Multimodal Staining and Imaging Protocol

H&E staining was undertaken as described previously [52 (link)]. IHC for neutrophils was undertaken as described [52 (link)] using Ly6G primary antibody (Abcam Anti-Mouse Neutrophil antibody Clone: NIMP-R14 cat. No. ab2557, Cambridge, UK) and Ly6 secondary antibody (Biocare Medical Rat on Mouse HRP Polymer cat. no. RT517L, Concord, CA USA). ApopTag staining used the Millipore ApopTag Peroxidase In Situ Apoptosis Detection kit (cat. No. S7100 Temecula, CA, USA). IHC for CHIKV capsid (monoclonal antibody 5.5G9 [54 (link)]) and ZIKA envelope (monoclonal antibody 4G2 [55 (link)], was undertaken as described using NovaRed secondary antibody (Vector Laboratories ImmPACT NovaRed Peroxidase Substrate Kit cat. No. SK-4805 Burlingame, CA, USA). Slides were digitally scanned using Aperio AT Turbo (Leica Biosystems).

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Hazlewood J.E., Dumenil T., Le T.T., Slonchak A., Kazakoff S.H., Patch A.M., Gray L.A., Howley P.M., Liu L., Hayball J.D., Yan K., Rawle D.J., Prow N.A, & Suhrbier A. (2021). Injection site vaccinology of a recombinant vaccinia-based vector reveals diverse innate immune signatures. PLoS Pathogens, 17(1), e1009215.

Publication 2021

ApopTag Peroxidase In Situ Apoptosis Detection kit ImmPACT NovaRed Peroxidase Substrate Kit Aperio AT Turbo

Anti antibody Antibody Apoptosis Capsid Clone Monoclonal antibody Mouse Neutrophil Peroxidase Polymer Vector Zika

Corresponding Organization : Australian e-Health Research Centre

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Variable analysis

independent variables

H&E staining
IHC for neutrophils using Ly6G primary antibody and Ly6 secondary antibody
ApopTag staining
IHC for CHIKV capsid using monoclonal antibody 5.5G9
IHC for ZIKA envelope using monoclonal antibody 4G2

dependent variables

Outcomes measured from the staining and IHC procedures

control variables

The experimental procedures were undertaken as described previously [52 (link)]
The secondary antibodies and detection kits used are specified (e.g., NovaRed secondary antibody, Biocare Medical Rat on Mouse HRP Polymer, Millipore ApopTag Peroxidase In Situ Apoptosis Detection kit)

positive controls

No positive controls were explicitly mentioned

negative controls

No negative controls were explicitly mentioned

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