FLUORESCENT THIOL PROBES - CYTOMETRY AND PCR TECHNOLOGY

荧光硫醇探针 - 细胞计数和 PCR 技术

• 批准号: 3493108
• 负责人: MARGARET E LANGMUIR
• 金额: $ 5万
• 依托单位: COVALENT ASSOCIATES, INC.
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-06-02 至 1992-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3493108
• 关键词: chemical kinetics; chemical synthesis; cytotoxicity; emission spectrometry; flow cytometry; fluorescent dye /probe; glutathione; ionophores; maleimides; membrane permeability; polymerase chain reaction; thiols; tissue /cell culture

We propose to synthesize new probes for the determination of intracellular glutathione and other cellular thiols. The reaction product of the probe with thiols will have longer wavelength absorption and emission and higher quantum yield than the best cytosolic thiol fluorophore currently in use for cell sorting by flow cytometry. The dye will have the advantage of being cell membrane permeable so that cell loading can occur at lower extra cellular dye concentrations than currently possible. The synthesized probes will be tested in vitro and in cell cultures for absorption and emission properties when bound to glutathione and other protein thiols. Cell toxicity will also be determined.

我们建议合成新的探针用于测定细胞内的 谷胱甘肽和其他细胞硫醇。 探针的反应产物 硫醇将具有更长的波长吸收和发射， 量子产率比目前使用的最好的胞质硫醇荧光团 用于通过流式细胞术进行细胞分选。 这种染料的优点是 是细胞膜可渗透的，使得细胞负载可以在较低的额外温度下发生， 细胞染料浓度比目前可能的。 合成的 将在体外和细胞培养物中测试探针的吸收， 当与谷胱甘肽和其他蛋白质硫醇结合时，发射特性。 还将测定细胞毒性。