Future Research

Future Research (as visiting scientist)

Specific Aims

• Place cells on a microscope slide to reproduce the chromatin circles and structures as shown on the blog.

• Preserve the structures so they can be further examined.

1) Freeze the slides on dry ice to remove the coverslip and preserve the structures.

2) Heat fix both the coverslip and slide to preserve the structures.

• Determine the chemical nature of their composition.

1) Perform in situ immunostaining on the structures to determine the following:

a) The general nature of their composition: protein, phospholipid, and nucleic acids

b) Specific types of biomolecules associated with origins of replication like topoisomerase II

2) Changes in the composition of the structures as they pass through the cell cycle or engage in apoptosis and necrosis.

• Study the physical structure of the circles using transmission electron microscopy (TEM).

1) Place cells onto an electron microscope grid under a microscope slide and coverslip and allow the  circles to develop.

2) Freeze the slides on dry ice to remove the coverslip and grid and preserve the structures.

3) Heat fix the grid to preserve the structures.

4) Stain and/or shadow the grid for TEM.

5) Examine the sample by TEM to check for connections between various structures that could be DNA or other linkages.

•  Use in situ DNA probes for the following studies:

1) Determine the structure and location of specific genes within these circles.

2) Determine the effect of the cell cycle, apoptosis, and necrosis on the structure and location of specific genes within these circles.

3) Determine the developmental fate of specific genes within these circles during cellular differentiation.

4) Locate and determine the developmental fates of origins of replication, promoters, and enhancers within these circles.