Transgenic fish were generated by microinjection of test constructs together with 50–100 pg of Tol2 transposase mRNA into fertilized 1-cell-stage embryos [42 (link)]. Three day-post-fertilization larvae were screened for mCherry fluorescence in the majority of heart cells. Larvae were anesthetized and larval fin folds were amputated at a position immediately caudal to the end of the notochord using a 22.5 degree microknife (FST) and PDMS cutting surface. Fish were then embedded in 1.5% low-melt agarose and imaged on Zeiss LSM 7 microscope with the Plan-Neofluar 10× (numerical aperture 0.30) objective. Time-lapse images were captured at a frequency of 1 z-stack every 10, 7, or 5 min for up to 12 h. Adult fins were amputated using a razor blade approximately halfway along the length of the fin and maintained at 28 °C. Adult fish were imaged on either a Nikon SMZ1500 outfitted with the Nightsea Stereomicroscope Fluorescence Adapter system and imaged with a Nikon D80 camera or on a Zeiss Stemi SV11 Apo and imaged with a Zeiss AxioCam HRc camera.
Stemi sv 11 apo
Manufactured by Zeiss
Sourced in Germany
The Stemi SV 11 Apo is a stereo microscope designed for high-resolution imaging and analysis. It features an apochromatic optical system for superior image quality and magnification ranging from 6.3x to 80x. The microscope is equipped with a trinocular observation tube, allowing for the connection of a camera or other imaging device.
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Lab products found in correlation
Eclipse e800, by Nikon (2 mentions)
Eos rebel t3i camera, by Canon (2 mentions)
Axiovert 25, by Zeiss (2 mentions)
Axiocam mrc5, by Zeiss (2 mentions)
Axiocam hrc camera, by Zeiss (1 mentions)
Lsm 7 microscope, by Zeiss (1 mentions)
Polaron sc 7620 sputter coater, by Quorum Technologies (1 mentions)
Leo 1430, by Zeiss (1 mentions)
10 protocols using stemi sv 11 apo
1
Transgenic Fish Fin Regeneration
2
Fracture Analysis of Metal-Porcelain Samples
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After the 3-point bending test, the surfaces of the fractured specimens were evaluated using a stereomicroscope (Stemi SV 11 APO, Carl Zeiss, Oberkochen, Germany). Failure types of specimens were recorded at this stage. The metal-porcelain bonding area of samples randomly selected from 8 groups has been examined with SEM (ZEISS LEO 1430, Carl Zeiss, Oberkochen, Germany) under ×1000 magnifications. Before evaluation of SEM images, specimen surfaces were sputtered with an Au-layer (Polaron SC 7620 Sputter Coater, Quorum Technologies Ltd., Kent, GB).
3
Wheat Microbiome under Heavy Metal Stress
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Healthy seeds of wheat variety Bhakkar-2000 (KJ672075) were surface sterilized with 70% ethanol for 1 min followed by 1% perchloric acid for 30 sec and rinsed thrice with ddH2O. The seeds were then incubated to germinate for 24 h at 28 °C in petri plates. To investigate the wheat plants association with microbes under heavey metal stress, the germinated seeds from petri plates were transferd to autoclaved pots containing 3 kg soil and grown for 4 weeks in greenhouse with day light of 12 h and temperature of 22–35 °C. The experiment was carried out in triplicate, each replicate comprised of 10 pots and each pot contained 6 seedlings (total = 6 × 10 × 3 = 180 seedlings per treatment). After one week of growth, the bioactive strain (metal-resistant) grown in Czapek broth (20 ml containing 3 g of fungal mycellium) was applied to the plants according to the Khan and Lee [35 (link)]. After 15 days of growth, 80 ml of waste water collected from industrial sites of Gadun was applied to the plants for 2 weeks at 48 h of interval. The plants were finally harvested after 4 weeks for subsequent analysis. Endophyte non-inoculated wheat plants under similar conditions and setup were used as a control. To see endophytic fungal colonization potential in plant tissues, 0.5–8 mm slices of wheat roots and shoots were visualized using light microscope (Stemi SV 11 Apo, Carl Zeiss).
4
Endophytic Fungal Identification Protocol
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The endophytic fungal culture was morphologically identified based on their growth characteristics, included color and shape, mycelium type and conidiophore structure. Microscopically the strains spore, hyphae and reproductive structures were identified through lacto phenol staining technique by light microscope (Stemi SV 11 Apo, Carl Zeiss) by the method of Nagamani et al., [39 ] Macroscopic photographs were taken by digital camera (Canon Co., Ltd, Japan).
