Position-referenced microscopy: Regions of interest localization and subpixel image comparison by means of pseudo-random patterns embedded in cell culture boxes

Abstract

This work presents micro-grids integrated to cell culture boxes. These grids allow the systematic registration of the position of a zone observed by optical microscopy in a such way that it is possible to find it again easily for new observations for instance after culture on drug injection. The position knowledge allows also the numerical superimposition of recorded images in a common position reference system with a sub-pixel precision. It become thus straightforward to perform a site by site analysis of the possible evolutions that may have occurred in the biological medium.

Position-referenced microscopy for live cell culture monitoring

“…3(a). The propagation of the altered spherical wave up to the focusing plane has been calculated by using the angular spectrum of plane waves approach in the scalar approximation [14, 15, 17]. A 2D Fourier transform is first applied to the converging beam in the plane just behind the PRP.…”

“…The method capabilities to provide absolute and high-accuracy in-plane coordinates of the fields of observation from the processing of the PRP images have been demonstrated elsewhere [13, 17, 18]. Resolutions achieved are in the range of a few nanometers for lateral position and of 10 −3 degree for in-plane orientation.…”