LASER BASED CELL LOADING & LYSIS: SINGLE CELL ELECTROPHORESIS: KINASE ACTIVITY

基于激光的细胞装载

• 批准号: 6220404
• 负责人: Nancy L. Allbritton
• 金额: $ 3.09万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6220404
• 关键词: Mammalia; biological products; biomedical resource; drug /agent; human tissue; injury; lasers; model design /development; skin; technology /technique

Significant progress has been made toward specific aim 3. Currently, single adherent cells are lysed in less than 33 msec by a laser-induced shock wave. The contents of single cells are immediately loaded into a capillary, separated by electrophoresis within the capillary, and detected by laser-induced fluorescence. This was demonstrated and published in 1998 in Analytical Chemistry, vol 70, number 21, pages 4570-4577. Preliminary results show that kinase-substrate peptides can be measured from single mammalian cells. Further, cellular kinase activity can be measured as changes in the degree of substrate phosphorylation. Experiments are in progress to define parameters for and demonstrate physiologically relevant measurements. Specific aims 1 and 2 are yet to be addressed substantively, but will be accelerated by the completion of the current studies on adherent cells. Specific aim 4 will become possible immediately thereafter.

具体目标3已取得重大进展。 目前，单个贴壁细胞可以在不到 33 毫秒的时间内被裂解 激光诱导冲击波。 单细胞内容物为 立即装入毛细管中，通过电泳分离 在毛细管内，并通过激光诱导荧光进行检测。 1998 年《分析化学》对此进行了论证并发表， 第 70 卷，第 21 期，第 4570-4577 页。 初步结果表明 激酶底物肽可以从单个哺乳动物细胞中测量。 此外，细胞激酶活性可以通过 底物磷酸化程度。 实验正在进行中 定义参数并证明生理相关 测量。 具体目标 1 和 2 尚待解决 实质上，但将随着完成 目前对贴壁细胞的研究。 具体目标 4 将变为 此后立即可能。