RNA in situ hybridization was performed as described (Feng et al, 2010 (link); Prince et al, 1998 (link)). The probes used in this study: bozozok, chordin, eve1, gsc, egr2b (krox20), myoD, pax2a and squint (ndr1) were obtained from Dr. Solnica-Krezel (Washington University, Saint Louis, USA). Unless otherwise stated, images of whole mount embryos were acquired using an Olympus SZ61 dissecting microscope equipped with a high-resolution digital camera (model S97809, Olympus America) and Picture Frame 2.0 or 3.0 software (Optronics). Two-color in situ hybridization embryos were flat mounted as described and images were acquired using an AxioPlan2 microscope equipped with an AxioCam CCD camera (Zeiss). Images of fluorescent in situ hybridization embryos were acquired using Leica SP5 confocal. GFP reporter expression of the Tp1bglob:egfp line was acquired using an Olympus SZ16 fluorescent dissecting microscope and Microfire digital camera (Olympus). Images were processed in ImageJ, Adobe Photoshop, and Adobe Illustrator.
S97809
Manufactured by Olympus
The S97809 is a high-precision laboratory equipment designed for accurate measurement and analysis. It features advanced sensors and a durable construction for reliable performance in a variety of laboratory settings.
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3 protocols using s97809
1
RNA in situ Hybridization Techniques
2
Quantifying Neutrophil Infiltration in Atrial Myocardium
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The myocardial infiltration of the neutrophil cells was examined histologically. The excised lateral RA and LA tissues were fixed with 10% formalin solution and embedded in paraffin. Deparaffinized tissue sections were stained with hematoxylin-eosin. The amount of neutrophil cells in the atrial myocardium was measured microscopically. The tissue sections were observed under ×20 magnification with light microscopy (BX 51; Olympus, Melville, NY) and then directly captured with a digital camera (S97809; Olympus, Melville, NY). The captured sections were converted to image files. The intensity and extent were graded from grade 0 to 4 as previously described.18 (link) Grade 0 intensity was defined as absence of neutrophil cells in the myocardium and grade 4 was defined as dense neutrophil cell infiltration in and around the capillaries. The neutrophil cell infiltration index was calculated as the grade of the intensity × extent. All samples were graded by a histologist (KY) blinded to the experimental groups.
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Quantifying Neutrophil Infiltration in Atrial Myocardium
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