instant structured illumination microscopy isim microscope  (Visitech Inc)

Journal: Mucosal Immunology

Article Title: γδ T cells compose a developmentally regulated intrauterine population and protect against vaginal candidiasis

doi: 10.1038/s41385-020-0305-7

Figure Lengend Snippet: a Left: Flow cytometry of CD3 + lymphocytes from the uterus of 2- and 12-week-old C57BL/6J mice. Representative plots are shown. Right: Uterine T-cell kinetics; the percentages of TCRδ + and TCRβ + cells (out of CD3 + cells) are indicated ( n = 3–7 per condition; a pool of nine uteri is shown for 1-week-old mice). Data represent at least two independent experiments. Graph indicates mean ± SD. b CD3 and TCRδ staining were analysed by confocal microscopy on uterus whole-mounts of C57BL/6J mice of the indicated ages. Representative images from two independent experiments are shown ( n = 3). Scale bars: 50 μm. c High-resolution images of CD3 and TCRγδ staining were obtained by iSIM on uterus whole-mounts of 2-week-old C57BL/6J mice ( n = 3). Scale bars: 10 μm. CD3-TCRδ co-staining appears as purple-white. d CD3 and EpCAM staining were analysed by immunohistochemistry on FFPE uterine serial sections. Arrows indicate the position of several CD3 + cells. Representative images from four independent samples are shown. e CD3 and EpCAM staining were analysed by confocal microscopy on uterus FFPE sections. Representative images are shown.

Article Snippet: For high-resolution analysis, samples were scanned using an Instant Structured Illumination Microscope (VT-iSIM, VisiTech International) using a ×100 1.45 NA objective and analysed with Nikon or Fiji (NIH).

Techniques: Flow Cytometry, Staining, Confocal Microscopy, Immunohistochemistry