Journal: MethodsX

Article Title: Whole-slide imaging and a Fiji-based image analysis workflow of immunohistochemistry staining of pancreatic islets

doi: 10.1016/j.mex.2022.101856

Figure Lengend Snippet:

Article Snippet: Reagents/tools , Microscope • MetaSystems VSlide slide-scanner (MetaSystems, Altlussheim, Germany. This system consisted of: ▪ Microscope: Zeiss AxioImager Z2 microscope (Carl Zeiss Microscopy, Jena, Germany). ▪ Camera: monochrome CoolCube 1m camera (MetaSystems, Altlussheim, Germany). ▪ Stage: motorised five-position scanning stage (Marzhauser, Wetzlar, Germany). ▪ SlideFeeder x80 (MetaSystems, Altlussheim, Germany). ▪ A quad filter (filter set 81 HE; Carl Zeiss Microscopy, Jena, Germany) in conjunction with a Colibri.2 (Carl Zeiss Microscopy, Jena, Germany) light source to visualise Alexa Fluor 488, Alexa Fluor 647, and DAPI fluorophores. ▪ A filter specific to Alexa Fluor 594 (filter set 45 HQ TexasRed; Carl Zeiss Microscopy, Jena, Germany) in conjunction with an X-Cite 120PC Q (Excelitas Technologies, Waltham, Massachusetts, USA) light source to visualise the Alexa Fluor 594 fluorophore. Software: • Metafer software (version 3.12.6; MetaSystems, Altlussheim, Germany) to control light sources, filter sets, and exposure times for each assay. • VSlide software (version 1.1.107; MetaSystems, Altlussheim, Germany) to stitch raw images into one whole-slide image. • MetaViewer software (version 2.0.133; Metasystems, Altlussheim, Germany) to view stitched whole-slide images. • Fiji (version 1.53g; Fiji Is Just ImageJ, NIH, Maryland, USA) for subsequent image analysis..

Techniques: Immunohistochemistry, Imaging, Microscopy, Software, Staining, Sequencing, Generated, Sampling