Cells were filmed with a Axiocam MRm camera coupled to a Zeiss AxioImager A2 upright microscope fitted with a Zeiss 40×/0.75 Plan-Neofluar objective and X-Cite 120Q Metal Halide lamp, controlled by Axio Vision SE64 Rel 4.9.1. The resulting images were converted to black and white and analyzed using ImageJ 1.50i (Fig 1). Intensity of immunofluorescence was quantified by corrected total cell fluorescence (CTCF), by substracting background signal (average signal per pixel for a region selected just beside the cell) from whole cell signal (sum of the intensity of the pixels for one cell) [29 (link)]. For each sample, five CTCFs were measured and the average was calculated for statistical analysis.
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