An analysis protocol must identify the general requirements for each specimen type. The protocol should identify normal parameters and normal variants and should distinguish between true abnormality and artifact. The number of cells from which chromosomes are to be counted, sex chromosomes examined, and bands analyzed in detail must be clearly stated, including whether each type of examination is to occur at the microscope, on an image, or via a karyotype. A protocol should set standards for the selection of suitable metaphase spreads, as well as the number of cultures (and colonies, when applicable) from which cells should be examined. When an abnormality is detected, the appropriate steps to take should be specified. Other items, such as an appropriate banding resolution level, maximum allowable number of overlapping chromosomes, random chromosome loss, and dealing with metaphases in close proximity, might also be included.
A protocol should identify the procedures used to document each metaphase, as well as the data to be recorded on a microscope analysis worksheet, requirements for imaging, the number of cells to be karyotyped, the number of individuals who should take part in performing the analysis, and the individual who should verify the results.
Finally, a protocol should establish the policies for the storage of microscope slides, both during analysis and once analysis has been completed.
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