Tissue sections (5-μm) were baked at 60 °C for an hour and deparaffinized with xylene and a graded series of ethanol. Antigen retrieval was conducted using a Decloacking chamber (Biocare Medical) at 90 °C for 15 min in AR6 buffer (Akoya Biosciences). Multiplex fluorescent immunostaining was conducted following the Opal four-color user manual (Akoya Biosciences), including a nuclear 4′,6-diamidino-2-phenylindole (DAPI) counterstain. A tracheobronchial lymph node from a cat determined to no longer have systemic infection was used simultaneously as a negative control for GFP and a positive control for myeloid/histiocytic antigen and CD20. Whole-slide images were acquired using a Zeiss Axio Scan Z.1 whole-slide scanner at 200× equipped with a Colibri 7 LED light source and 16-bit Orcha Flash 4.0 monochrome camera. Immunohistochemical and acquisition parameters are outlined in SI Appendix, Table S3 .
Decloacking chamber
Manufactured by Biocare Medical
The Decloacking Chamber is a specialized laboratory device designed to prepare and condition samples for analysis. It serves the core function of exposing samples to controlled temperature and pressure conditions, enabling the effective retrieval of target analytes or markers from the sample material.
Automatically generated - may contain errors
Lab products found in correlation
Axio scan z1, by Zeiss (1 mentions)
Ar6 buffer, by Akoya Biosciences (1 mentions)
Immpact novared peroxidase substrate, by Vector Laboratories (1 mentions)
Antigen unmasking solution, by Vector Laboratories (1 mentions)
Da vinci green diluent, by Biocare Medical (1 mentions)
Vectastain abc kit, by Vector Laboratories (1 mentions)
2 protocols using decloacking chamber
1
Multiplex Immunofluorescence Staining of Tissue Sections
2
Immunohistochemical Analysis of CLCA1 in Mouse Kidneys
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Mouse kidneys were fixed in 10% formaldehyde and embedded in paraffin, and 4‐μm sections were cut. Sections were incubated with Xylene‐S (Cat. # HC‐700, Thermo Fisher Scientific) to remove paraffin and washed by ethanol. After deparaffinization, the slides were incubated with antigen unmasking solution (Cat. #, H3300, Vector Laboratories) at 125°C for 30 s and 90°C for 10 s using a decloacking chamber (Cat. #, DC2002, Biocare Medical LLC). The staining was performed by Vectastain ABC kit protocol (Cat. #, PK‐4001, Vector Laboratories). CLCA1 antibodies were diluted (1:50) in Davinci Green diluent (Cat. #, PD‐900L, Biocare Medical). After incubation with Vectastain ABC kit, the slides were incubated with ImmPACT NovaRED peroxidase substrate (Cat. #, SK‐4805, Vector Laboratories) for 3–4 min at RT followed by hematoxylin staining for 2 min at RT.
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