Using Robotic Systems to Process and Embed Colonic Murine Samples for Histological Analyses

Abstract

The understanding of human diseases has been greatly expanded thanks to the study of animal models. Nonetheless, histopathological evaluation of experimental models needs to be as rigorous as that applied for human samples. Indeed, drawing reliable and accurate conclusions is critically influenced by the quality of tissue section preparation. Here, we describe a protocol for histopathological analysis of murine tissues that implements several automated steps during the procedure, from the initial preparation to the paraffin embedding of the murine samples. The reduction of methodological variables through rigorous protocol standardization from automated procedures contributes to increased overall reliability of murine pathological analysis. Specifically, this protocol describes the utilization of automated processing and embedding robotic systems, routinely used for the tissue processing and paraffin embedding of human samples, to process murine specimens of intestinal inflammation. We conclude that the reliability of histopathological examination of murine tissues is significantly increased upon introduction of standardized and automated techniques.

Effect of Sishen Pill on Memory T Cells From Experimental Colitis Induced by Dextran Sulfate Sodium

“…The colonic tissues were collected and fixed in 4% polyformaldehyde solution for 7 days and then dehydrated and embedded in paraffin. Five-micrometer-thick sections were cut and stained with hematoxylin and eosin (H&E), as described in previous study ( Cribiu et al., 2018 ; Cribiu et al., 2019 ) and observed under an Olympus microscope (scanning resolution 50,000 pixels per inch, 0.5 µm per pixel with 10× objective and 2.5 µm per pixel when scanning at 4×). Histopathological evaluation of colonic tissue (n = 10) was performed according to the standards reported by Mahler et al.…”

Fecal Microbiota Transplantation Controls Murine Chronic Intestinal Inflammation by Modulating Immune Cell Functions and Gut Microbiota Composition

“…Inclusion of murine tissue samples was performed with a robotic system (SAKURA Tissue-Tek) as previously described [29,30]. Hematoxylin and Eosin staining was performed as described in [29,30] and snapshots of tissue histology were taken using an Aperio CS2 microscope (scanning resolution 50,000 pixels per inch, 0.5 µm per pixel with 10× objective and 2.5 µm per pixel when scanning at 4×). Disease activity scoring was performed according to the evaluation of the criteria listed in Supplementary Table S1.…”