The hepatopancreas and muscle tissues of shrimps challenged with V. parahaemolyticus (with or without 24 h incubation at 6 g/L of P. tectorius leaf extract) and control shrimps (without leaf extract and Vibrio exposure) were fixed in Davison reagent for 48 h. Tissues were placed in 70% alcohol for 30 min before being transferred to a cassette and then to an automatic tissue processing machine (Leica™ 1020, Germany). The samples were then placed on a steel cassette and embedded in paraffin (melting point 54–56 °C). The paraffin blocks were allowed to freeze on a cold plate (Leica™ EG1150C, Germany) and then refrigerated overnight at 4 °C. The blocks were kept cool on a cold plate before being sectioned with an automated microtome (Leica™ RM2255 Microtome, Germany) at a thickness of 5 μm. Before staining with Harris hematoxylin and eosin, sample sections were mounted on albumin-coated glass slides (Sigma, USA). Slides were fixed with dibutylphthalate polystyrene xylene (DPX), and histological images were examined with a Leica™ DM LB2 Light Microscope (Germany) at 40x magnification and a Leica™ Image Analyzer System (Germany).
Image analyzer system
Manufactured by Leica
Sourced in Germany
The Image Analyzer System is a versatile laboratory equipment designed for high-quality digital image acquisition and analysis. It provides a robust and reliable platform for a wide range of imaging applications in various research and industrial settings.
Automatically generated - may contain errors
3 protocols using image analyzer system
1
Shrimp Histopathology with Vibrio and Leaf Extract
2
Pancreatic Islet Histological Analysis
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Upon dissection, livers, kidneys, and pancreases were removed from the carcasses. Samples were fixed with 10% buffered formalin for 24 hours before the dehydration process using a tissue processing machine. Then, samples were embedded into molds with paraffin wax and sectioned at 5 μm. The islet areas were measured via selecting the six largest pancreatic islets/group in all groups using a Leica™ DM LB2 light microscope (Germany) that was equipped with the Leica™ Image Analyzer System. The LAS 4.0 system software and a ×20 objective lens were used to measure the area (μm2) of the islets of Langerhans.
3
Quantifying Osteoclast Activity via TRAP
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Tissue sections were deparaffinised and TRAP staining was performed using a commercial acid phosphatase leucocyte kit (Sigma, St Louis, MO). In the bone samples, ten areas (magnification × 20) were randomly observed with an optic microscope (Olympus-BX51, Germany) connected to an image analyzer system (Leica-Qwin, UK) and TRAP positive cells in each area were observed.
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