Xc10 digital camera

During cell culture, we confirmed the identity of tenocytes using the ICC with 2 × 10⁵ cells. After transferring 200 μL of cell culture to the 8-wells of a chamber slide, the cells were grown to confluence with the addition of fresh media for one day. After washing with PBS, the cells were fixed with 4% formaldehyde. Permeabilization of the membranes was done by incubating the slides with 0.25–0.5% Triton X-100 in PBS for 10 min, followed by blocking with rabbit normal serum, donkey normal serum, for vimentin and tenomodulin, respectively. The cells were incubated in the dark with mouse anti-vimentin monoclonal antibody (1:100 dilution; Dako, Glostrup, Denmark; code: M0725) and goat anti-tenomodulin antibody (1:25 dilution; Santa Cruz Biotechnology, Santa Cruz, CA, USA; code: sc-49325) for 60 min at 37 °C. After additional washing, the secondary antibodies were incubated in the dark for 30 min at 37 °C, and mounted in a Vectashield H-1000 mounting medium for fluorescence (Vector Laboratories, Burlingame, CA, USA). Analysis was done using an Olympus CKX41 microscope equipped with epifluorescence and an Olympus XC10 digital camera. To determine the expression of vimentin or tenomodulin in the cultured cells, we randomly sampled 40 views of cell staining from two groups (healthy tenocyte, tendinopathy tenocyte), with approximately 1000 cells each.

All the fluorescence images were taken using an optical Olympus IX51 microscope equipped with a 100-W Mercury light source (U-LH100HG, Olympus, Tokyo, Japan). For actin filaments, visualization U-MWIG2 filter (λexit = 530−550 nm, λemit = 590 nm) was used, and U-MNB2 one (λexit = 470−490 nm, λemit = 520 nm) was used to detect fluorescently labeled cell nuclei. Fluorescence images were recorded using the XC10 digital camera (Olympus). The maximum resolution of the captured image by this camera was 2080 ×  1544 px. All the images were recorded using CellSense Dimensions (Olympus) software with the objective 20 × (Universal Plan Fluorite, the magnification of 200).