Z-Stacking Of Single Plane Digital Widefield Fluorescent Images

By Brad Larson and Peter Banks, Applications Department, BioTek Instruments, Inc.

Incorporation of Cytation™ Cell Imaging Multi-Mode Readers and Gen5™ Data Analysis Software to Create Deconvoluted, Stacked Images of 3D Cellular Structures

Three-dimensional (3D) cellular models have the potential to become a fundamental research tool in cell biology. This is because cell culture performed in this manner re-establishes cell-cell and cell-extracellular matrix interactions that mirror what’s seen within real tissues. Histology slides, containing tissue samples mounted onto microscope slides, also continue to be an important research and diagnostic method in the clinic and laboratory. These multi-dimensional cell structures present complications for optical microscopy due to their thickness in the z-axis. Here we present a method to capture and project multiple single z-plane images using digital widefield microscopy.