5
Meiofauna and Macrofauna Abundance Analysis
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The abundance of the meiofauna (e.g., nematodes, rotifers and crustaceans) and macrofauna (different dipteral larvae and coleopteran larvae) was determined under a stereomicroscope (Zeiss Stemi SV11 Apo, Jena, Germany) at 40× magnification without sieving. The organic material containing minute invertebrates (meiofauna) was preserved in a 4 % formaldehyde solution and stained with 1 % Rose Bengal. At least 50 nematodes were mounted on slides following Seinhorst [33 (link)] and subsequently identified whenever possible to species level under a Leitz Dialux microscope (1250×) with differential interference contrast. We used standard identification keys (e.g., [34 –36 ]) and the listed references for species and genera in these books.
Nematode species were assigned to feeding-types (deposit-feeders, epistrate-feeders, suction-feeders and chewers) based on the morphology of their buccal cavity and pharyngeal structure [37 ]. Deposit-feeding nematodes show an unarmed buccal cavity, only enabling them to ingest particles in the bacterial size-range. Epistrate-feeders possess a small tooth mainly feeding on algae. In contrast, larger suction-feeding or chewing nematodes possess a stylet or large sclerotized teeth, enabling them to prey on a wider range of food items, including invertebrates that are larger than themselves.
Nematode species were assigned to feeding-types (deposit-feeders, epistrate-feeders, suction-feeders and chewers) based on the morphology of their buccal cavity and pharyngeal structure [37 ]. Deposit-feeding nematodes show an unarmed buccal cavity, only enabling them to ingest particles in the bacterial size-range. Epistrate-feeders possess a small tooth mainly feeding on algae. In contrast, larger suction-feeding or chewing nematodes possess a stylet or large sclerotized teeth, enabling them to prey on a wider range of food items, including invertebrates that are larger than themselves.
6
Microscopy Imaging Protocol for Tissue Samples
7
Macro-Morphological Characterization of Plant Species
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Photographs of the plant habit and macro-morphological characters were taken in the field. Morphological observations and measurements of the new species were conducted based on living and dried specimens and preserved materials. All morphological characters were observed and photographed with a Zeiss Stemi SV 11 Apo stereoscopic microscope and a Zeiss AxioCam MRc 5 microscope camera. Seed coat characters were examined by a Hitachi S-3400N scanning electronic microscope.
8
Morphological Analysis of New Plant Species
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Photographs of the habit and macro-morphological characters were taken in the field. Morphological observations and measurements of the new species, based on living PageBreakand dry plant specimens and preserved materials, were carried out. All morphological characters were observed and photographed with a Zeiss Stemi SV 11 Apo stereoscopic microscope and a Zeiss AxioCam MRc 5 microscope camera. Seed coat characters were revealed by a Hitachi S-3400N scanning electronic microscope.
9
Microscopy Imaging Protocol for Tissue Samples
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Images of histological sections were taken on a Nikon Eclipse E800 scope with Plan Fluor Apo objectives (magnification/numerical aperture: 4x/0.13, 10x/0.45, 20x/0.75). For picrosirius red, a polarizer was used at 90° relative to incident light. Images for adipogenesis and osteogenesis were taken on a Zeiss Axiovert25 scope equipped with 20x/0.3 phase objective. Images for chondrogenesis were taken on a Zeiss Stemi SV 11 Apo scope equipped with a S1.0x objective. For these image acquisitions, Canon EOS Rebel T3i cameras and EOS Utility software were used.
10
Mandible Harvesting and Imaging Protocol
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Under general anesthesia with isoflurane and after the subcutaneous administration of 0.05 mg/kg buprenorphine, intracardiac perfusions were performed with barium sulphate as described in Blery et al.7 (link). At the end of the procedure, mandibles were harvested, pictures of the explanted mandibles were taken using binocular imaging (Stemi SV11 Apo, Zeiss, Oberkochen, Germany). Mandibles were fixed in a solution of 4% formaldehyde over 7 days, then treated with papain for 16 h to remove as much flesh as possible and finally stored in 70% ethanol. For µCT acquisitions, as much soft tissue as possible was carefully removed and the left and right mandibles were separated.
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