Scholl Cheyanne M, Li Sophia, Flanders Wyatt, Basran Parminder S
Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.
Med Phys. 2025 Aug;52(8):e18004. doi: 10.1002/mp.18004.
A central challenge in classical radiobiology experiments, where dishes are plated with cells and then irradiated, is that the radiation dose deposited to the cells is often subjected to systematic and random errors. One method to validate the dose to cells is to use radiochromic film underneath or above the plated cells to estimate the dose given to the culture.
To explore the feasibility of seeding cells directly onto radiochromic film, enabling precise characterization of cellular responses to microscopic fluctuations in radiation intensity. This technique decreases the challenge of registration, improving the correlation between individual cell and their received radiation dose.
We investigate several adhesives and strategies for adhering spindle cells on a thin layer atop the film. After finding a robust approach, we develop a novel strategy for absolute dose calibration to the cell-film substrates. Finally, we evaluate this approach using a standard radiobiology assay by exposing cell films to uniform doses and comparing the cell colony survival fraction with established and published data.
Easily obtainable gelatin and 3D printing adhesives provide substrates and adhesives for cell colony formation. Calibration and sample film data from traditional flatbed scanners used for film dosimetry, confocal microscopes, and a novel calibration method can be used to measure the dose to the films.
Using a film-to-dose calibration methodology, we show that cell colony assay experiments can be conducted directly on radiochromic film without significant spatial variation.
在经典放射生物学实验中,将细胞接种在培养皿中然后进行照射,一个核心挑战是沉积到细胞上的辐射剂量常常受到系统误差和随机误差的影响。一种验证细胞剂量的方法是在接种的细胞下方或上方使用放射变色薄膜来估计给予培养物的剂量。
探索将细胞直接接种到放射变色薄膜上的可行性,从而能够精确表征细胞对辐射强度微观波动的反应。该技术降低了配准的难度,提高了单个细胞与其接受的辐射剂量之间的相关性。
我们研究了几种用于将纺锤状细胞粘附在薄膜上方薄层上的粘合剂和策略。找到一种可靠的方法后,我们开发了一种针对细胞 - 薄膜底物的绝对剂量校准新策略。最后,我们通过将细胞薄膜暴露于均匀剂量并将细胞集落存活分数与已建立和已发表的数据进行比较,使用标准放射生物学测定法评估这种方法。
易于获得的明胶和3D打印粘合剂为细胞集落形成提供了底物和粘合剂。来自用于薄膜剂量测定的传统平板扫描仪、共聚焦显微镜的校准和样品薄膜数据以及一种新的校准方法可用于测量薄膜的剂量。
使用薄膜到剂量的校准方法,我们表明细胞集落测定实验可以直接在放射变色薄膜上进行,且无明显空间变化。