IMMUNOFLUORESCENCE PHASE SENSITIVE FLOW CYTOMETRY: CELL CYCLE

该项目的具体目标是增加 免疫荧光流式细胞术信噪比（S/N） 细胞周期调节分子的检测 用相敏检波消除背景噪声源 方法. 目前，在大细胞中不能检测到p53，例如， 成纤维细胞和上皮细胞，由于背景噪声 与激光散射、拉曼散射、非特异性探针相关 结合和内在细胞背景自发荧光。 我们将 研究使用相敏流式细胞术来减少或 消除这些类型的干扰，从而提高信噪比 低水平的测量。 目前正在进行实验， 确定未染色（固定）成纤维细胞和细胞的寿命 碘化丙啶和增殖核抗原标记 （PCNA-FITC），并进行DNA的相位分辨发射测量 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.