Bx50f4 light microscope

Immunohistochemistry was performed as previously described [18 (link)]. In short, after dewaxing and rehydration, the slides were immersed in microwave-pretreated boiling citrate buffer (0.01 M, pH 8.0) for antigen repair. After cooling at room temperature (21°C), 3% H₂O₂ was added to deactivate endogenous peroxidase. The slides were washed in phosphate buffer solution and incubated with blocking serum (10% nonimmune goat serum) to block nonspecific immunolabeling at room temperature (21°C). Primary antibodies were incubated with the slides overnight at 4°C. Slides were then incubated with the appropriate secondary antibodies. After rinsing with phosphate buffer solution, the sections were visualized with diaminobenzidine and counterstained with hematoxylin. Neutral gum was used for sealing, and all the sections were photographed under an Olympus BX50F4 light microscope (Olympus, Center Valley, PA).

Sections of stromata were cut by hand using a safety razor blade or with a freezing microtome (ca. 15–30 µm thick) and mounted in either water, 5% KOH, 85% lactic acid, or Lugol’s solution with or without 5% KOH pretreatment to test amyloid reactions⁷⁴ . Stromata and colony colours were described using alphanumeric codes⁷⁵ . Observations of the asexual morph were made from living cultures grown on oatmeal agar (OA)^{76 (link)}. Microscopic measurements were taken from living material mounted in deionized water and are presented as ranges calculated from the mean ± standard deviation of each measured value with outliers in brackets. Observations were made using an Olympus BX50F4 light microscope and an Olympus SZX12 stereo microscope (Olympus, Tokyo, Japan). Images were captured with an InfinityX-32 camera (Lumenera Corp., Ottawa, Canada) using Infinity Analyze v. 6.5.2 (Lumenera Corp.) software. Photographic plates were assembled using Adobe Photoshop CC 2017.1.1 (Adobe Systems, San Jose, California, USA). Cardinal temperatures were assessed for the type strain (DAOMC 252031) by incubating single-point inoculated Petri dishes containing MEA at 5 °C intervals from 5–40 °C. Each treatment was conducted in triplicate and colony diameters were measured two weeks after inoculation.