The 'reference trap' revisited: examples of the dangers in using ratios to describe fetoplacental angiogenesis and trophoblast turnover.

Examples of the use of relative or ratio data are to be found throughout biomedical sciences and include such variables as stereological component densities per cell volume (morphology), transport rates per tissue volume or membrane surface (physiology), gold labelling frequencies (immunocytochemistry) and amounts of DNA relative to protein (biochemistry). This report emphasizes the potential pifalls associated with drawing biological conclusions and interpretations from relative data when there has been no attempt to monitor the absolute size of the reference space. This is know as the 'reference trap'. The dangers to interpretation inherent in confining results to such data are illustrated using the sorts of data found in current placental studies on fetoplacental angiogenesis and villous trophoblast turnover but are pertinent to many other areas. Where it is possible to do so, these dangers can be avoided by the simple expedient of estimating the size of the pertinent reference space and using this to calculate absolute values. Sometimes (e.g. when relying on biopsy samples), the size of the reference cannot be determined. In such cases, ratio data must be interpreted with due caution.