Generation of volume data by episcopic three-dimensional imaging of embryos

Episcopic fluorescence image capturing (EFIC) and high-resolution episcopic microscopy (HREM) are related techniques that are used to generate digital volume data and create three-dimensional (3D) images. Both techniques require specimens that are embedded in an appropriate medium, and images are captured from successive sections before removal from the embedded tissue block. EFIC detects autofluorescence emitted from the embedded tissue, whereas HREM requires the tissue to be stained with a fluorescent dye such as eosin. Volume data are generated as the successive sections are imaged and removed using a microtome. This protocol describes the procedure for digital data processing, visualization, and archiving for both EFIC and HREM.