Paired bright-field and DAPI time-lapse microscopy dataset of fibroblast-like cells (synoviocytes) from osteoarthritic knee tissue.

Dataset accompanying the article: “A deterministic method for quantifying spindle-shaped cells in noisy bright-field microscopy.” This dataset contains paired microscopy images used for the evaluation of a deterministic, label-free method for quantifying spindle-shaped cells in noisy bright-field microscopy. In addition, the repository includes source code implementing a particle swarm optimization (PSO) procedure for tuning classification thresholds in the Gaussian-based contour analysis. The imaged cells are primary human fibroblast-like synoviocytes (FLS) derived from patient synovial tissue. Each field of view includes: a bright-field microscopy image a corresponding fluorescence image with DAPI-stained nuclei The fluorescence images were used exclusively for validation and quantitative performance assessment in the associated study. Bright-field Images Original bright-field microscopy images of primary human FLS characterized by: - low contrast - image noise - heterogeneous background - irregular spindle-shaped cell morphology These conditions reflect realistic experimental settings encountered in label-free time-lapse microscopy. DAPI Images Fluorescence microscopy images of the same fields of view with nuclei stained using DAPI. These images provide reference information for validation of detection and quantification results. Image Acquisition Parameters All time-lapse microscopy images were acquired using a Cytation 5 Cell Imaging Multi-Mode Reader (BioTek Instruments). Image acquisition was controlled using Gen5 software (version 3.11.19). Images were captured with a Blackfly BFLY-U3-23S6M monochrome camera (FLIR Systems) without binning. The camera operated with a 32 ms exposure time, gain set to 0.15, LED intensity level 4, brightness level 50, and contrast level 33. The saturation level of the sensor was 65504. Bright-field imaging was performed using a 4× Olympus objective (numerical aperture NA = 0.13). The effective image resolution was 1224 × 904 pixels, corresponding to a field of view of approximately 2017 µm × 1489 µm. Reduced field-of-view mode was enabled. Cells were maintained at 37 °C during imaging. No Z-stack acquisition was performed (single focal plane, ZStackTotal = 1). Time-lapse acquisition consisted of 200 kinetic reads per series with a temporal interval of 1,230,000 ms (approximately 20 minutes). Imaging was performed in 24-well plates (TPP, circular wells, diameter 20,000 µm). Fluorescence illumination correction was disabled. No phase contrast imaging was used. Bright-field imaging was performed in monochrome mode.