Axiocam erc5

In order to assess the ectomycorrhizal community structure, fine roots were removed from each soil core taken in 2014 to give a sample with approximately 150–300 root tips per core. The samples were then washed carefully, placed into petri-dishes filled with clean tap water, and stored at 4 °C (analyzed within three weeks). All clearly definable ectomycorrhizal root tips from each sample were sorted into morphotypes based on the method described by Agerer (1997 ), using a ZEISS (Stemi 2000-CS) dissecting microscope which was connected with an AxioCam ERc5s camera. The final identification to genus or species level (where possible) was carried out by sequencing of DNA (see below). The total number of root tips colonized by each of the morphotype was counted under the dissecting microscope. Between 1 and 10 ectomycorrhizal root tips of each morphotype were placed into micro-centrifuge tubes. The number of root tips varied from one to ten depending on the abundance of the morphotype. The samples were then stored at −20 °C until DNA extraction.

The Alcian blue-periodic acid Schiff (AB-PAS) staining technique was used, as previously described [54 (link)], to identify mucous-secreting cells in the bronchiolar epithelium. Photomicrographs at x400 were taken using a light microscope equipped with a digital camera (Axiocam ERc5s). A total of 20–30 bronchioles (900–1700 μm diameter) per mouse were analyzed, and the number of epithelial AB-PAS positive cells per 100μm of basement membrane was quantified using Image J (NIH version 1.43).

All the tonsillar specimens were fixed in 10% formol saline for 24 h, washed, then exposed to serial concentrations of ethyl alcohol, and ended with absolute alcohol (for complete dehydration). Specimens were cleared in xylene and embedded in paraffin for 24 h. The rotary microtome prepared paraffin wax tissue blocks for sectioning at 4 μm thicknesses (LEICA RM 2125 UK). The obtained tissue sections were collected on glass slides, deparaffinized, and stained with H&E¹¹ and Sirius red^{12 (link)}. H&E stain to assess the general histological structure of palatine tonsil while Sirius red is used to assess collagen fibers deposition (fibrosis).
Slides were examined under a light microscope (Primo star, ZEISS, China). The photos were taken using (an Axiocam ERc 5 s, ZEISS, China) camera, at the Histology Department, Faculty of Medicine for Girls, Al Azhar University.

The histopathological changes in the gut of 5th instar larva were studied after 24 h exposure to sub lethal concentration of chlorpyrifos (1.5 mg/L and 2.0 mg/L). Histological experiment was performed three times in triplicates (n = 10). The gut was removed and fixed in the Carnoy's fluid (absolute alcohol: chloroform: glacial acetic acid = 6:3:1). The histological slides were prepared by the standard protocol of dehydration and paraffin embedding (Gurr, 1959 ). Sections of 5 μm were cut and stained with hematoxylene and eosin. Afterwards, the sections were observed under phase contrast microscope (Zeiss Axio Cam ERC 5S).

Intracellular accumulation assay was performed according to the protocol by Srivastava and Ahmad [5 (link)], with minor modification. Briefly, C. auris isolates were grown overnight in YPD broth medium at 37 °C. Then, C. auris suspensions were incubated with FLU and FAR as previously described in Section 2.3. Post incubation, cells were pelleted (5000× g for 5 min) and washed in sterile PBS. Cells were re-suspended in sterile PBS (1 mL) supplemented with 2% glucose and 4 μM rhodamine 6G (Sigma, Taufkirchen, Germany) and incubated at 37 °C for 30 min. Cells were then washed twice with cold sterile PBS and 1 mL of fresh PBS was added to the pellet. Afterwards, 100 μL of suspension was pipetted into a flat-bottomed dark 96 well plate (Costar^®, Kennebunk, ME, USA) and fluorescence was measured with a fluorescence spectrophotometer (Tecan, Männedorf, Switzerland). The results were evaluated using MagellanTM Data Analysis Software and the intensity of fluorescence of samples was determined by relative fluorescence units (RFUs). The same suspension was immediately used for microscopy. Fluorescence was detected by an inverted fluorescence microscope (Zeiss, Jena, Germany) with excitation/emission spectra 525/548 nm. The pictures were captured by AxioCam ERc5s (Zeiss, Jena, Germany) and evaluated by software Motic Images Plus 3 (Hong Kong, China).