After organ culture for 7 days, thoracic aorta segments were washed with PBS and fixed with 10% neutral buffer formalin. The specimens were then embedded in paraffin, and 5μm cross-sections were cut. The sections were deparaffinized and H&E staining was performed. Arterial medial calcification was visualized using Alizarin Red S staining as previously described 23 (link). For immunohistochemistry, the sections were deparaffinized, followed by treatment with citrate buffer for antigen retrieval and 3% H2O2. The sections were blocked with Dako serum-free blocking solution (Dako North America, USA) and incubated with primary antibody for overnight at 4°C. The primary antibodies included SM22α (Abcam, ab14106), Osterix (Abcam, ab94744). Subsequently, the sections were incubated with biotinylated secondary antibodies for 30 min at room temperature. Avidin-biotinylated enzyme complex (Vector Laboratories, USA) and a diaminobenzidine substrate chromogen system (Dako North America, USA) were used for detection. Sections were counterstained with hematoxylin. The images were captured by Olympus DP260.
Dp260
Manufactured by Olympus
Sourced in Japan
The DP260 is a digital camera module designed for microscope systems. It features a high-resolution CMOS sensor and supports live image capture, recording, and image analysis capabilities.
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Lab products found in correlation
2 protocols using dp260
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Histological Analysis of Aortic Calcification
2
Procedure for Detecting Phorticidae Parasites
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First-stage larvae were squeezed out of the mature female worms and placed on a slide with a drop of saline solution and observed under the light microscope (OLYMPUS DP260, Japan).
L1 were transferred to a concave slide with several drops of water after collection, and three-day-old, fermented pear juice (water-pear juice 1:1). Mature P. okadai (n = 100, F:M = 1:1), with food and water restricted 4 h prior, were used for experimental infection. The slide with L1 was placed in well-sealed cages (22 × 22 × 27 cm). Every 20 min, 1 mL of the medium was added to attract P. okadai to the feed. This process lasted for 2 h, after which the flies were normally fed (Supplementary Materials S1: Video S1: Chapter S3 ).
Otranto et al. reported a procedure in which live and dead P. okadai were randomly collected every two days, examined via dissection, and subjected to molecular analysis with cox1 [21 (link)], as described above, until a positive infection was detected (Figure 3 ). Initially, the proboscis of the flies was stretched to detect positive L3, followed by dissection of the head, thorax, and abdomen with the purpose of detecting the presence or absence of other developmental stages of T. callipaeda.
L1 were transferred to a concave slide with several drops of water after collection, and three-day-old, fermented pear juice (water-pear juice 1:1). Mature P. okadai (n = 100, F:M = 1:1), with food and water restricted 4 h prior, were used for experimental infection. The slide with L1 was placed in well-sealed cages (22 × 22 × 27 cm). Every 20 min, 1 mL of the medium was added to attract P. okadai to the feed. This process lasted for 2 h, after which the flies were normally fed (
Otranto et al. reported a procedure in which live and dead P. okadai were randomly collected every two days, examined via dissection, and subjected to molecular analysis with cox1 [21 (link)], as described above, until a positive infection was detected (
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