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@ARTICLE{Krmer:819825,
      author       = {Krämer, Christina and Wiechert, Wolfgang and Kohlheyer,
                      Dietrich},
      title        = {{T}ime-resolved, single-cell analysis of induced and
                      programmed cell death via non-invasive propidium iodide and
                      counterstain perfusion},
      journal      = {Scientific reports},
      volume       = {6},
      issn         = {2045-2322},
      address      = {London},
      publisher    = {Nature Publishing Group},
      reportid     = {FZJ-2016-05412},
      pages        = {32104},
      year         = {2016},
      abstract     = {Conventional propidium iodide (PI) staining requires the
                      execution of multiple steps prior to analysis, potentially
                      affecting assay results as well as cell vitality. In this
                      study, this multistep analysis method has been transformed
                      into a single-step, non-toxic, real-time method via
                      live-cell imaging during perfusion with 0.1 μM PI inside
                      a microfluidic cultivation device. Dynamic PI staining was
                      an effective live/dead analytical tool and demonstrated
                      consistent results for single-cell death initiated by direct
                      or indirect triggers. Application of this method for the
                      first time revealed the apparent antibiotic tolerance of
                      wild-type Corynebacterium glutamicum cells, as indicated by
                      the conversion of violet fluorogenic calcein acetoxymethyl
                      ester (CvAM). Additional implementation of this method
                      provided insight into the induced cell lysis of Escherichia
                      coli cells expressing a lytic toxin-antitoxin module,
                      providing evidence for non-lytic cell death and cell
                      resistance to toxin production. Finally, our dynamic PI
                      staining method distinguished necrotic-like and
                      apoptotic-like cell death phenotypes in Saccharomyces
                      cerevisiae among predisposed descendants of
                      nutrient-deprived ancestor cells using PO-PRO-1 or green
                      fluorogenic calcein acetoxymethyl ester (CgAM) as
                      counterstains. The combination of single-cell cultivation,
                      fluorescent time-lapse imaging, and PI perfusion facilitates
                      spatiotemporally resolved observations that deliver new
                      insights into the dynamics of cellular behaviour.},
      cin          = {IBG-1},
      ddc          = {000},
      cid          = {I:(DE-Juel1)IBG-1-20101118},
      pnm          = {581 - Biotechnology (POF3-581)},
      pid          = {G:(DE-HGF)POF3-581},
      typ          = {PUB:(DE-HGF)16},
      UT           = {WOS:000382366300001},
      pubmed       = {pmid:27580964},
      doi          = {10.1038/srep32104},
      url          = {https://juser.fz-juelich.de/record/819825},
}