Optimized Protocol for Intestinal Swiss Rolls and Immunofluorescent Staining of Paraffin Embedded Tissue

Our research focuses on understanding how the gastrointestinal tract functions, and how these functions are impacted by different pathological states. We are interested in learning the response of different intestinal epithelial cells to various conditions, such as infection and inflammation, obesity and genetic mutations. The importance of antibody specificity is becoming increasingly apparent.

However, the impact of tissue fixation, preservation, and standardized staining methods are underappreciated in the field. Consistent and robust immunofluorescent staining greatly depends on the quality of tissue preservation, processing, and orientation that occurs in the initial steps of data collection. This protocol provides a simple and effective method for investigating various questions regarding intestinal cell biology and physiology.

Our lab is currently using these protocols to better our understanding of the roles of different intestinal cell types in maintaining homeostasis and limiting inflammation. This protocol allows detailed visualization of all regions of a mouse intestine in a single paraffin embedded block. This saves money on paraffin, and paraffin embedding services, the amount of time required for sectioning, and reagents.

It's also highly reproducible using diverse antibodies.