---
_id: '503'
abstract:
- lang: eng
  text: Buffers are essential for diluting bacterial cultures for flow cytometry analysis
    in order to study bacterial physiology and gene expression parameters based on
    fluorescence signals. Using a variety of constitutively expressed fluorescent
    proteins in Escherichia coli K-12 strain MG1655, we found strong artifactual changes
    in fluorescence levels after dilution into the commonly used flow cytometry buffer
    phosphate-buffered saline (PBS) and two other buffer solutions, Tris-HCl and M9
    salts. These changes appeared very rapidly after dilution, and were linked to
    increased membrane permeability and loss in cell viability. We observed buffer-related
    effects in several different E. coli strains, K-12, C and W, but not E. coli B,
    which can be partially explained by differences in lipopolysaccharide (LPS) and
    outer membrane composition. Supplementing the buffers with divalent cations responsible
    for outer membrane stability, Mg2+ and Ca2+, preserved fluorescence signals, membrane
    integrity and viability of E. coli. Thus, stabilizing the bacterial outer membrane
    is essential for precise and unbiased measurements of fluorescence parameters
    using flow cytometry.
acknowledged_ssus:
- _id: Bio
acknowledgement: "We thank R Chait and M Lagator for sharing Bacillus subtilis CR_Y1
  and pZS*_2R-cIPtet-Venus-Prm, respectively. We are grateful to T Pilizota and all
  members of the Guet lab for critically reading the manuscript. We also thank the
  Bioimaging facility at IST Austria for assistance using the FACSAria III system.\r\n\r\n"
article_processing_charge: No
author:
- first_name: Kathrin
  full_name: Tomasek, Kathrin
  id: 3AEC8556-F248-11E8-B48F-1D18A9856A87
  last_name: Tomasek
  orcid: 0000-0003-3768-877X
- first_name: Tobias
  full_name: Bergmiller, Tobias
  id: 2C471CFA-F248-11E8-B48F-1D18A9856A87
  last_name: Bergmiller
  orcid: 0000-0001-5396-4346
- first_name: Calin C
  full_name: Guet, Calin C
  id: 47F8433E-F248-11E8-B48F-1D18A9856A87
  last_name: Guet
  orcid: 0000-0001-6220-2052
citation:
  ama: Tomasek K, Bergmiller T, Guet CC. Lack of cations in flow cytometry buffers
    affect fluorescence signals by reducing membrane stability and viability of Escherichia
    coli strains. <i>Journal of Biotechnology</i>. 2018;268:40-52. doi:<a href="https://doi.org/10.1016/j.jbiotec.2018.01.008">10.1016/j.jbiotec.2018.01.008</a>
  apa: Tomasek, K., Bergmiller, T., &#38; Guet, C. C. (2018). Lack of cations in flow
    cytometry buffers affect fluorescence signals by reducing membrane stability and
    viability of Escherichia coli strains. <i>Journal of Biotechnology</i>. Elsevier.
    <a href="https://doi.org/10.1016/j.jbiotec.2018.01.008">https://doi.org/10.1016/j.jbiotec.2018.01.008</a>
  chicago: Tomasek, Kathrin, Tobias Bergmiller, and Calin C Guet. “Lack of Cations
    in Flow Cytometry Buffers Affect Fluorescence Signals by Reducing Membrane Stability
    and Viability of Escherichia Coli Strains.” <i>Journal of Biotechnology</i>. Elsevier,
    2018. <a href="https://doi.org/10.1016/j.jbiotec.2018.01.008">https://doi.org/10.1016/j.jbiotec.2018.01.008</a>.
  ieee: K. Tomasek, T. Bergmiller, and C. C. Guet, “Lack of cations in flow cytometry
    buffers affect fluorescence signals by reducing membrane stability and viability
    of Escherichia coli strains,” <i>Journal of Biotechnology</i>, vol. 268. Elsevier,
    pp. 40–52, 2018.
  ista: Tomasek K, Bergmiller T, Guet CC. 2018. Lack of cations in flow cytometry
    buffers affect fluorescence signals by reducing membrane stability and viability
    of Escherichia coli strains. Journal of Biotechnology. 268, 40–52.
  mla: Tomasek, Kathrin, et al. “Lack of Cations in Flow Cytometry Buffers Affect
    Fluorescence Signals by Reducing Membrane Stability and Viability of Escherichia
    Coli Strains.” <i>Journal of Biotechnology</i>, vol. 268, Elsevier, 2018, pp.
    40–52, doi:<a href="https://doi.org/10.1016/j.jbiotec.2018.01.008">10.1016/j.jbiotec.2018.01.008</a>.
  short: K. Tomasek, T. Bergmiller, C.C. Guet, Journal of Biotechnology 268 (2018)
    40–52.
corr_author: '1'
date_created: 2018-12-11T11:46:50Z
date_published: 2018-02-20T00:00:00Z
date_updated: 2024-10-09T20:58:29Z
day: '20'
department:
- _id: CaGu
doi: 10.1016/j.jbiotec.2018.01.008
external_id:
  isi:
  - '000425715100006'
intvolume: '       268'
isi: 1
language:
- iso: eng
month: '02'
oa_version: None
page: 40 - 52
publication: Journal of Biotechnology
publication_status: published
publisher: Elsevier
publist_id: '7317'
quality_controlled: '1'
scopus_import: '1'
status: public
title: Lack of cations in flow cytometry buffers affect fluorescence signals by reducing
  membrane stability and viability of Escherichia coli strains
type: journal_article
user_id: c635000d-4b10-11ee-a964-aac5a93f6ac1
volume: 268
year: '2018'
...