IMMUNOFLUORESCENCE PHASE SENSITIVE FLOW CYTOMETRY: CELL CYCLE

这个项目的具体目标是增加 免疫荧光流式细胞术的信噪比(S/N) 细胞内细胞周期调节分子的检测 相敏检测法消除背景噪声源 方法：研究方法。目前，在大细胞中无法检测到P53，例如， 成纤维细胞和上皮细胞，因为背景噪声 与激光散射、拉曼散射、非特异性探针相关 结合，和固有的细胞背景自体荧光。我们会 研究用相敏流式细胞术减少或减少 消除这些类型的干扰，从而增加S/N的比率 在低层测量中。目前正在进行实验，以 测定未染色(固定)的成纤维细胞和细胞的寿命 碘化丙啶标记增殖性核抗原 (增殖细胞核抗原-FITC)和进行DNA的相分辨发射测量 含量和免疫荧光。

The specific aim of this project is to increase the signal-to-noise ratio (S/N) for immunofluorescence flow cytometry detection of intracellular cell cycle regulator molecules by eliminating sources of background noise by phase-sensitive detection methods. At present, p53 cannot be detected in large cells, e.g., fibroblasts and epithelial cell, because of the background noise associated with laser scatter, Raman scatter, nonspecific probe binding, and intrinsic cellular background autofluorescence. We will investigate the use of phase-sensitive flow cytometry to reduce or eliminate these types of interference and thus increase the S/N ratio in low-level measurements. Experiments are currently underway to determine the lifetimes of unstained (fixed) fibroblasts and cells labeled with propidium iodide and proliferating nuclear antigen (PCNA-FITC) and to perform phase-resolved emission measurements of DNA content and immunofluorescence.