Formalin-fixed paraffin-embedded myometrial (n = 5 per group), cervical (n = 5–7 per group), and chorioamniotic membrane (n = 5 per group) tissues were cut in five-μm-thick sections. Slides were deparaffinized in xylene and hydrated with decreasing concentrations of ethanol. Immunohistochemistry staining for IFNe (mouse anti-human IFNe, Clone 983338; R&D Systems) was performed using the Leica Bond Max automatic staining system (Leica Microsystems; Wetzlar, Germany). The Bond™ Polymer Refine Detection Kit (Leica Microsystems) was used to detect the chromogenic reaction of horseradish peroxidase upon oxidation of 3’3-Diaminobenzidine (DAB). Mouse IgG2B isotype (Cat. No. MAB004; R&D Systems) was used as the negative control. Brightfield images were taken using the Vectra Polaris Multispectral Imaging System and inForm software version 2.5.1. Representative images were taken at 200X magnification.
Bond max automatic staining system
Manufactured by Leica
Sourced in Germany
The Leica Bond Max is an automated staining system designed for immunohistochemistry and in situ hybridization applications. It provides consistent and reliable staining results by automating the entire staining process, from dewaxing and rehydration to counterstaining and coverslipping. The system offers a flexible and customizable staining protocol, allowing users to optimize the staining process for their specific needs.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using bond max automatic staining system
1
IFNε Expression in Reproductive Tissues
2
Immunohistochemical Analysis of CD45+ Cells
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Five-μm-thick tissue sections from mice injected with PBS or E. coli were deparaffinized and rehydrated using xylene and a series of decreasing ethanol concentrations, respectively. Immunohistochemistry staining using the Monoclonal Rabbit Anti-Mouse CD45 (AB_2799780; clone D3F8Q, cat. no. 70257S, Cell Signaling Technology, Danvers, MA, USA) was performed using the Leica Bond Max Automatic Staining System in a peroxidase-mediated oxidation of 3,3’-diaminobenzidine (DAB) from the Bond™ Polymer Refine Detection Kit (both from Leica Microsystems, Wetzlar, Germany). The negative control used was the Rabbit FLEX Universal Negative Control (cat. no. IR60066–2, Agilent, Santa Clara, CA, USA). Images were scanned using the Brightfield setting of the Vectra Polaris Multispectral Imaging System (Akoya Biosciences, Marlborough, MA, USA).
3
Immunohistochemical Analysis of SARS-CoV-2 in Placenta
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Five-μm-thick tissue sections of formalin-fixed, paraffin-embedded placental villi (PV), basal plate (BP), and the chorioamniotic membranes (CAM) were cut, mounted on SuperFrost™ Plus microscope slides (Erie Scientific LLC, Portsmouth, NH, USA), and subjected to immunohistochemistry using SARS-CoV/SARS-CoV-2 (COVID-19) spike antibody [1A9] (GeneTex, Irvine, CA, USA) and SARS-CoV-2 (COVID-19) nucleocapsid antibody (GeneTex). To serve as a positive control, tissues from pregnant women were spiked with SARS-CoV-2 (Isolate: USA/WA1/2020) (ZeptoMetrix) Culture Fluid (heat inactivated). Spiked tissues were subjected to immunohistochemistry using SARS-CoV/SARS-CoV-2 (COVID-19) spike antibody [1A9] and SARS-CoV-2 (COVID-19) nucleocapsid antibody. Staining was performed using the Leica Bond-Max automatic staining system (Leica Microsystems, Wetzlar, Germany) with the Bond Polymer Refine Detection Kit (Leica Microsystems). The mouse isotype (Agilent) and rabbit isotype (Agilent) were used as negative controls. Tissue slides were then scanned using the Vectra Polaris Multispectral Imaging System (Akoya Biosciences, Marlborough, MA, USA) and images were analyzed using the Phenochart v1.0.8 image software (Akoya Biosciences). Supplementary Table 4 summarizes the number of slides included in this study.
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