For histological analysis, kidneys were sliced in half transversely, fixed in 4% paraformaldehyde or formalin for 24 hours, then placed on 70% ethanol until paraffin embedding process was completed and sectioned at 5 μm. After deparaffinized in xylene and rehydrated, antigen retrieval was performed using 10 mM sodium citrate, pH 6.0, with 0.05% Tween for 25 min in pressure cooker. Sections were washed in 1X PBS 0.05% Tween, blocked using Super Block (ScyTek Laboratories, AAA125) 10 minutes and incubated with primary antibodies diluted in Normal Antibody Diluent (ScyTek Laboratories, ABB125) at 4°C overnight, and secondary antibodies for 1 hour according (Supplemental Table 2 ). Sections were then mounted using VECTASHIELD Vibrance Antifade Mounting Medium with DAPI (Vector Laboratories, H-1800), or Prolong Gold (Life Technologies, P36931). Images were obtained using an Olympus BX51 microscope and CX9000 camera (Shinjuku, Japan), or by confocal imaging using Olympus FV1000 confocal laser scanning microscope (Olympus America).
Cx9000 camera
Manufactured by Olympus
Sourced in Japan
The CX9000 is a digital camera designed for laboratory applications. It features a high-resolution sensor and advanced imaging capabilities to capture detailed samples and specimens. The core function of the CX9000 is to provide clear, accurate images for scientific and research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Super block, by ScyTek Laboratories (2 mentions)
Bx51 microscope, by Olympus (2 mentions)
Normal antibody diluent, by ScyTek Laboratories (1 mentions)
Vectashield vibrance antifade mounting medium with dapi, by Vector Laboratories (1 mentions)
Prolong gold, by Thermo Fisher Scientific (1 mentions)
Fv1000 confocal laser scanning microscope, by Olympus (1 mentions)
Kwik diff, by Thermo Fisher Scientific (1 mentions)
2 protocols using cx9000 camera
1
Histological Analysis of Kidney Tissues
2
Multicolor Immunofluorescence Staining of Cells
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50,000 cells were subject to cytocentrifugation. Slides were differentially stained (Kwik-Diff, ThermoFisher Scientific, Waltham, MA). For immunostaining, cells were permeabilized with 100% cold acetone for 2 minutes at room temperature. Slides were washed in phosphate buffered saline (PBS) with 0.05% Tween-20, treated 10 minutes with Superblock (Scytek, Logan, UT), and incubated with primary antibodies overnight at room temperature: α-RNase3 (Abcam), α-RNase6 (Cloud-Clone), α-RNase7 (Sigma), α-CD66b (Stem Cell Technologies, Vancouver, BC), α-CD68 (Stem Cell Technologies), and α-Cytokeratin (CAM5.2, Becton Dickinson, Franklin Lakes, NJ). The next day, slides were washed, hybridized with Alexafluor 488 and Alexafluor 595 conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA) for 90 minutes at room temperature, washed again, and cover slipped in mounting medium with DAPI for nuclear visualization (Vector Labs, Burlingame, CA). Slides were visualized using an Olympus BX51 microscope and CX9000 camera. Controls consisted of irrelevant primary and secondary only conditions.
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