Bh 2 compound microscope

The materials examined in this paper are deposited in the following institutions:
EUMJ Ehime University Museum, Matsuyama, Japan
TARI Taiwan Agricultural Research Institute, Taichung, Taiwan
SEHU Systematic Entomology, Hokkaido University, Sapporo, Japan
NMW Naturhistorisches Museum, Vienna
General observations and dissections were made under a Leica MZ95 stereomicroscope. Microstructures of the dissected parts in pure glycerin were studied under an Olympus BH-2 compound microscope. After observation, the dissected parts were mounted on the same card with the specimen. Photographs were taken under the Leica MZ95 and combined in Helicon Focus ver. 4.70.5 Pro (Helicon Soft Limited).
Morphological terminology follows Hernando and Ribera (2005) . Morphological abbreviations used in the measurements are as follows:
EL length of elytra in suture;
EW maximum width of elytra;
PL length of pronotum in median line;
PW maximum width of pronotum;
TL total length (PL+EL).
The average is given in parentheses after the range.

5 µL of fresh spore (10⁴/mL) was spotted on PDA plate with or without DHC and cultivated on 28 °C. Colony diameter was measured at the 10th day. The conidia and cleistothecia were observed using an Olympus BH2 compound microscope with differential interference contrast optics.
For biomass measurement, 1 mL of fresh 10⁶/mL spores was seeded into 50 mL of PDB containing different concentration (0, 0.5, 1.0, 2.0, 5.0, 10 mM) of DHC in 250 mL Erlenmeyer flasks with continuous shaking at 150 rpm at 28 °C for 10 days (three replicates per concentration). Then fermented broths were centrifuged for 5 min at 2200g. The precipitation was washed three times with distilled water and dried to a constant weight at 45 °C as biomass, and the supernatant was collected to detect mycotoxin (citrinin).

Kernels were removed from the FAA fixative, dehydrated through a graded tertiary butyl alcohol series, and embedded in Para last Plus wax (Sigma-Aldrich, St. Louis, MO, United States, Cat. #P3683) (Berlyn and Miksche, 1976 ). Tissue sections (10 μm) were cut longitudinally from the wax-embedded kernels using an American Optical Rotary microtome, affixed to glass microscope slides with Haupt’s gelatin adhesive, and stained with 0.05% aqueous toluidine blue. Cover slips were cemented over stained sections using a Permount mounting media (Fisher Scientific, Fair Lawn, NJ, United States, Cat. #SP15-100), and viewed with an Olympus BH-2 compound microscope.

All the specimens from hosts collected for this study and selected for morphological examination were cleared in lactophenol and viewed as temporary wet mounts using an Olympus BH-2 compound microscope. Light micrographs were taken using the same microscope. Measurements in micrometres were taken with the aid of an ocular micrometer and are given as the range where more than two measurements were taken. En face and transverse sections of the anterior body of representative specimens were prepared using a cataract scalpel and mounted in polyvinyl lactophenol for examination. All such specimens examined morphologically for this study, except those used for sectioning, were deposited in the South Australian Museum (SAMA) Adelaide, South Australia (Voucher numbers for specimens from Senegal are AHC 48827–48830, and all others AHC 48791–48826 and AHC 48831–48837).

Specimens of Macuxitermescolombicus sp. n. were collected in Departamento Magdalena, Colombia, on 3 JUN 2009. Images of preserved specimens in 85% ethanol were made using an Olympus SZX9 stereomicroscope fitted with a LM Scope camera tube PageBreakto an Olympus E-410 digital camera. Specimens were suspended in Purell® Instant Hand Sanitizer for transparent posturing support during photography. Enteric valve slide images were taken with an Olympus BH-2 compound microscope fitted with phase contrast optics. The entire worker P2 region was removed by micro-dissection and external muscle detached. Food particles were removed from enteric valve armature using an ultrasonic cleaner. The cleaned enteric valve was longitudinally cut, splayed open, and mounted on a microscope slide using PVA medium (BioQuip Products Inc.). External morphological terminology follows that of Roonwal (1969) and internal anatomical terminology that of Noirot (2001) .

Spores of Adiantum species were obtained for measurement by tapping the abaxial side of sporulating tissue onto a microscope slide under a dissecting scope and gently crushing the sori to release spores if necessary. For the western North American Polystichum sample, spores were harvested by inserting a probe coated with Hoyer's medium into a sorus with dehisced sporangia, then dispersing the harvested material into Hoyer's medium on a microscope slide. All spores were visualized using an Olympus BH‐2 compound microscope (Olympus Corporation, Center Valley, Pennsylvania, USA) at 400× or 600× magnification and photographed with a smartphone camera (LG V30, LG Electronics, Seoul, South Korea) and mount system (Gosky Universal Cell Phone Adapter Mount, Gosky Optics, Atlanta, Georgia, USA). Spores with irregular shape were excluded from measurement. For Adiantum, irregular spores were diagnosed by (1) absence or irregularity of their sporopollenin coat, making them transparent to some degree, and (2) by a drastic departure from the typical trilete shape. For Polystichum, spores were excluded as irregular if they were not ellipsoidal.
Spore measurements were performed in ImageJ (Schneider et al., 2012). Calibration was performed using an ocular micrometer calibrated with an object micrometer.

The specimens examined for this study are lodged in the Denver Museum of Nature & Science, Colorado (DMNS) and the Western Australian Museum, Perth (WAM). They were studied using temporary slide mounts prepared by immersion of the specimens in lactic acid at room temperature for several hours, and mounting them on microscope slides with a 10 mm coverslip supported by small sections of 0.25 mm diameter nylon fishing line. After the study, the specimens were rinsed in water and returned to 75% ethanol with the dissected portions placed in 12 × 3 mm glass genitalia microvials (BioQuip Products, Inc.). The specimens were examined with a Leica MZ16 A dissecting microscope and an Olympus BH2 compound microscope, and illustrated with the aid of a drawing tube attached to the compound microscope. Measurements were taken at the highest possible magnification using an ocular graticule.
Terminology and mensuration mostly follow Chamberlin (1931) , with the exception of the nomenclature of the pedipalps, legs and some minor modifications to the terminology of the trichobothria (Harvey 1992 (link)), chelicera (Judson 2007 (link)) and faces of the appendages (Harvey et al. 2012 (link)).