: C) 1 mm; I) 0.5 mm; D) 200 µm; E,J,K) 50 µm.
Article Snippet: Whole brain section images were acquired using a VS120 virtual slide microscope (Olympus) equipped with a SOLA‐SE‐II solid‐state white light excitation source.
Techniques: Labeling, Control, Marker, Injection