Example Microscopy Metadata JSON files produced using Micro-Meta App to document the acquisition of example images using the custom-built TIRF Epifluorescence Structured Illumination Microscope

Example Microscopy Metadata JSON files produced using the Micro-Meta App documenting an example raw-image file acquired using the custom-built TIRF Epifluorescence Structured Illumination Microscope. For this use case, which is presented in Figure 5 of Rigano et al., 2021 , Micro-Meta App was utilized to document: 1) The Hardware Specifications of the custom build TIRF Epifluorescence Structured light Microscope (TESM; Navaroli et al., 2010 ) developed, built on the basis of the based on Olympus IX71 microscope stand, and owned by the Biomedical Imaging Group (http://big.umassmed.edu/) at the Program in Molecular Medicine of the University of Massachusetts Medical School. Because TESM was custom-built the most appropriate documentation level is Tier 3 ( Manufacturing/Technical Development/Full Documentation ) as specified by the 4DN-BINA-OME Microscopy Metadata model ( Hammer et al., 2021 ). The TESM Hardware Specifications are stored in: Rigano et al._Figure 5_UseCase_Biomedical Imaging Group_TESM.JSON 2) The Image Acquisition Settings that were applied to the TESM microscope for the acquisition of an example image (FSWT-6hVirus-10minFIX-stk_4-EPI.tif.ome.tif) obtained by Nicholas Vecchietti and Caterina Strambio-De-Castillia. For this image, TZM-bl human cells were infected with HIV-1 retroviral three-part vector (FSWT+PAX2+pMD2.G). Six hours post-infection cells were fixed for 10 min with 1% formaldehyde in PBS, and permeabilized. Cells were stained with mouse anti-p24 primary antibody followed by DyLight488-anti-Mouse secondary antibody, to detect HIV-1 viral Capsid. In addition, cells were counterstained using rabbit anti-Lamin B1 primary antibody followed by DyLight649-anti-Rabbit secondary antibody, to visualize the nuclear envelope and with DAPI to visualize the nuclear chromosomal DNA. The Image Acquisition Settings used to acquire the FSWT-6hVirus-10minFIX-stk_4-EPI.tif.ome.tif image are stored in: Rigano et al._Figure 5_UseCase_AS_fswt-6hvirus-10minfix-stk_4-epi.tif.JSON