Live cell fluorescence microscopy to study microbial pathogenesis

  title={Live cell fluorescence microscopy to study microbial pathogenesis},
  author={Adam D. Hoppe and Stephanie Seveau and Joel A. Swanson},
  journal={Cellular Microbiology},
Advances in microscopy and fluorescent probes provide new insight into the nanometer‐scale biochemistry governing the interactions between eukaryotic cells and pathogens. When combined with mathematical modelling, these new technologies hold the promise of qualitative, quantitative and predictive descriptions of these pathways. Using the light microscope to study the spatial and temporal relationships between pathogens, host cells and their respective biochemical machinery requires an… 

N-Way FRET Microscopy of Multiple Protein-Protein Interactions in Live Cells

The N-Way FRET method provides a new toolbox for capturing multiple molecular processes with high spatial and temporal resolution in living cells and is demonstrated by monitoring the oligomerization of three FP-tagged HIV Gag proteins whose tight association in the viral capsid is readily observed.

Contribution of high‐content imaging technologies to the development of anti‐infective drugs

  • M. AngKevin Pethe
  • Biology
    Cytometry. Part A : the journal of the International Society for Analytical Cytology
  • 2016
Recent advances in HCI technologies and chemical biology approaches are reviewed that are contributing to an increased understanding of the intricate host–pathogen interrelationship on the cellular level, and which will foster the development of novel therapeutic approaches for the treatment of human bacterial and protozoan infections.

Deep-red fluorescent imaging probe for bacteria.

Image‐based systems biology of infection

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Noninvasive biophotonic imaging for studies of infectious disease

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Bioimaging structural signatures of the oomycete pathogen Sclerospora graminicola in pearl millet using different microscopic techniques

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Mathematical and computational approaches can complement experimental studies of host–pathogen interactions

Four examples where modelling has complemented current experimental techniques in a way that can or has already pushed knowledge of host–pathogen dynamics forward are outlined.

Emerging trends in aggregation induced emissive luminogens as bacterial theranostics

The initial section of the article reveals the necessity for incorporating diagnostic imaging with antibacterial therapy, while the latter part delivers mechanistic insights on the benefits of AIE fluorophores in theranostic applications.



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The current advancements and knowledge in multiphoton microscopy and how it can be related to the field of microbial infections are detailed.

Three-chromophore FRET microscopy to analyze multiprotein interactions in living cells

Both theoretical and experimental approaches that discriminate the parallel versus the sequential energy transfer processes in the 3-FRET system are described, which highlight the potential of 3- FRET microscopy in studies of spatial and temporal regulation of signaling processes in living cells.

Live-cell photoactivated localization microscopy of nanoscale adhesion dynamics

By allowing observation of a wide variety of nanoscale dynamics, live-cell PALM provides insights into molecular assembly during the initiation, maturation and dissolution of cellular processes.

A guide to accurate fluorescence microscopy colocalization measurements.

Through simulations, imaging of fluorescent antibodies adsorbed on glass and cell measurements, it is shown that ICCS performs much better than standard colocalization algorithms at moderate to high densities of particles, which are often encountered in cellular systems.

Three-dimensional FRET reconstruction microscopy for analysis of dynamic molecular interactions in live cells.

These results verify previous observations that Cdc42 signaling is localized to the advancing margins of forming phagosomes in macrophages and devise and validate a new approach (called 3D-FRET stoichiometry reconstruction, 3DFSR), which provides orders of magnitude gains in both sensitivity and accuracy.

Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy

Large, living biological specimens present challenges to existing optical imaging techniques because of their absorptive and scattering properties. We developed selective plane illumination

Technoreview: Molecular imaging of host–pathogen interactions in intact small animals

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A quantitative evolutionary strategy using fluorescence-activated cell sorting to optimize a cyan-yellow fluorescent protein pair for FRET provides substantially improved sensitivity and dynamic range for a broad range of molecular imaging and screening applications.

Tracking single Kinesin molecules in the cytoplasm of mammalian cells.

By combining direct genetic labeling and single molecule imaging in vivo, this work establishes an important new biophysical method for observing single molecules expressed and localized in the mammalian cytoplasm.