Paraffin sections of primary and xenograft tumors were subjected to H&E staining and immunostaining. Immunostaining was performed using the enzyme antibody method. The primary antibodies used to evaluate the epithelial area were AE1/AE3 (67306, 1:50 dilution, Cell Signaling Technology, Danvers, MA, USA), p53 (2527, 1:100 dilution; Cell Signaling Technology) due to its high expression in OC, and Ki-67 (9027, 1:200 dilution; Cell Signaling Technology) to assess cell proliferative potential. CD10 (65534, 1:100 dilution, Cell Signaling Technology) was used to evaluate the stromal cells. Each secondary antibody was specific to the primary antibody. Pathological images of the tissues were obtained using a microscope (BZ-X700 Series, Keyence, Osaka, Japan). WinROOF 2021 was used to determine the Ki-67 positivity rate.
Bz x700 series
Manufactured by Keyence
Sourced in Japan
The BZ-X700 Series is a high-performance microscope system designed for laboratory applications. It features a range of advanced capabilities, including high-resolution image capture, automated focusing, and multi-channel fluorescence imaging. The system is suitable for a variety of scientific and research purposes, providing reliable and accurate data acquisition.
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Lab products found in correlation
7 protocols using bz x700 series
1
Immunohistochemical Analysis of Tumor Samples
2
Histological Validation of PDX Models
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Tissue sections from primary and PDX tumors were prepared using standard procedures and stained with H&E. For immunohistochemical analysis, P16 (ab54210, 1:100 dilution, Abcam, Cambridge, MA, USA) and Ki-67 (M7240, 1:100 dilution, Dako Japan, Tokyo, Japan) antibodies were used to confirm that the PDX tumor was derived from the original tumor. After incubation with primary antibodies, the slides were rinsed with PBS, incubated with a species-specific secondary antibody, and stained using 3,3′-diaminobenzidine substrate solution (MBL, Osaka, Japan). Images were obtained using a microscope (BZ-X700 Series, Keyence, Osaka, Japan). The Ki-67 labeling index was determined by counting 500 tumor cells in the hotspots of nuclear labeling. Ki-67-positive cells were counted in the area with the highest proliferative activity [19 (link)].
3
Quantifying Glomerular Immunoglobulin Deposition
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Half kidneys were immediately frozen in OCT™ and subsequently sectioned at 5 µm. Following a brief acetone fixation, sections were stained with anti-IgG or anti-IgM-TexasRed (Southern Biotech, Birmingham, AL, USA) and anti-C’3-FITC antibodies (Immunology Consultants Laboratory, Inc, Portland, OR, USA). Images were obtained using the Keyence BZ-X700 Series at a fixed fluorescence intensity across slides, which was selected to minimize background fluorescence. Mean integrated fluorescence area for IgG- or IgM-TexasRed was calculated per glomerulus and averaged for each mouse. A minimum of 5 glomeruli was evaluated per mouse.
4
Immunohistochemical Analysis of Tumor Markers
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FFPE tissue sections from the patient and xenograft tumors were stained with H&E. Immunohistochemistry was performed on deparaffinized FFPE tissue sections using antibodies against ERα (ab 108398, 1:100 dilution; Abcam, Cambridge, MA, USA), PR (#8757, 1:500 dilution; Cell Signaling, Boston, MA, USA), p53 (#2527, 1:100 dilution; Cell Signaling), and Ki-67 (#9027S, 1:200 dilution; Cell Signaling). After incubation with primary antibodies, the slides were rinsed with phosphate-buffered saline (PBS), incubated with a species-specific secondary antibody, and stained using 3,3′-diaminobenzidine substrate solution (MBL, Osaka, Japan). Images were captured using a microscope (BZ-X700 Series, Keyence, Tokyo, Japan). A scale of 0–3 was used for ERα and PR (0: no staining; 1: <5% of the cells showed staining or diffuse staining that was so weak that it could not be distinguished from the background; 2: 6–50% of the cells were stained; 3: >50% of the cells were stained). Staining patterns were grouped as negative (score of 0–1) or positive (score of 2–3) [88 (link),89 (link)]. The Ki-67 index was calculated by measuring the mean percentage of Ki-67-positive nuclei among all cells in four different microscopic fields (×200) [90 (link)]
5
Histological Analysis of Tissue Samples
6
Viability and Metabolic Activity Assays
7
Live/Dead Cell Viability Assay
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Cells were cultured in well plates washed with PBS and then stained with the LIVE/DEAD viability/cytotoxicity kit (Life Technologies) by incubating with 2 μM ethidium homodimer-1 and 2 μM calcein-AM (diluted in PBS)
for 45 mins at RT in the dark. Images of cells were acquired using a Keyence fluorescence microscope (BZ-X700 Series) with 4X or 10X objectives. A metabolic activity assay was performed using the PrestoBlue cell viability reagent (Molecular Probes, Invitrogen, reference). Briefly, the old culture medium was removed and washed with PBS for each well. A fresh complete culture medium was prepared with 10% (v/v) of PrestoBlue added to the well, followed by a 60-min incubation at 37 °C and 5% CO 2 . The PrestoBlue-containing media were transferred into 96-well plates for fluorescence measurement via a plate reader (excitation at 560 nm and emission at 590 nm).
for 45 mins at RT in the dark. Images of cells were acquired using a Keyence fluorescence microscope (BZ-X700 Series) with 4X or 10X objectives. A metabolic activity assay was performed using the PrestoBlue cell viability reagent (Molecular Probes, Invitrogen, reference). Briefly, the old culture medium was removed and washed with PBS for each well. A fresh complete culture medium was prepared with 10% (v/v) of PrestoBlue added to the well, followed by a 60-min incubation at 37 °C and 5% CO 2 . The PrestoBlue-containing media were transferred into 96-well plates for fluorescence measurement via a plate reader (excitation at 560 nm and emission at 590 nm).
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