Focus On Focus: Three Technologies For Collecting 3D Image Data

Collection of 3-dimensional image data is a staple of automated microscopy. In biological science, cells often cannot be imaged in a single plane, either due to the thickness of the cell being greater than the depth of field of the microscope objective or imaging technique or to inherent unevenness across the sample. Imaging cells grown on 3D substrates is increasingly prevalent as it provides a more true-to-life representation than traditional 2D cultures. Organs or even whole animals require even greater penetration of the sample to image and render multiple cell layers or tissues. Live events happening in these samples also need to be tracked in three-dimensional space.

Materials microscopy also requires image acquisition in 3D space. Large samples, such as 12-inch semiconductor wafers, often cannot be positioned so flat that the areas of interest are always in focus when mapping the sample. Extended focus, where small Z-stacks (series of images taken at the same XY coordinates) are collected at each frame, allows an in-focus tile scan image to be stitched together at the end of the scanning routine using the best image from each stack. Multilayer samples, such as LCD screens, also require Z-stacking to image all the relevant materials therein.

This type of analysis requires the use of automated microscopy – taking 3D images by hand not only takes considerable time but also places strain on the microscope user. In contrast, motorized acquisition is repeatable and takes a matter of seconds. Prior Scientific offers three technologies to upgrade an existing microscope, or create a standalone focusing system, to allow the acquisition of 3D image data as efficiently as possible.