Human Lymphocyte Metaphase Chromosome Preparation for Radiation-Induced Chromosome Aberration Analysis.

Abstract

Radiation-induced chromosomal aberration analysis for metaphase chromosomes is well established and the golden standard for human biodosimetry. This method can estimate doses of human radiation exposure after nuclear accident and unwanted radiation exposure from their lymphocytes. The natural background frequency of dicentric chromosome for human lymphocytes is less than 1% and any increase in dicentric and centric ring chromosomes may be highly associated with radiation exposure. With the appropriate number of metaphase cells, one can detect the exposure of more than 0.1Gy by observing dicentric and centric ring chromosomes. Dicentric chromosome analysis is relying on morphological changes and may be difficult for researchers without appropriate training. This method is time consuming and labor intensive, but still currently the most reliable technique and analysis needs only light microscopes with high magnification objectives and trained personnel. Recent research enables us to visualize dicentric chromosomes clearly with fluorescent markers for easy detection of dicentric and centric ring chromosomes. This chapter will introduce classical dicentric analysis of human lymphocyte cells with Giemsa staining.