Leitner F, Paillasson S, Ronot X, Demongeot J
Laboratoire TIMC, Faculte de Medecine, Universite Joseph Fourier, Institut Albert Bonniot, Domaine de la Merci, La Tronche, France.
Acta Biotheor. 1995 Dec;43(4):299-317. doi: 10.1007/BF00713555.
Increasing interest has been paid to applications of fluorescence measurements to analyze physiological mechanisms in living cells. However, few studies have taken advantage of DNA quantification by fluorometry for dynamic assessment of chromatin organization as well as cell motion during the cell cycle. This approach involves both optimal conditions for DNA staining and cell tracking methods. In this context, this report describes a stoichiometric method for nuclear DNA specific staining, using the bisbenzimidazole dye Hoechst 33342 associated with verapamil, a calcium membrane channel blocker. This method makes it possible to correlate variations of nuclear DNA content with cell motion in cells that are maintained alive. Motion measurement is the second goal of this paper and it explains the snake-spline method, and the associated cell following method.
荧光测量在分析活细胞生理机制中的应用已受到越来越多的关注。然而，利用荧光法进行DNA定量分析以动态评估细胞周期中染色质组织以及细胞运动的研究却很少。这种方法既涉及DNA染色的最佳条件，也涉及细胞追踪方法。在此背景下，本报告描述了一种使用双苯并咪唑染料Hoechst 33342与钙膜通道阻滞剂维拉帕米相结合的化学计量方法，用于细胞核DNA特异性染色。该方法能够将细胞核DNA含量的变化与存活细胞中的细胞运动联系起来。运动测量是本文的第二个目标，文中解释了蛇形样条法以及相关的细胞跟踪方法。
