Unbiased estimation of particle density in the tandem scanning reflected light microscope

The tandem scanning reflected light microscope has the property of being able to obtain information from 'inside' solid objects by taking a thin optical section at the focal plane of the objective lens. This plane can be focused up and down through the specimen. We describe an unbiased 3-D counting rule for the TSRLM, which is applied to the estimation of osteocyte lacunar density in whole bone. This is shown to be an extremely efficient way of making such an estimate. Further possibilities for the application of the microscope in the field of stereology are discussed.