Journal: PLoS Pathogens

Article Title: Distinct Roles for Dectin-1 and TLR4 in the Pathogenesis of Aspergillus fumigatus Keratitis

doi: 10.1371/journal.ppat.1000976

Figure Lengend Snippet: A mouse model of Aspergillus keratitis was developed by injecting 1×10 5 Af293.1RFP conidia into the corneal stroma, and tracking disease progression over time. A. Slit-lamp microscopy of immunocompetent and cyclophosphamide-immunosuppressed (Cyc) C57BL/6J corneas infected for 24, 48, and 72h. Fluorescence microscopy of emitted 580nm light reveals fungal growth in the corresponding cornea. B. 5µm paraffin section of the central cornea were stained with Periodic Acid Schiff and Hematoxylin (PASH) to visualize cellular infiltration, fungal growth, and corneal pathology at 24, 48, and 72 h post-infection. C. 5µm paraffin sections were immunostained with anti-neutrophil NIMPR-14 to visualize neutrophil infiltration at 24, 48, and 72 h post-infection. D. Bright-light reflected from the cornea, and 580nm light emitted from the cornea were independently captured as TIFF images and the pixel intensity of each image was analyzed within a constant defined region encompassing the cornea to quantify corneal opacity or fungal RFP expression (corresponding to fungal biomass), respectively. E. Corneal opacity in immunocompetent and cyc-immunosuppressed C57BL/6 mice at 24–72h post-infection F. Fungal RFP expression in immunocompetent and cyc-immunosuppressed C57BL/6 mice at 24 and 72h post-infection. G. At each time point, eyes were enucleated, homogenized, and fungal viability assessed via serial dilutions on Sabouraud Dextrose agar. Data are representative of two independent experiments, with five mice per time point.

Article Snippet: Subsequently, serial log dilutions were performed and plated onto bacteriologic-grade Sabouraud dextrose agar plates (Becton Dickenson).

Techniques: Biomarker Discovery, Microscopy, Infection, Fluorescence, Paraffin Section, Staining, Expressing