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Analysis of Grayscale Characteristics in Images of Labeled Microtubules from Cultured Cardiac Myocytes

Published online by Cambridge University Press:  16 March 2015

Yongming Dang ,
Xiaodong Lan ,
Qiong Zhang ,
Lingfei Li  and
Yuesheng Huang
Yongming Dang
Affiliation:
State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burn Research, Southwest Hospital, The Third Military Medical University, Chongqing 400038, China
Xiaodong Lan
Affiliation:
State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burn Research, Southwest Hospital, The Third Military Medical University, Chongqing 400038, China
Qiong Zhang
Affiliation:
State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burn Research, Southwest Hospital, The Third Military Medical University, Chongqing 400038, China
Lingfei Li
Affiliation:
State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burn Research, Southwest Hospital, The Third Military Medical University, Chongqing 400038, China
Yuesheng Huang*
Affiliation:
State Key Laboratory of Trauma, Burns and Combined Injury, Institute of Burn Research, Southwest Hospital, The Third Military Medical University, Chongqing 400038, China
*
*Corresponding author. yshuang.tmmu@gmail.com
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Abstract

Microtubules of cardiac myocytes depolymerize after a hypoxic insult or treatment with colchicine. However, little attention has been paid to quantifying changes in microtubule distribution when using fluorescent images. We converted fluorescence images of labeled microtubules in H9C2 cardiac myocytes to grayscale images, then filtered the images to remove any noise, and used grayscale histograms to quantify features of the images. The results show that parameters such as the mean, variance, skewness, kurtosis, energy, and entropy can be used to quantitatively describe the distribution of microtubules in cells. Quantitative characteristics of microtubule distribution were similar after culturing cells under hypoxic conditions or after treatment with colchicine. These results parallel those described for neonatal rat cardiac myocytes following ischemia and hypoxia. In addition, we provide a method for internal segmentation of the cells, which revealed that microtubular depolymerization was more evident near the cell membrane following hypoxia or colchicine treatment.

Keywords

fluorescencehypoxiacolchicinequantificationdepolymerization
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 21 , Issue 2 , April 2015 , pp. 334 - 342
Copyright
© Microscopy Society of America 2015 

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