Image-Based In Situ Calibration Of An X-Ray Microtomosynthesis Scanner Of Histopathology Samples

Histopathology procedures often produce a large number of thin sections from each tissue sample. It can be time intensive to analyze each section. We previously developed an x-ray linear micro tomosynthesis method that produced depth-resolved image stacks of intact samples in a short time window, at 10 Вµm or higher resolution and sufficient soft-tissue contrast to guide the subsequent sectioning and analysis procedure.[1] However, we found that the image quality could be improved by calibrating the sample scan trajectory for each scan, to compensate for mechanical instabilities that varied from scan to scan. We present a method for in situ calibration based on attaching a layer of microbead markers to the sample stage, which does not interfere with the sample imaging.