Counting Iron-absorbed Small Intestinal

Background:

To better study iron absorption in premature infants, the Kling lab is planning to quantify intestinal iron content in newborn rats fed different forms and different amounts of iron. In one group, the pups are dam fed. In the other group, the pups receive rat milk substitute (RMS) diet. The purpose of having these two groups is to study the differences in iron status depending on the source of the diet. Several measures of iorn status are employed; i.e. plasma iron, red cell iron, and tissue iron (Dubuque 2002). After experimental feeding, intestinal tissues will be harvested and prepared on microscope slides. These slides will be stained for iron and evaluated either microscopically (hand counting) or digitally to quantify the iron content in the small intestinal villi

Hand counting is the standard method used when analyzing tissue slides. However, the process is very time consuming, subjective, and error-prone. When hand counting, the number of villi stained positively for iron, i.e. blue, and the number of individual lining cells that contain blue stain within each villi will be counted. Some cells or regions of a villi are densly blue, where other cells or regions of the same villi contain a speck of blue, indicating very little iron. Currently, it is difficult to quantify this varying degree of iron staining. Hence, digital imaging methods are being investigated. Digital methods are available and being used in scientific literature in other research labs on campus and elsewhere (Ruifrok, 1997).

For more details, please read a report by Kelsey Kleven, titled: Analyzing for the Presence of Iron in Small Intestinal Villi.

Objectives

To develop an image processing program to automate the villi counting process and compare the result with the ImageJ-based prior result.

Reference Images