Intestinal Morphometric Analysis Protocol

The procedure for intestinal morphometric analysis was as described by Oladokun et al. [48 (link)]. Briefly, fixed intestinal tissues were embedded in paraffin, sectioned (0.5 μm thick), and stained with hematoxylin and eosin for morphological examinations. In each cross-sectioned tissue, ten morphometric measurements including the villus height (from the base of the intestinal mucosa to the tip of the villus excluding the intestinal crypt), villus width (halfway between the base and the tip), crypt depth (from the base upward to the region of transition between the crypt and villi) [53 (link)] per slide were carried out using Leica 1CC50 W microscope at 4× Magnification (Leica Microsystems, Wetzlay, Germany) and an image processing and analysis system (Leica Application Suite, Version 3.4.0, Leica Microsystems, Wetzlay, Germany). The total mucosa thickness (villus height + crypt depth) was subsequently calculated from the obtained data.

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Oladokun S., MacIsaac J., Rathgeber B, & Adewole D. (2021). Essential Oil Delivery Route: Effect on Broiler Chicken’s Growth Performance, Blood Biochemistry, Intestinal Morphology, Immune, and Antioxidant Status. Animals : an Open Access Journal from MDPI, 11(12), 3386.

Publication 2021

Leica Application Suite, Version 3.4.0

Crypt Embedded paraffin Eosin Examinations Intestinal Intestinal mucosa Microscope Mucosa Tissue

Corresponding Organization : Dalhousie University

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Variable analysis

independent variables

Intestinal tissue preparation method (fixed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin)

dependent variables

Villus height
Villus width
Crypt depth
Total mucosa thickness (villus height + crypt depth)

control variables

Tissue sectioning thickness (0.5 μm)
Microscope magnification (4×)
Image processing and analysis system (Leica Application Suite, Version 3.4.0)

controls

No explicit positive or negative controls were mentioned in the provided information.

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