title: Hyperspectral Imaging Using Intracellular Spies: Quantitative Real-Time Measurement of Intracellular Parameters In Vivo during Interaction of the Pathogenic Fungus Aspergillus fumigatus with Human Monocytes.
abstract: Hyperspectral imaging (HSI) is a technique based on the combination of classical spectroscopy and conventional digital image processing. It is also well suited for the biological assays and quantitative real-time analysis since it provides spectral and spatial data of samples. The method grants detailed information about a sample by recording the entire spectrum in each pixel of the whole image. We applied HSI to quantify the constituent pH variation in a single infected apoptotic monocyte as a model system. Previously, we showed that the human-pathogenic fungus Aspergillus fumigatus conidia interfere with the acidification of phagolysosomes. Here, we extended this finding to monocytes and gained a more detailed analysis of this process. Our data indicate that melanised A. fumigatus conidia have the ability to interfere with apoptosis in human monocytes as they enable the apoptotic cell to recover from mitochondrial acidification and to continue with the cell cycle. We also showed that this ability of A. fumigatus is dependent on the presence of melanin, since a non-pigmented mutant did not stop the progression of apoptosis and consequently, the cell did not recover from the acidic pH. By conducting the current research based on the HSI, we could measure the intracellular pH in an apoptotic infected human monocyte and show the pattern of pH variation during 35 h of measurements. As a conclusion, we showed the importance of melanin for determining the fate of intracellular pH in a single apoptotic cell.
authors.affiliation: Department of Cell and Molecular Biology, Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute, Jena, Germany.|Friedrich Schiller University, Jena, Germany.
authors.forename: Sara
authors.initials: S
authors.lastname: Mohebbi
authors.affiliation: Department of Photonics and Sensorics of Jena, Felsbachstrasse, Jena, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Erfurth
authors.affiliation: Department of Cell and Molecular Biology, Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute, Jena, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Hennersdorf
authors.affiliation: Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute, Jena, Germany.|Friedrich Schiller University, Jena, Germany.
authors.forename: Axel A
authors.initials: AA
authors.lastname: Brakhage
authors.affiliation: Department of Cell and Molecular Biology, Leibniz Institute for Natural Product Research and Infection Biology - Hans Knöll Institute, Jena, Germany.|Friedrich Schiller University, Jena, Germany.
authors.forename: Hans Peter
authors.initials: HP
authors.lastname: Saluz
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001230:Aspergillus / Q000378:metabolism* / Q000472:pathogenicity; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000737:chemistry; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D054884:Host-Pathogen Interactions; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018942:Macrolides / Q000494:pharmacology; D008543:Melanins / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D009000:Monocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000382:microbiology*; D016296:Mutagenesis; D010587:Phagocytosis / Q000187:drug effects; D010588:Phagosomes / Q000187:drug effects / Q000378:metabolism; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013172:Spores, Fungal / Q000737:chemistry / Q000187:drug effects / Q000502:physiology
chemical_list: D005456:Fluorescent Dyes; D005656:Fungal Proteins; D018942:Macrolides; D008543:Melanins; D015638:Cytochalasin D; D002738:Chloroquine; C052502:bafilomycin C1