Kreckel Heidi D, Samuels Fionna M D, Bonnart Remi, Volk Gayle M, Stich Dominik G, Levinger Nancy E
Department of Chemistry, Colorado State University, Fort Collins, CO 80523, USA.
USDA ARS National Laboratory for Genetic Resources Preservation, Fort Collins, CO 80521, USA.
Plants (Basel). 2023 Jun 8;12(12):2247. doi: 10.3390/plants12122247.
Cryopreservation has emerged as a low-maintenance, cost-effective solution for the long-term preservation of vegetatively propagated crops. Shoot tip cryopreservation often makes use of vitrification methods that employ highly concentrated mixtures of cryoprotecting agents; however, little is understood as to how these cryoprotecting agents protect cells and tissues from freezing. In this study, we use coherent anti-Stokes Raman scattering microscopy to directly visualize where dimethyl sulfoxide (DMSO) localizes within shoot tips. We find that DMSO fully penetrates the shoot tip tissue within 10 min of exposure. Variations in signal intensities across images suggest that DMSO may interact with cellular components, leading to its accumulation in specific regions.
冷冻保存已成为一种用于长期保存无性繁殖作物的低维护、经济高效的解决方案。茎尖冷冻保存通常采用玻璃化方法，该方法使用高浓度的冷冻保护剂混合物；然而，对于这些冷冻保护剂如何保护细胞和组织免受冷冻的了解却很少。在本研究中，我们使用相干反斯托克斯拉曼散射显微镜直接观察二甲基亚砜（DMSO）在茎尖内的定位。我们发现，DMSO在暴露10分钟内即可完全穿透茎尖组织。图像中信号强度的变化表明，DMSO可能与细胞成分相互作用，导致其在特定区域积累。
