Journal: Matrix biology : journal of the International Society for Matrix Biology

Article Title: Collagen fiber structure guides 3D motility of cytotoxic T lymphocytes

doi: 10.1016/j.matbio.2019.02.003

Figure Lengend Snippet: Alignment of collagen fibers in microfluidic devices. A. Schematic of microfluidic device for fiber alignment. Channel widths are 250 × 250 μm for the small channel and 250 μm × 2 mm for the large channel. Drawing to scale. B. Representative reflective confocal image of collagen ultrastructure in large and small channels, scale bar = 100 μm. (C) MatFiber output with arrows tracing collagen fibers to indicate directionality in unaligned and aligned channels. Scale bar = 200 μm. (D) Probability distribution of angles of collagen fibers in aligned and unaligned conditions. These experiments were independently repeated three times.

Article Snippet: Collagen fiber density and orientation analysis was adapted from a previously developed method using MATLAB (Mathworks) software MatFiber [ 17 ].

Techniques:

Journal: Matrix biology : journal of the International Society for Matrix Biology

Article Title: Collagen fiber structure guides 3D motility of cytotoxic T lymphocytes

doi: 10.1016/j.matbio.2019.02.003

Figure Lengend Snippet: CD8+T cells colocalize with aligned collagen I fibers in tumors. (A) Representative SHG image of TRAMPC2 xenograft tumors. (B) Matfiber alignment vectors of SHG image. (C) Histogram of collagen alignment profiles in TRAMPC2 tumors (n = 8 tumors) Xenograft tumors (n = 4) stained for collagen I (green) and CD8 (red; nuclei in blue): (D) Left-Stromal regions adjacent to TRAMPC2 subcutaneous xenograft tumors and right- high magnification of the boxed area on the right, showing CD8+ T cells localized between collagen fibers. (E) Right - periphery of TRAMPC2 tumors depicting infiltrating CD8+ T cells and right- high magnification of the boxed area on the right, showing CD8+ T cell located along aligned collagen fibers. Scale bars = 50 μm. (F) Photomicrograph depicting T cells crawling along collagen fibers using SHG imaging (G) Schematic depiction of method for quantifying T cell association with collagen fibers. (H) Scatter plot of collagen fiber and T cell angles (n = 26 cells) showing positive correlation. (I) CD8 immunostaining of central regions and peripheral regions of TRAMPC2 tumors. (J) Quantification of CD8+ cells within different regions of the tumor. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Collagen fiber density and orientation analysis was adapted from a previously developed method using MATLAB (Mathworks) software MatFiber [ 17 ].

Techniques: Staining, Imaging, Immunostaining