Document Detail


Construction of time-lapse scanning electrochemical microscopy with temperature control and its application to evaluate the preservation effects of antifreeze proteins on living cells.
MedLine Citation:
PMID:  19551995     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Antifreeze proteins (AFPs) can protect cells from hypothermic damage; however, their mechanism of action remains unclear. Scanning electrochemical microscopy (SECM) can evaluate the size and activities of cells, although long-term continuous monitoring has been unsuccessful. We constructed a novel, fully automated, time-lapse SECM system and investigated the cell preservation effect of AFPs by analyzing single cellular topography at low temperatures. From the SECM measurements, mammalian cells (HepG2), treated in Euro-Collins (EC) solution at 4 degrees C, began to swell at 8 h and then immediately ruptured. In AFP-containing EC solution, the cellular size did not change until 16 h and then gradually increased and finally ruptured. In addition, the cellular height at rupture point significantly increased in the presence of AFPs. These results suggest that AFPs stabilize the cellular membrane and protect cells from hypothermic damage. This SECM system allowed us to observe the single cellular response to hypothermia by long-term automatic scanning and will be applicable for analysis to other cellular activities and topographies.
Authors:
Yu Hirano; Yoshiyuki Nishimiya; Keiko Kowata; Fumio Mizutani; Sakae Tsuda; Yasuo Komatsu
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  80     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  2008 Dec 
Date Detail:
Created Date:  2009-06-24     Completed Date:  2009-08-17     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  9349-54     Citation Subset:  IM    
Affiliation:
Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), 2-17-2-1 Tsukisamu-Higashi, Toyohira, Sapporo, Japan. yu.hirano@aist.go.jp
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MeSH Terms
Descriptor/Qualifier:
Animals
Antifreeze Proteins / pharmacology*
Cell Line, Tumor
Cell Size
Cells / cytology,  drug effects*
Cold Temperature
Cryoprotective Agents / pharmacology
Electrochemistry / instrumentation*,  methods
Equipment Design
Fish Proteins / pharmacology*
Humans
Microscopy / instrumentation*,  methods
Perciformes*
Chemical
Reg. No./Substance:
0/Antifreeze Proteins; 0/Cryoprotective Agents; 0/Fish Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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