Liquid co2

Scanning electron microscopy was performed at the North Carolina State University Center for Electron Microscopy, Raleigh, NC, USA. Agar blocks (~0.5 cm³) containing hyphae were excised and fixed in 0.1 M sodium cacodylate buffer (pH = 6.8) containing 3% glutaraldehyde at 4°C for several weeks. The agar blocks were rinsed with cold 0.1 M sodium cacodylate buffer pH 6.8 three times and then dehydrated in a graded series of ethanol to reach 100% ethanol. The blocks were subjected to critical-point drying with liquid CO₂ (Tousimis Research Corp.) and sputter coated with 50 Å of gold/palladium using a Hummer 6.2 sputter coater (Anatech USA). The samples were viewed at 15 kV with a JSM 5900LV scanning electron microscope (JEOL) and captured with a Digital Scan Generator (JEOL) image acquisition system.

Inoculation and incubation conditions for microscopy specimens were the same as for the other mating crosses. For the light micrographs, a cross of DSM27421 × CBS6039 on V8 (pH = 5) mating medium was incubated for 39 days. The crosses were viewed with a Zeiss Scope.A1 microscope and photographed with a Zeiss Axiocam 105 color camera. For the scanning electron micrographs (SEM), a cross of DSM27421 × CBS6039 on V8 (pH = 5) mating medium was incubated for 11 days. SEM was performed at the North Carolina State University Center for Electron Microscopy, Raleigh, NC, USA. Agar blocks (approximately 0.5 cm³) containing hyphae on the edges of mating patches were excised and fixed in 0.1 M sodium cacodylate buffer (pH = 6.8) containing 3% glutaraldehyde at 4°C for several weeks. Before viewing, the agar blocks were rinsed with cold 0.1 M sodium cacodylate buffer (pH = 6.8) three times and then dehydrated in a graded series of ethanol to reach 100% ethanol. The blocks were subjected to critical-point drying with liquid CO₂ (Tousimis Research Corp.) and sputter coated with 50 Å of gold/palladium using a Hummer 6.2 sputter coater (Anatech USA). The samples were viewed at 15 kV with a JSM 5900LV scanning electron microscope (JEOL) and captured with a Digital Scan Generator (JEOL) image acquisition system.