Visual in-plane positioning of a Labeled target with subpixel Resolution: basics and application

Patrick Sandoz, July A. Galeano Zea, Artur Zarzycki, Deivid J. Botina Monsalve, Fabián Cortés Mancera, Andrés Cardona Echeverry, Laurent Robert


Vision is a convenient tool for position measurements. In this paper, we present several applications in which a reference pattern can be defined on the target for a priori knowledge of image features and further optimization by software. Selecting pseudo-periodic patterns leads to high resolution in absolute phase measurements. This method is adapted to position encoding of live cell culture boxes. Our goal is to capture each biological image along with its absolute highly accurate position regarding the culture box itself. Thus, it becomes straightforward to find again an already observed region of interest when a culture box is brought back to the microscope stage from the cell incubator where it was temporarily placed for cell culture. In order to evaluate the performance of this method, we tested it during a wound healing assay of human liver tumor-derived cells. In this case, the procedure enabled more accurate measurements of the wound healing rate than the usual method. It was also applied to the characterization of the in-plane vibration amplitude from a tapered probe of a shear force microscope. The amplitude was interpolated by a quartz tuning fork with an attached pseudo-periodic pattern. Nanometer vibration amplitude resolution is achieved by processing the pattern images. Such pictures were recorded by using a common 20x magnification lens.


Fourier transform phase processing, visual in-plane position measurement, vibration amplitude, shear force microscopy, position referenced microscopy.

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