Jin Qiaoling, Vogt Stefan, Lai Barry, Chen Si, Finney Lydia, Gleber Sophie-Charlotte, Ward Jesse, Deng Junjing, Mak Rachel, Moonier Nena, Jacobsen Chris
Department of Physics & Astronomy, Northwestern University, Evanston, Illinois, USA.
X-ray Science Division, Advanced Photon Source, Argonne National Laboratory, Argonne, Illinois, USA.
PLoS One. 2015 Feb 23;10(2):e0117437. doi: 10.1371/journal.pone.0117437. eCollection 2015.
Rapidly-frozen hydrated (cryopreserved) specimens combined with cryo-scanning x-ray fluorescence microscopy provide an ideal approach for investigating elemental distributions in biological cells and tissues. However, because cryopreservation does not deactivate potentially infectious agents associated with Risk Group 2 biological materials, one must be concerned with contamination of expensive and complicated cryogenic x-ray microscopes when working with such materials. We employed ultraviolet germicidal irradiation to decontaminate previously cryopreserved cells under liquid nitrogen, and then investigated its effects on elemental distributions under both frozen hydrated and freeze dried states with x-ray fluorescence microscopy. We show that the contents and distributions of most biologically important elements remain nearly unchanged when compared with non-ultraviolet-irradiated counterparts, even after multiple cycles of ultraviolet germicidal irradiation and cryogenic x-ray imaging. This provides a potential pathway for rendering Risk Group 2 biological materials safe for handling in multiuser cryogenic x-ray microscopes without affecting the fidelity of the results.
快速冷冻的水合(冷冻保存)标本与低温扫描X射线荧光显微镜相结合,为研究生物细胞和组织中的元素分布提供了一种理想的方法。然而,由于冷冻保存不会使与风险组2生物材料相关的潜在感染因子失活,因此在处理此类材料时,必须关注昂贵且复杂的低温X射线显微镜受到污染的问题。我们采用紫外线杀菌照射对液氮下预先冷冻保存的细胞进行去污,然后用X射线荧光显微镜研究其在冷冻水合和冷冻干燥状态下对元素分布的影响。我们发现,与未经过紫外线照射的对照相比,即使经过多次紫外线杀菌照射和低温X射线成像循环,大多数对生物学重要的元素的含量和分布仍几乎保持不变。这为使风险组2生物材料在多用户低温X射线显微镜中安全处理提供了一条潜在途径,而不会影响结果的保真度。