Paraffin-embedded tissue was sliced, dewaxed, dehydrated, and incubated with the primary antibodies of Caspase-3 (Cas-3; E−8; sc-7272; Santa Cruz, Texas, USA) and VEGF (JH1212; sc-57496; Santa Cruz, Texas, USA), at 1/100 dilution, for 60 min. After washing with phosphate-buffered saline (PBS), the sections were incubated with biotinylated secondary antibody (EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit; ab80436; Abcam, Cambridge, UK) and streptavidin-alkaline phosphatase conjugate. The antigen dilution solution was used as a negative control instead of a primary antibody. Single blinding was performed for all analyses. Semiquantitative analyses were performed to evaluate the markers. The grading scores ranged from 0 to 3 as follows: 0 = negative, 1 = focal weak staining, 2 = diffuse weak staining, and 3 = intense diffuse staining. Ten different areas of each section were examined under 40x magnification, and the Database Manual Cell Sens Life Science Imaging Software System (Olympus Co., Tokyo, Japan) was used for microphotography.
Database manual cell sens life science imaging software system
Manufactured by Olympus
Sourced in Japan
The Database Manual Cell Sens Life Science Imaging Software System is a comprehensive software suite designed to manage and analyze data from various imaging and microscopy techniques used in life science research. The software provides a centralized database for storing, organizing, and retrieving imaging data, as well as tools for image analysis and data visualization.
Automatically generated - may contain errors
Lab products found in correlation
Hrp dab ihc detection kit micropolymer, by Abcam (2 mentions)
Expose mouse and rabbit specific hrp dab detection ihc kit, by Abcam (1 mentions)
Sc 57496, by Santa Cruz Biotechnology (1 mentions)
Leica asp300, by Leica Microsystems (1 mentions)
Leica rm 2155 rotary microtome, by Leica Microsystems (1 mentions)
6 protocols using database manual cell sens life science imaging software system
1
Immunohistochemical Evaluation of Cas-3 and VEGF
2
Immunohistochemical Analysis of Kidney Biomarkers
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All antibodies were purchased (Abcam, Cambridge, UK) and used in 1/100 dilution. Kidney samples were immunostained with primary antibodies, by PGE-2 [Anti-PGE-2 antibody (ab2318)]; C-reactive protein [Anti-C Reactive Protein antibody-Amino terminal end (ab65842)]; Anti-TNF-α antibody (ab6671)]; and Serum amyloid A [Anti-Serum Amyloid A antibody [mc1] (ab655)], according to the manufacturer’s instructions. EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit (ab80436) used as seconder kit. All the slides were analyzed for immunopositivity and a semiquantitative analysis was carried out. Samples were analyzed by examining five different sections in each sample, which were then scored from 0 to 3, according to the intensity of staining (0, absence of staining; 1, slight, 2, medium and 3, marked). Morphometric evaluation was made using the Database Manual Cell Sens Life Science Imaging Software System (Olympus Corporation, Tokyo, Japan).
3
Immunohistochemical Detection of BPV Antigens
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Sections taken on polylysined slides for the immunohistochemistry the streptavidin-biotin peroxidase complex method were used. BPV antibody [Anti-HPV antibody (BPV-1/1H8+CAMVIR) (ab2417), 1/50 dilution] was used as primary antibody. Mouse and Rabbit Speci c HRP/DAB IHC detection kit-micropolymer (ab236466) (Abcam, Cambridge, England) was used as secondary kit. For immunohistochemistry after depara nization and dehydration sections were processed according the manufacturer instruction. Sections incubated 60 min with primary antibody and antibody dilution solution was used instead of primary antibody for negative controls. Then sections dehydrated by passing through alcohol series, cleared in xylol, cover slipped and evaluated under a light microscope (Olympus CX41). Microphotography and morphometric analysis were performed using the Database Manual Cell Sens Life Science Imaging Software System (Olympus Corporation, Tokyo, Japan).
