Immunohistochemical Analysis of Organoids

Organoids were formalin-fixed and embedded as previously described11 (link). IHC on sections of organoids and first-trimester placentas was performed using A.Menarini heat-induced epitope retrieval buffers and Vectastain avidin-biotin-HRP reagents as previously described11 (link). Primary antibodies (Supplementary Table 3) were replaced with equivalent concentrations of isotype-matched mouse or rabbit IgG for controls. Images were captured with a Zeiss Axiovert Z1 microscope and Axiovision imaging software SE64 V4.8.

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Turco M.Y., Gardner L., Kay R.G., Hamilton R.S., Prater M., Hollinshead M., McWhinnie A., Esposito L., Fernando R., Skelton H., Reimann F., Gribble F., Sharkey A., Marsh S.G., O’Rahilly S., Hemberger M., Burton G.J, & Moffett A. (2018). Trophoblast organoids as a model for maternal-fetal interactions during human placentation. Nature, 564(7735), 263-267.

Publication 2018

Axiovert Z1 microscope SE64 V4.8

Antibodies Avidin hrp Biotin Buffers Epitope Formalin Isotype Microscope Mouse Organoids Placentas Rabbit

Corresponding Organization : University of Cambridge

Other organizations : Wellcome/MRC Institute of Metabolic Science, Addenbrooke's Hospital, Wellcome Trust, Anthony Nolan, The Royal Free Hospital, University College London

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Variable analysis

independent variables

Organoids were formalin-fixed and embedded

dependent variables

IHC on sections of organoids and first-trimester placentas

control variables

Primary antibodies were replaced with equivalent concentrations of isotype-matched mouse or rabbit IgG for controls

controls

Positive control: Isotype-matched mouse or rabbit IgG
Negative control: Isotype-matched mouse or rabbit IgG

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