Measurement technique, point counting

Coincident with this new technique for procurement of human bone biopsies was the development of quantitive methods of bone analysis.12 These methods include histochemical analysis of both decalcified and unde-calcified42 48 bone sections, microradiography,44 tetracycline labeling45 and autoradiography.42 The latter two techniques require administration of a tetracycline antibiotic or isotopic tracer prior to procurement of the biopsy. Undecalcified thin sections, prepared with the use of a Jung microtome after the bone core is fixed, dehydrated and embedded in methacrylate,45 are analyzed by intersect and point count methods46 47 which permit three-dimensional assessment.48 49 Tetracycline antibiotics deposit in vivo in sites of bone formation constituting markers which can be studied in undecalcified sections by fluorescence microscopy.45 47 This represents the safest and best tissue time marker for microscopic measurement of bone formation dynamics. 

Iodine-129 is a weak emitter with end-point energy of 152 keV and decays to Xe that emits 40 keV y and X-rays. Due to its long half-Kfe, it is difficult to measure by decay counting. The concentration of 1 in natural samples, for example, seawater, is extremely low ( 10 atoms/g) relative to 1 and can only be measured by AMS, the atom counting technique, as INAA and ICP-MS are limited by lower sensitivity. 

Because X-ray counting rates are relatively low, it typically requires 100 seconds or more to accumulate adequate counting statistics for a quantitative analysis. As a result, the usual strategy in applying electron probe microanalysis is to make quantitative measurements at a limited collection of points. Specific analysis locations are selected with the aid of a rapid imaging technique, such as an SEM image prepared with backscattered electrons, which are sensitive to compositional variations, or with the associated optical microscope. 

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