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

在具体目标3方面取得了重大进展。 目前，单个贴壁细胞在不到33毫秒内被裂解， 激光诱导冲击波 单个细胞的内容物是 立即加载到毛细管中，通过电泳分离 在毛细管内，并通过激光诱导荧光检测。 这在1998年的《分析化学》上得到了证实和发表， 第70卷第21期第4570-4577页。 初步结果显示 激酶底物肽可以从单个哺乳动物细胞测量。 此外，细胞激酶活性可以测量为细胞内激酶活性的变化。 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性 测量. 具体目标1和2尚待解决 这一进程将在2010年完成， 贴壁细胞的研究现状。 具体目标4将成为 可能之后立即。

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.