Serial Registration Analysis
SERIAL REGISTRATION ANALYSIS ADD-ON
The Serial Registration Analysis add-on includes options for both serial section analysis and serial stain fusion analysis.
Serial section analysis allows users to create a tissue classifier based off a reference slide (e.g., H&E or Pan-CK) and then apply the resulting classification mask to serial section(s) stained for additional IHC markers (e.g., HER2, ER, PR, Ki-67, CD8, CD4). In the example above, a tissue biopsy section is stained with hematoxylin and eosin. A classifier has been created segmenting tumor from stroma (Tumor in red, stroma in green, glass in yellow). The serial section add-on takes the Classifier created from the H&E image and allows the user to apply the resulting classification mask to IHC-stained serial sections (e.g., ER, HER2) as regions of interest. The tumor regions, based off of the H&E classifier, are analyzed on the ER and HER2 slides using the Cytonuclear Module and Membrane Module, respectively. Serial registration analysis is compatible with both brightfield and fluorescent images and any of our HALO modules can be used for the final analysis step.
Where serial section analysis involves different tissue sections that have been stained with different markers, serial stain fusion facilitates analysis of single slides that have been stained, stripped and restained (as described in Romain Remark et al. Science Immunology, 2016). As demonstrated below, the brightfield images are deconvolved into their composite stains (nuclear counterstain and biomarker stain) and converted to a pseudo-fluorescent images in Step 1. In Step 2, the images are registered using a customizable set of configuration settings. Serial Stain Registration uses an Image Consistency Phase, a Similarity-Only Phase, and a High-Resolution Registration to precisely align single cells. Additionally, a Single Channel Registration option allows the user to align the images using one consistent fluorescent channel, typically DAPI, to improve the registration. Then, the registered serial stain images can be fused into a single composite fluorescent image that can be analyzed using Highplex FL or (any of our other FL modules).