Fish were collected from the two great Nicaraguan lakes, Managua and Nicaragua, and five crater lakes (Asososca León, Apoyeque, Xiloá, Masaya and Apoyo) during the months of November–December of three consecutive years (2017–2019). This period represents the end of the rainy season, and the time when fish breed, which allows for clear identification of fish to species level due to breeding coloration. Fish were caught with gill nets, anaesthetized with tricaine mesylate (MS-222) and photographed on a lateral standardized position for species identification. Fish were euthanized in cold water and immediately examined; fin clips were preserved in ethanol. Euthanized fish were screened for ecto- and endoparasites using a Leica EZ4 stereomicroscope. First, the fish external surface (skin, eyes, and mouth) was observed. Then, fish were dissected, and internal organs (gut, mesentery, hearth, muscle and gall, swim, and urinary bladders) were analyzed. All recovered parasites were rinsed in saline solution and stored in 100% ethanol. Representative specimens of each parasite taxa were fixed in nearly boiling 4% formalin for morphological analysis. Some specimens were fixed in 100% ethanol for DNA extraction. Gill arches were dissected and stored in 100% ethanol. Later, in the laboratory, the gills were screened for ectoparasites using a Leica EZ4 stereomicroscope.
Ez4 stereomicroscope
Manufactured by Leica
Sourced in Germany, China
The EZ4 stereomicroscope is a high-quality optical instrument designed for a variety of applications. It features binocular observation, a magnification range, and a LED illumination system. The EZ4 provides a clear and detailed view of specimens under examination.
Automatically generated - may contain errors
Lab products found in correlation
Matlab, by MathWorks (1 mentions)
Eosin y, by Merck Group (1 mentions)
Vhx 5000 digital microscope, by Keyence (1 mentions)
Eclipse ni compound microscope, by Canon (1 mentions)
Avertin, by Merck Group (1 mentions)
Rebelt5i, by Canon (1 mentions)
Eclipse 80i, by Nikon (1 mentions)
Xylol, by Merck Group (1 mentions)
21 protocols using ez4 stereomicroscope
1
Parasites of Nicaraguan Lake Fishes
2
Ice Crystal Formation in Hydrogels
3
Wing Staining of Fly Species
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According to our recently published protocol (Wang et al., 2016 ), ten to twenty flies of each sex and species were incubated for 20 min with Eosin Y (0.5%, Sigma-Aldrich) at different temperatures. After staining, flies were washed with tap water at room temperature. Wings were cut off using micro-scissors, observed under a Leica EZ4 stereomicroscope and imaged using the in-built Leica camera and software.
4
Root Development Quantification Protocol
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To quantify root development, root samples were harvested three weeks post-inoculation with R. irregularis, rinsed with tap water, and subsequently preserved in 50% (v/v) ethanol as described previously.51 (link) The length of the primary root was determined using a ruler, and the first-order of lateral roots were counted using a Leica EZ4 stereomicroscope.
5
Quantifying Mycorrhizal Colonization in M. truncatula
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M. truncatula wild-type and mutant plants were cultivated in 48-well trays containing a mixture of soil substrates (3:3:1 mixture of Turface MVP, play sand, and LC1 grower mix). They were inoculated with Rhizophagus irregularis at a rate of 100 spores per plant for low-concentration inoculation and 300 spores per plant for high-concentration inoculation. For Gigaspora margarita inoculation, an inoculation strength of 5 % (v/v) per plant was used. For the co-cultivation of lyk8/cerk1 plants with nurse plants, one wild-type M. truncatula R108 was grown alongside one lyk8/cerk1 plant. Roots colonized by mycorrhizae were harvested at specified time points and subjected to staining with ink or 0.05% Trypan Blue. To assess the extent of mycorrhizal colonization, we used the gridline intersect method.71 (link) Briefly, root segments were cut into smaller pieces and randomly distributed across a square Petri dish, which featured a 1 cm × 1 cm grid on its base. Using a Leica EZ4 stereomicroscope, 100 grid intersections were examined for each root sample to evaluate the AMF colonization and infection events. Representative images were taken using a Keyence VHX-5000 Digital Microscope (Keyence, UK).
6
Anatomical Measurements and Histological Analysis of Molluscan Samples
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The samples obtained were transported to the laboratory following the recommendations of Barrientos (2003) . The specimens were relaxed and sacrificed by placing them in airtight containers with water and Parafarm® menthol crystals for 12–16 h. The separation of soft tissue was performed under a Leica EZ4 stereo microscope. The total length of the shell was measured with a millimetric vernier caliper (precision of 0.01 mm). After the dissection and anatomical observation, the penile complex was measured under the stereo microscope with a micrometer eyepiece from the base of the penile sheath to the end of the flagellum.
