Z16 apo macroscope

Quantitative fluorescent detection was performed for each experiment with Leica EL 6000, an external light source for fluorescent excitation, and with Leica Z16 APO macroscope, magnification ×0.57, with L5 Leica filter cube. This filter contains an excitation filter at 480 nm ± 20 nm, a dichroic mirror at 505 nm and a suppression filter at 527 nm ± 15 nm. These optical conditions suit well the FAM probe spectral properties (absorption wavelength: 495 nm, emission wavelength: 520 nm). Images were recorded from Hamamatsu EM-CCD Camera C900-13, one image every 10 seconds during 20 minutes, with an exposure time of 75 ms. Synchronization between shutter aperture and camera acquisition prevents from sample bleaching, and was realized thanks to EG R&D Vision delays generator. ImageJ freeware was used to normalize data by subtracting the first image to each image sequence. Signal over time is given by an average signal measurement on the test area, for each image.

Pitfall trap sampling was carried out between 11 and 18 September 2019 in ten transects, each with 20 traps set at 10 m intervals and run for six days; the traps were each filled with 50 ml of a 1:1 mixture of 95% ethanol and propylene glycol. Transects were placed in each of the main habitat types (riverine fringe, alluvial terrace gravels, alluvial fans, aeolian sands, Mispah soils and mountain slopes) within the ODM mineral lease area, which extends for approximately 16 km along the banks of the Orange River on the northern border of the Richtersveld National Park. Specimens were collected under permit # RNP03/19 issued by SANParks. Representatives of each morphospecies recognised in the samples were subsequently point-mounted for identification.
Measurements of mounted specimens were taken using a Leica MZ16 stereomicroscope equipped with an axial shift carrier and an ocular graticule calibrated against a stage micrometer. Specimens were photographed using a Leica DFC 425 digital camera connected to a Leica Z16APO Macroscope. Images were captured using Leica Application Systems (LAS) multifocus V4.9; montage images were generated using Helicon Focus V6.8.0 and edited with Adobe Photoshop CS3.

In total, 50 individuals (10 mature, 40 immature) were analysed for the presence/absence of A.M. and their gonopores were observed as well. Twelve additional individuals were taken for analysis of ontogenetic changes of gonopores. Their total body length was measured under a complanar apochromatic macroscope (Leica Z16-APO) using a millimetric paper. Individuals fixed in 70% alcohol were stained in 2% methylene blue solution and immediately observed for the location and shape of gonopores. Males were characterised by the presence of gonopores at the far proximal end of the coxa of the fifth pair of pereiopods, whilst the gonopores of females were located at the far proximal end of the third pair of pereiopods (Bauer 2004 ). In addition, mature individuals were also observed and their second pleopods were collected to detect the absence/presence of A.M. using bright-field light microscopy (Leica DMLB). Presence of an A.M. on the pleopod II corresponds to males, while its absence characterises females.
The lengths of exopodite, endopodite, basipodite and A.I. of each of the 40 immature individuals as well as their body size were measured. These individuals belong to the same age group and they have similar sizes. All measurements were made on images of the shrimps obtained by a Leica Z16-APO macroscope equipped with a computerised system of image analysis.

The specimens were preserved in 70–80% ethanol. Eight paratypes were mounted on slides with Euparal liquid. Drawings were made using a Zeiss Axioplan microscope with a camera lucida. Photographs of larvae were taken using a Leica Z16 APO macroscope and processed with Leica Application Suite™ Version 3.1.8 to obtain combined photographs with enlarged depth of field. Photographs were subsequently enhanced with Adobe Photoshop™ CS3.
Specimens used for SEM were dissected and dehydrated through a stepwise immersion in ethanol and then dried by critical point drying (Leica EM CPD300). The mounted material was coated with a 5 nm Au/Pd layer (Leica EM ACE200) and subsequently examined and photographed with a Zeiss EVO LS 15 scanning electron microscope. SEMs were subsequently enhanced with Adobe Photoshop™ CS3.

Each liver lobe was separately photographed with the Leica Z16 APO macroscope from every side. Tumor formations >1 mm in diameter were counted and measured using the calibrated Diskus software (Hilgers, Königswinter, Germany). Conventional Haematoxyline & Eosin (H&E) stainings were evaluated for signs of malignancies, and malignant areas were measured using the Diskus software in a blinded session.