# Time-lapse imaging of cells in spatially fractionated X-ray fields using a mini beam as an alternative to accelerator-based sub-millimeter beams

## TL;DR

Researchers developed a mini X-ray beam setup to study cell responses to spatially fractionated radiation, using fluorescence markers to track cell cycle changes.

## Contribution

A Pb sub-milli-collimator is proposed as a practical alternative to accelerator-based beams for radiobiological studies.

## Key findings

- HeLa cells showed G2/M phase arrest in irradiated regions under spatially fractionated X-ray fields.
- FUCCI markers effectively distinguished irradiated and non-irradiated cells at sub-millimeter scales.
- Pb collimators offer a versatile and accessible alternative to synchrotron-based beams for preliminary radiobiology experiments.

## Abstract

Due to the limited number of accelerator-based X-ray facilities worldwide that provide beams with an adjustable size, their application for radiobiological research purposes has been restricted. Thus, the development of alternative methods is of technical importance for investigating cell/tissue responses in spatially non-uniform radiation fields. In this study, we performed mini beam irradiation of cells using a lead (Pb) sub-milli-collimator as an alternative method to sub-millimeter beams. Also, we employed human cervical carcinoma HeLa cells and hTERT-immortalized fibroblast BJ-1 cells that express fluorescence ubiquitination-based cell-cycle indicators (FUCCI). Time-lapse imaging revealed differences in the behavior of HeLa and BJ-1 cells in spatially heterogeneous radiation fields; in the case of HeLa cells, G2/M phase-arrested cells in the cell population were clearly observed, distinguishing irradiated from non-irradiated cells at the sub-millimeter scale level. Our findings indicate that FUCCI can be useful as a biological dose indicator, depending on cell type, and Pb sub-milli-collimators show potential as a possible alternative to accelerator-based X-ray sub-millimeter beams for radiobiological research. The use of the collimators, unlike beamtime experiments in synchrotron facilities with the approval of the committee, is highly versatile and may be beneficial in preliminary studies in a normal laboratory environment.