Fumoto Shintaro, Kinoshita Eriko, Ohta Keisuke, Nakamura Kei-Ichiro, Hirayama Tasuku, Nagasawa Hideko, Hu Die, Okami Kazuya, Kato Riku, Shimokawa Shojiro, Ohira Naho, Nishimura Koyo, Miyamoto Hirotaka, Tanaka Takashi, Kawakami Shigeru, Nishida Koyo
Graduate School of Biomedical Sciences, Nagasaki University, Nagasaki 852-8501, Japan.
Division of Microscopic and Developmental Anatomy, Department of Anatomy, Kurume University School of Medicine, Kurume 830-0011, Japan.
Pharmaceutics. 2020 Nov 9;12(11):1070. doi: 10.3390/pharmaceutics12111070.
Visualizing biological events and states to resolve biological questions is challenging. Tissue clearing permits three-dimensional multicolor imaging. Here, we describe a pH-adjustable tissue clearing solution, Seebest (SEE Biological Events and States in Tissues), which preserves lipid ultrastructures at an electron microscopy level. Adoption of polyethylenimine was required for a wide pH range adjustment of the tissue clearing solution. The combination of polyethylenimine and urea had a good tissue clearing ability for multiple tissues within several hours. Blood vessels stained with lipophilic carbocyanine dyes were deeply visible using the solution. Adjusting the pH of the solution was important to maximize the fluorescent intensity and suppress dye leakage during tissue clearing. The spatial distribution of doxorubicin and oxidative stress were observable using the solution. Moreover, spatial distribution of liposomes in the liver was visualized. Hence, the Seebest solution provides pH-adjustable, rapid, sufficient tissue clearing, while preserving lipid ultrastructures, which is suitable for drug delivery system evaluations.
可视化生物事件和状态以解决生物学问题具有挑战性。组织透明化允许进行三维多色成像。在此，我们描述了一种pH可调的组织透明化溶液Seebest（在组织中观察生物事件和状态），它能在电子显微镜水平上保存脂质超微结构。为了对组织透明化溶液进行宽pH范围的调节，需要采用聚乙烯亚胺。聚乙烯亚胺和尿素的组合在数小时内对多种组织具有良好的组织透明化能力。使用该溶液可清晰地看到用亲脂性碳菁染料染色的血管。调节溶液的pH对于在组织透明化过程中最大化荧光强度和抑制染料泄漏很重要。使用该溶液可观察到阿霉素和氧化应激的空间分布。此外，还可视化了脂质体在肝脏中的空间分布。因此，Seebest溶液提供了pH可调、快速、充分的组织透明化，同时保存脂质超微结构，适用于药物递送系统评估。
