Lensless Digital Holographic Microscopy (LDHM) presents notable advantages, offering a wide field of view (FOV) for imaging with reasonable resolution, achieved through a relatively straightforward and cost-effective setup - typically comprising only a point source of light, a sample, and a camera. These properties, especially the wide FOV, make LDHM a promising tool for the population-wise characterization of biological cell cultures with single cell precision. Cell cultures grow in a controlled environment, and the slightest change in environmental conditions can have a significant impact. The same principles apply to imaging. Any alteration in the imaging process can disrupt natural growth and potentially lead to false conclusions during comparison to the control culture. A crucial challenge is to maintain the same environmental conditions. To address this, we propose a novel setup for simultaneous imaging on three parallel in-line LDHM systems, with cameras mounted on one rail (movable to realize multi-height hologram recording for iterative reconstruction), all sample holders mounted on the second rail, and all sources of light mounted on the third rail. Based on the collected holograms and their efficient reconstruction, it is possible to calculate migration parameters such as the average speed of a single cell or a predefined group of cells (e.g., big cells or slow cells) and statistics regarding cell number, size and speed over time.
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