Difference of EGCg adhesion on cell surface between Staphylococcus aureus and Escherichia coli visualized by electron microscopy after novel indirect staining with cerium chloride

Motokazu Nakayama, Naofumi Shigemune, Takashi Tsugukuni, Hajime Tokuda, Takahisa Miyamoto

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

We developed a novel method using indirect staining with cerium chloride for visualization of the catechin derivative epigallocatechin gallate (EGCg) on the surface of particles, i.e., polystyrene beads and bacterial cells, by electron microscopy. The staining method is based on the fact that in an alkaline environment, EGCg produces hydrogen peroxide, and then hydrogen peroxide reacts with cerium, resulting in a cerium hydroperoxide precipitate. This precipitate subsequently reacts with EGCg to produce larger deposits. The amount of precipitate is proportional to the amount of EGCg. Highly EGCg-sensitive Staphylococcus aureus and EGCg-resistant Escherichia coli were treated with EGCg under various pH conditions. Transmission electron microscopy observation showed that the amount of deposits on S. aureus increased with an increase in EGCg concentration. After treating bacterial cells with 0.5. mg/mL EGCg (pH 6.0), attachment of EGCg was significantly lower to E. coli than to S. aureus. This is the first report that shows differences in affinity of EGCg to the cell surfaces of Gram-positive and -negative bacteria by electron microscopy.

Original languageEnglish
Pages (from-to)97-103
Number of pages7
JournalJournal of Microbiological Methods
Volume86
Issue number1
DOIs
Publication statusPublished - Jul 1 2011

Fingerprint

Cerium
Cell Adhesion
Staphylococcus aureus
Chlorides
Electron Microscopy
Staining and Labeling
Escherichia coli
Hydrogen Peroxide
epigallocatechin gallate
Catechin
Polystyrenes
Gram-Positive Bacteria
Gram-Negative Bacteria
Transmission Electron Microscopy

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

Cite this

Difference of EGCg adhesion on cell surface between Staphylococcus aureus and Escherichia coli visualized by electron microscopy after novel indirect staining with cerium chloride. / Nakayama, Motokazu; Shigemune, Naofumi; Tsugukuni, Takashi; Tokuda, Hajime; Miyamoto, Takahisa.

In: Journal of Microbiological Methods, Vol. 86, No. 1, 01.07.2011, p. 97-103.

Research output: Contribution to journalArticle

@article{65a2d91ae44a4957a4defea60bc72a7b,
title = "Difference of EGCg adhesion on cell surface between Staphylococcus aureus and Escherichia coli visualized by electron microscopy after novel indirect staining with cerium chloride",
abstract = "We developed a novel method using indirect staining with cerium chloride for visualization of the catechin derivative epigallocatechin gallate (EGCg) on the surface of particles, i.e., polystyrene beads and bacterial cells, by electron microscopy. The staining method is based on the fact that in an alkaline environment, EGCg produces hydrogen peroxide, and then hydrogen peroxide reacts with cerium, resulting in a cerium hydroperoxide precipitate. This precipitate subsequently reacts with EGCg to produce larger deposits. The amount of precipitate is proportional to the amount of EGCg. Highly EGCg-sensitive Staphylococcus aureus and EGCg-resistant Escherichia coli were treated with EGCg under various pH conditions. Transmission electron microscopy observation showed that the amount of deposits on S. aureus increased with an increase in EGCg concentration. After treating bacterial cells with 0.5. mg/mL EGCg (pH 6.0), attachment of EGCg was significantly lower to E. coli than to S. aureus. This is the first report that shows differences in affinity of EGCg to the cell surfaces of Gram-positive and -negative bacteria by electron microscopy.",
author = "Motokazu Nakayama and Naofumi Shigemune and Takashi Tsugukuni and Hajime Tokuda and Takahisa Miyamoto",
year = "2011",
month = "7",
day = "1",
doi = "10.1016/j.mimet.2011.04.010",
language = "English",
volume = "86",
pages = "97--103",
journal = "Journal of Microbiological Methods",
issn = "0167-7012",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Difference of EGCg adhesion on cell surface between Staphylococcus aureus and Escherichia coli visualized by electron microscopy after novel indirect staining with cerium chloride

AU - Nakayama, Motokazu

AU - Shigemune, Naofumi

AU - Tsugukuni, Takashi

AU - Tokuda, Hajime

AU - Miyamoto, Takahisa

PY - 2011/7/1

Y1 - 2011/7/1

N2 - We developed a novel method using indirect staining with cerium chloride for visualization of the catechin derivative epigallocatechin gallate (EGCg) on the surface of particles, i.e., polystyrene beads and bacterial cells, by electron microscopy. The staining method is based on the fact that in an alkaline environment, EGCg produces hydrogen peroxide, and then hydrogen peroxide reacts with cerium, resulting in a cerium hydroperoxide precipitate. This precipitate subsequently reacts with EGCg to produce larger deposits. The amount of precipitate is proportional to the amount of EGCg. Highly EGCg-sensitive Staphylococcus aureus and EGCg-resistant Escherichia coli were treated with EGCg under various pH conditions. Transmission electron microscopy observation showed that the amount of deposits on S. aureus increased with an increase in EGCg concentration. After treating bacterial cells with 0.5. mg/mL EGCg (pH 6.0), attachment of EGCg was significantly lower to E. coli than to S. aureus. This is the first report that shows differences in affinity of EGCg to the cell surfaces of Gram-positive and -negative bacteria by electron microscopy.

AB - We developed a novel method using indirect staining with cerium chloride for visualization of the catechin derivative epigallocatechin gallate (EGCg) on the surface of particles, i.e., polystyrene beads and bacterial cells, by electron microscopy. The staining method is based on the fact that in an alkaline environment, EGCg produces hydrogen peroxide, and then hydrogen peroxide reacts with cerium, resulting in a cerium hydroperoxide precipitate. This precipitate subsequently reacts with EGCg to produce larger deposits. The amount of precipitate is proportional to the amount of EGCg. Highly EGCg-sensitive Staphylococcus aureus and EGCg-resistant Escherichia coli were treated with EGCg under various pH conditions. Transmission electron microscopy observation showed that the amount of deposits on S. aureus increased with an increase in EGCg concentration. After treating bacterial cells with 0.5. mg/mL EGCg (pH 6.0), attachment of EGCg was significantly lower to E. coli than to S. aureus. This is the first report that shows differences in affinity of EGCg to the cell surfaces of Gram-positive and -negative bacteria by electron microscopy.

UR - http://www.scopus.com/inward/record.url?scp=79956275090&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79956275090&partnerID=8YFLogxK

U2 - 10.1016/j.mimet.2011.04.010

DO - 10.1016/j.mimet.2011.04.010

M3 - Article

VL - 86

SP - 97

EP - 103

JO - Journal of Microbiological Methods

JF - Journal of Microbiological Methods

SN - 0167-7012

IS - 1

ER -