A rapid and convenient method for fluorescence analysis of in vitro cultivated metacestode vesicles from Echinococcus multilocularis

Zhe Cheng; Fan Liu; Shan Zhu; Huimin Tian; Liang Wang; Yanhai Wang

doi:10.1371/journal.pone.0118215

A rapid and convenient method for fluorescence analysis of in vitro cultivated metacestode vesicles from Echinococcus multilocularis

PLoS One. 2015 Feb 23;10(2):e0118215. doi: 10.1371/journal.pone.0118215. eCollection 2015.

Authors

Zhe Cheng¹, Fan Liu², Shan Zhu¹, Huimin Tian², Liang Wang¹, Yanhai Wang¹

Affiliations

¹ State Key Laboratory of Cellular Stress Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian, China.
² Medical College, Xiamen University, Xiamen, Fujian, China.

Abstract

We here describe a convenient method for preparation, fixation and fluorescence analysis of in vitro cultivated metacestode vesicles from E. multilocularis. Parasite materials could be prepared in one hour, did not need to be sectioned, and were subsequently utilized for further whole-mount staining assays directly. Using these preparations, in combination with conventional fluorescence staining techniques, we could detect the expression and subcellular localization of a specific protein and identify in situ proliferative or apoptotic cells in the germinal layer of metacestode vesicles. Based on this approach, future molecular and cellular analysis of Echinococcus metacestode vesicles in the in vitro system will be greatly facilitated.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis
Cell Proliferation
Echinococcus multilocularis / cytology
Echinococcus multilocularis / metabolism*
Extracellular Signal-Regulated MAP Kinases / metabolism
Fluorescent Antibody Technique / methods*
Helminth Proteins / metabolism*
Humans
Larva / cytology
Larva / metabolism
Microscopy, Fluorescence / methods*
Phosphorylation
Reproducibility of Results
Time Factors
Tubulin / metabolism

Substances

Helminth Proteins
Tubulin
Extracellular Signal-Regulated MAP Kinases

Grants and funding

This work was supported by the Fundamental Research Funds for the Central Universities of People's Republic of China (No. 2012121044) and the National Natural Science Foundation of China (No. 81000740, 81171596, 81201305, 81471970) (http://www.nsfc.gov.cn/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.