A Petri-Dish with Micromolded Pattern as a Coordinate Indicator for Live-Cell Time Lapse Microscopy.

Determining the position of cells in a microscope image is essential in investigating cell behaviors such as migration and differentiation. When observing cells over a time period over many ROIs (region of interests), it is difficult to reposition the sample and find the same cell once the FOV (field of view) is changed. This study describes a novel approach to obtain accurate position data in live-cell time-lapse imaging and large-area stitched images using a Petri-dish with embedded microstructures and a simple microscope. We integrated a braille-like pattern underneath the Petri-dish, a coordinate indicator for microscope image registration to simplify imaging of multiple ROIs at different time points without expensive equipment such as motorized stages or enclosed cell culture incubators. In this study, we micromolded microscopic structures with sizes between 5–10 μm spaced closely (~ 40 μm) in non-repeating pattern underneath the petri-dish. Image processing technique was used to determine the position and rotation of the petri-dish with respect to the microscope FOV. Using this methods, we demonstrated successful imaging of time-lapse images for cell differentiation and migration. Large area patterned cell cultures were imaged over several days and the images were stitched without a motorized microscope. [ABSTRACT FROM AUTHOR]