Strike a Pose: Imaging and Cell Counting

The next step in the process of determining the effects of treatment on neural development is to visually count the cells in the developing nervous system. To do this, DAPI stained sections are imaged. From those images, a program counts the cells in the developing nervous system.

To refresh your memory, DAPI is a cell stain that binds to the minor groove of DNA. It is a photosensitive and fluorescent stain; therefore when the slides are stained with this solution, a minimal amount of light is permitted in the room.  Because the stain is purely fluorescent, there is no visible change to the sections.

In order to image the cells stained by DAPI, we use a scope with fluorescent imaging capabilities. Ultraviolet light excites the DAPI in the cells, so a scope with a UV filter and long exposure is used to visualize the DAPI in each section. From these images, ROI counts are made to determine the number of cells in each section. This program allows the user to set parameters so that only the cells we want to count are counted.

Counting cells is an important step my project. Because the hydroxyurea treatment should suppress cell division, the embryos that have undergone treatment should have a lower number of neural cells than their untreated, sibling controls. If the cell number in the treatment group is greater than or equal to the control, the hypothesis would be disproved. Results such as that would change the direction of my project, and possibly impact previous studies claiming the inhibitive effects of hydroxyurea treatment.

All in all, this is a critical step in the progression of this project. Once all of the slides are imaged and counted, a major piece of the puzzle will be complete.