Semi-quantitative analysis methods of tubulin based on the fluorescent images
- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 24
- DOI:
- 10.3321/j.issn:1000-2790.2009.24.001
- Key Word:
- 荧光图像;定量;微管;细胞骨架;分形维数;fluorescent image;semi-qualification;microtubule;cytoskeleton;fraetal dimensions
Abstract： 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.
- [1]姚宇华,严洪,熊江辉.基于分形特征的模拟失重条件下细胞骨架的图像分析[J].2004,(2).
- [2]F. Caserta,W.D. Eldred,E. Fernandez,etc.Determination of fractal dimension of physiologically characterized neurons in two and three dimensions[J].1995,56(2).
- [3]Carmeliet, G.,Nys, G.,Bouillon, R..Microgravity Reduces the Differentiation of Human Osteoblastic MG-63 Cells[J].1997,12(5).
- [4]P R, Montague,M J, Friedlander.Morphogenesis and territorial coverage by isolated mammalian retinal ganglion cells.[J].1991,11(5).
- [5]Karin Paiha,Ilja Vietor,Stefan Thurner,etc.Quantitative measurement of cell migration using time-lapse videomicroscopy and non-linear system analysis[J].2003,119(1).
- [6]Hotchkiss RS,Karl IE,Buchman TG,etc.Stress-induced fractal rearrangement of the endothelial cell cytoskeleton causes apoptosis.[J].1998,124(2).
- [7]D, Ingber.How cells (might) sense microgravity.[J].1999,13 Suppl.
- [8]Rodenacker K,Jutting U,Hughes Fulford M.Reduction of anabolic signals and alteration of osteoblast nuclear morphology in microgravity.[J].2006,99(2).
- [9]Yuval Gazit,James W. Baish,Nina Safabakhsh,etc.Fractal Characteristics of Tumor Vascular Architecture During Tumor Growth and Regression[J].1997,4(4).
- [10]M, Braun,A, Sievers.Role of the microtubule cytoskeleton in gravisensing Chara rhizoids.[J].1994,63(2).
- [11]D B, Thomason,O, Anderson,V, Menon.Fractal analysis of cytoskeleton rearrangement in cardiac muscle during head-down tilt.[J].1996,81(4).
- [12]Shang,P.,Hu,L.-F.,Tian,Z.-C.,etc.Large gradient high magnetic field affects the association of MACF1 with actin and microtubule cytoskeleton[J].2009,30(7).
- [13]Sedivy R,Zielinski CC,Kostler WJ,etc.Short-term rhythmic proliferation of human breast cancer cell lines: surface effects and fractal growth patterns.[J].2002,197(2).
- [14]Moran EL,Caldwell CB,Bogoch ER.Fractal dimension as a measure of altered trabecular bone in experimental inflammatory arthritis.[J].1998,13(6).