β tubulin mouse polyclonal antibody  (Proteintech)

Journal: Advanced Healthcare Materials

Article Title: Smart Polymeric 3D Microscaffolds Hosting Spheroids for Neuronal Research via Quantum Metrology

doi: 10.1002/adhm.202403875

Figure Lengend Snippet: Evaluation of embryoid bodies (EBs) interaction with the 3D scaffold. a) EBs seeded on a planar coverslip showing dopaminergic neurons‐specific tyrosine hydroxylase (TH) staining in magenta (AF 594) and cells nuclei in green (DAPI). b) EBs seeded on a collagen droplet showing an extensive neuronal network of beta‐III tubulin incyan (AF 488) and TH (magenta) neurites. c) EBs on spiderweb design showing cells growing around the scaffold, using it for support and guidance. d) The green channel in the image shows cell nuclei and scaffold structure. e) A robust network of β ‐III tubulin‐positive neurites extending from the EB center and integrating with the scaffold. f) TH‐positive cells visible around the EB, predominantly surrounding the scaffold. g) Channels merge and overview of the EB and 3D scaffold interaction. h) Close‐up view from g) highlighting the deep interactions between neurites and the scaffold, with cells surrounding and stretching along it. i) Higher magnification of cells extending over the scaffold beams and expressing β ‐III tubulin and TH, confirming scaffold‐cell interactions. j) The Scanning Electron Microscopy (SEM) image shows significant neurite interaction with the scaffold. Green channel: DAPI ‐nucleus/scaffold. Cyan channel: β ‐III tubulin marker – neurons. Magenta channel: TH marker – dopaminergic neurons. The scale bar represents 50 µm.

Article Snippet: EBs were then incubated overnight at 4 °C with a single pair of primary antibodies, mouse polyclonal β ‐III tubulin antibody (1:1000; Promega) and rabbit polyclonal TH antibody (1:500; Abcam), diluted in blocking buffer.

Techniques: Staining, Expressing, Electron Microscopy, Marker

Journal: Advanced Healthcare Materials

Article Title: Smart Polymeric 3D Microscaffolds Hosting Spheroids for Neuronal Research via Quantum Metrology

doi: 10.1002/adhm.202403875

Figure Lengend Snippet: Orientation distribution analysis of neuron alignment with the scaffold. a) Two‐color confocal image of an embryoid body (EB) in interaction with 3D Block Spiderweb microscaffold. b) Distribution of neuron orientations in degrees, comparing different gradient methods: Cubic spline, Fourier, Gaussian, and Hessian. All methods produced similar results with similar peak positions for scaffold and cell orientations. c) Three‐color confocal image of an EB in interaction with 3D Block Spiderweb microscaffold. d) The orientation distribution profile is derived from the image. c) The alignment of dopaminergic neurons (red) and the other neurons (green). Dopaminergic neurons align with both −45 °and 45 °peaks, while other neurons here primarily align with −45°. The analysis confirms that neurons consistently tend to align with scaffold orientation peaks, validating the scaffold's effectiveness in influencing the growth of 3D culture models. Blue channel: scaffold. Green channel and red channel in a): β ‐III marker – neurons. Red channel in c): Tyrosine Hydroxylase (TH) marker – dopaminergic neurons. The scale bar represents 50 µm.

Article Snippet: EBs were then incubated overnight at 4 °C with a single pair of primary antibodies, mouse polyclonal β ‐III tubulin antibody (1:1000; Promega) and rabbit polyclonal TH antibody (1:500; Abcam), diluted in blocking buffer.

Techniques: Blocking Assay, Produced, Derivative Assay, Marker