Semi-quantitative analysis methods of tubulin based on the fluorescent images
AIM:To establish a quick,convenient and semiquantitative method based on fluorescent images for microtubule cytoskeleton analysis.METHODS:Three image analysis software including SimplePCI,Image J and Image-pro plus 6.0 (IPP),and Western Blot assays were used to qualify the microtubale cytoskeleton alterations in MC3T3-E1 cell after 48 h of being cultured under Radom Position Machine(3D)condition.The Fractal dimensions(D)of cytoskeleton architecture of MC3T3-E1 were calculated by box counting method using Image J.RESULTS:After being cultured for 48 h in RPM,the Fluorescent intensity(grey level)of MC3T3-E1 cytoskeleton was lower than that under control condition(P<0.01).Western Blot analysis showed after being cultured for 48 h in RPM,α-tubulin in MC3T3-E1 cells expression was decreased significantly(P<0.001),and the fractal dimension was also lower than that under control condition(P<0.05).CONCLUSION:The method based on fluorescent Images used in this study is useful to qualify both the location and expression of mierotubule cytoskeleton,which provides a new insight into the quantitative morphological study of cytoskeleton.
NETL
NSTL
万方数据
维普
