Journal: bioRxiv
Article Title: Infection-on-Chip: an in vitro human vessel to study Neisseria meningitidis colonization and vascular damages
doi: 10.1101/2024.02.09.579276
Figure Lengend Snippet: Mimicking the in vivo vascular geometry using photoablation (A) Schematic representation of the development of the VoC: (i) a collagen-based hydrogel is inserted at the center channel of the microfluidic device, (ii) the focused UV-LASER locally carves the chosen geometry within the collagen I matrix, (iii) HUVEC attach on the collagen-carved scaffold. (B) Schematic representation of the microfluidic device and zoom in of the carved region after endothelialization (F-actin). (C) Schematic representation of the animal models assessed in this study: the mouse ear and the human skin-grafted mice. (D) Confocal image of vessel formation on 2.4 mg/ml - 3.5 mg/ml - 4 mg/ml and 6 mg/ml collagen I. The Corning collagen I has been used making the solution from 3.5 mg/ml to 6 mg/ml, and the Fujifilm collagen I has been used to make the 2.4 mg/ml. Each image is representative of one of the highest values of sprouting per condition. (E) Graph representing the number of sprouts per vessel for four collagen I concentrations: 2.4 mg/ml (FujiFilm), 3.5 mg/ml, 4 mg/ml and 6 mg/ml (Corning). Each dot represents one vessel. For each condition, the mean ± s.d. are represented (2.4 mg/ml: 5.20 ± 6.0 - 3.5 mg/ml: 1.69 ± 3.04 – 4 mg/ml: 0.045 ± 0.22 – 6 mg/ml: 0.23 ± 0.43). (F) Confocal images of F-actin and related Z-slice of the tissue-engineering VoC. (G) The photoablation technique to create in vivo -like structures: (i) definition of the region of interest to replicate from intravital microscopy images, (ii) image binarization, image post-processing and hydrogel-carving process, (iii) final results: immunostaining of the microfluidic chip.
Article Snippet: FujiFilm collagen I (LabChem Wako, Collagen-Gel Culturing Kit) has been used to obtain the 2.4 mg/ml solution, while the high concentration collagen I (Corning, #354249) has been used to obtain the 6 mg/ml, 4 mg/ml, and 3.5 mg/ml solutions according to the manufacturer’s protocol.
Techniques: In Vivo, Intravital Microscopy, Immunostaining