For histological analysis, the gonad was fixed in Raillet-Henry’s solution for 12 h and preserved in 70% (v/v) aqueous ethanol followed by dehydration in an increasing sequence of alcohols. For inclusion, the tissue was immersed in Histoplast®, slices of 7 µm were made with a Minot-type microtome and stained with Mayer’s hematoxylin-eosin before mounting in synthetic balsam. The histological preparations were observed and photographed under a Leica-DMLS light microscope.
For histological analysis, the gonad was fixed in Raillet-Henry’s solution for 12 h and preserved in 70% (v/v) aqueous ethanol followed by dehydration in an increasing sequence of alcohols. For inclusion, the tissue was immersed in Histoplast®, slices of 7 µm were made with a Minot-type microtome and stained with Mayer’s hematoxylin-eosin before mounting in synthetic balsam. The histological preparations were observed and photographed under a Leica-DMLS light microscope.
7
Fungal Diversity in Thai Forest Litter
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Wood litter samples were collected from forest areas in Chiang Rai and Chiang Mai, Thailand. Morphological studies were performed following the methods described by Senanayake et al. (2020) (link). The fungal structures were examined using a Leica EZ4 stereomicroscope. The micro-morphological features were observed and photographed using a Nikon ECLIPSE Ni compound microscope with a Canon 600 D digital camera. The Tarosoft Image Frame Work programme was used to measure specimen structures, and photo plates were prepared using the open-source Inkscape v.1.3 (https://inkscape.org/ ).
Pure cultures were obtained through single spore isolation on Difco potato dextrose agar (PDA ) using the spore suspension method (Choi et al. 1999 ). Germinating spores were transferred to a new PDA plate and incubated at room temperature for seven days. Ex-type pure living cultures were deposited in the
Mae Fah Luang University Culture Collection (MFLUCC ) and herbarium material was deposited in the
Mae Fah Luang University Fungarium (MFLU ), Chiang Rai, Thailand. Faces of fungi numbers (FoF ) (Jayasiri et al. 2015 (link)) and Index Fungorum numbers (Index Fungorum 2023 ) were obtained as instructed and the data were uploaded to the Greater Mekong Subregion in the GMS database (Chaiwan et al. 2021 (link)).
Pure cultures were obtained through single spore isolation on Difco potato dextrose agar (
8
Monitoring Insect Life Cycle Parameters
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Every four days (throughout 36 days that the experiment lasted), 10 tubes from each rearing chamber temperature were checked. The artificial medium was carefully dissected (destructive monitoring) and the number of eggs, larvae, pupae and adults (males and females; Supplementary Fig. S2 online) was counted. In addition, for this last stage of development, living and dead individuals were recorded. Observations were made with a Leica EZ4 stereomicroscope. The experiment took place over 36 days, totaling nine colony monitoring openings. Individual counts at each colony opening date were averaged and accumulated. With this final accumulated average of the experiment for each stage of development and temperature, the biological parameters of the species were estimated.
9
Survival Curve Establishment for eed+/- Zebrafish
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For the establishment of survival curves, the tip of the caudal fin of embryos from an eed+/− incross was transected within the pigment gap distal to the circulating blood for genotyping purposes [46 (link)]. The embryos were then pooled according to their genotypes, placed into separate 1-L tanks, and incubated at 28 °C. Larvae were fed from 5 dpf on, 3 times per day, with early-stage zebrafish nutrition (Gemma Micro ZF75, Planktovie, France). The larvae were regularly examined directly in the tanks using a Leica EZ4 stereomicroscope with no or minimal manipulation during a period of 18 days after the spawn. Larvae were declared dead when heartbeats could not be detected. Dead larvae were immediately removed from the tanks.
10
Chronic Constriction Injury Protocol
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For the CCI model, mice were anesthetized with avertin (0.4–0.6 mg/gm, i.p., Sigma-Aldrich, Saint Louis, MO, USA) and the right sciatic nerve was exposed at the mid-thigh level. Three ligatures (3-0, silk, Unik, Taiwan) were tied loosely around the nerve at about 1 mm apart. In sham-operated mice, the same procedure was performed but without ligating the nerve.
After surgery, mice underwent behavioral tests or were killed at specified weeks to collect the sciatic nerve or DRG. The sciatic nerve was excised and observed under a dissecting microscope (Leica EZ-4 stereo microscope, Bensheim, Germany) or fixed for histological staining. In some experiments, lumbar 4, 5 DRG were isolated after CCI surgery for measuring gene expression or immunostaining.
After surgery, mice underwent behavioral tests or were killed at specified weeks to collect the sciatic nerve or DRG. The sciatic nerve was excised and observed under a dissecting microscope (Leica EZ-4 stereo microscope, Bensheim, Germany) or fixed for histological staining. In some experiments, lumbar 4, 5 DRG were isolated after CCI surgery for measuring gene expression or immunostaining.
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