Dipteran chromosomes: a simple method for obtaining high quality chromosomal preparations

摘要

For cytogenetic studies it is necessary to develop an efficient method of chromosome preparation with well-preserved and dispersed chromosomes and little or no cell-wall debris. In this study, we describe a method for obtaining high quality chromosomal preparations of dipteran cells. This procedure combines drop method and air-dry technique and produces a cell suspension with separated and mixed cells. Suspended mitotic cells were then dropped onto a glass slide in order to distribute the cytoplasm and spread the chromosomes. We compare and evaluate different chromosome preparation methods to develop a more reliable procedure for the resolution of chromosome characteristics and chromosome bandings in blow fly mitotic chromosomes: The combined drop and air-dry technique described here is convenient for identification of sex chromosomes. Using this protocol, acceptable high-quality chromosomal spreads with flattened cells and no cell debris, and without damage and/or loss of chromosomes were obtained.