4
Immunohistochemical Detection of BPV Antigens
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Sections taken on polylysined slides for the immunohistochemistry the streptavidin-biotin peroxidase complex method were used. BPV antibody [Anti-HPV antibody (BPV-1/1H8+CAMVIR) (ab2417), 1/50 dilution] was used as primary antibody. Mouse and Rabbit Speci c HRP/DAB IHC detection kit-micropolymer (ab236466) (Abcam, Cambridge, England) was used as secondary kit. For immunohistochemistry after depara nization and dehydration sections were processed according the manufacturer instruction. Sections incubated 60 min with primary antibody and antibody dilution solution was used instead of primary antibody for negative controls. Then sections dehydrated by passing through alcohol series, cleared in xylol, cover slipped and evaluated under a light microscope (Olympus CX41). Microphotography and morphometric analysis were performed using the Database Manual Cell Sens Life Science Imaging Software System (Olympus Corporation, Tokyo, Japan).
5
Histopathological Analysis of Fish Skin Lesions
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The taxonomic names and scientific classification for fish species herein were made in accordance with FishBase (Thomson, 1990; Engin and Innal 2017; Fricke et al. 2020) .
During the necropsy, samples of fish skin lesions were collected at the site of parasitic infection. For histopathological assessment, the whole body of small fish individuals was transversally cut and fixed in 10% neutral formalin solution. Parasite attachment areas were selected and skin samples were prepared by an automatic tissue processing equipment (Leica ASP300S; Leica Microsystem, Nussloch, Germany). The tissues of fish were embedded in paraffin, and 5 μm serial sections were acquired using a Leica RM 2155 rotary microtome (Leica Microsystem, Nussloch, Germany). Subsequent, histopathological sections were stained with hematoxylin and eosin (HE) and examined under 40X magnification of a light microscope. Morphometric evaluation and microphotography were performed using the Database Manual cellSens Life Science Imaging Software System (Olympus Corporation, Tokyo, Japan).
During the necropsy, samples of fish skin lesions were collected at the site of parasitic infection. For histopathological assessment, the whole body of small fish individuals was transversally cut and fixed in 10% neutral formalin solution. Parasite attachment areas were selected and skin samples were prepared by an automatic tissue processing equipment (Leica ASP300S; Leica Microsystem, Nussloch, Germany). The tissues of fish were embedded in paraffin, and 5 μm serial sections were acquired using a Leica RM 2155 rotary microtome (Leica Microsystem, Nussloch, Germany). Subsequent, histopathological sections were stained with hematoxylin and eosin (HE) and examined under 40X magnification of a light microscope. Morphometric evaluation and microphotography were performed using the Database Manual cellSens Life Science Imaging Software System (Olympus Corporation, Tokyo, Japan).
6
Histological Analysis of Alveolar Bone Loss
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The histological lesions were examined and scored by a pathologist (Ö.Ö.) who was blinded to the groups. The left maxillary halves were fixed in formalin solution, neutral-buffered (10%) and decalcified with ethylenediaminetetraacetic acid solution (0.1 M) for 1 week. The specimens were then dehydrated through a graded ethanol series and embedded in paraffin. In order to obtain 5-µm-thick longitudinal sections, the paraffin blocks were serially cut in a mesio-distal direction along the long axis of each tooth. The specimens were stained with hematoxylin and eosin (HE). To assess the degree of ABL, the distances between the ABC and CEJ were measured. The lesioned areas were analyzed in each rat using a light microscope (Olympus CX41, Olympus Corporation, Tokyo, Japan). The Database Manual CellSens Life Science Imaging Software System (Olympus Corporation) was used for evaluation of inflammatory cell infiltration. The numbers of polymorphonuclear leukocytes (PMNLs) in the junctional epithelium and connective tissue subjacent to the epithelium and lymphocytes in the connective tissue (in a 0.05 × 0.05-mm area) were examined and counted at a magnification of ×40 (31) . The numbers of osteoclasts and osteoblasts on the lesioned alveolar bone were counted in an area 1.23 mm square (×400 magnification) (32) .
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