RESUMEN
Localization microscopy relies on computationally efficient Gaussian approximations of the point spread function for the calculation of fluorophore positions. Theoretical predictions show that under specific experimental conditions, localization accuracy is significantly improved when the localization is performed using a more realistic model. Here, we show how this can be achieved by considering three-dimensional (3-D) point spread function models for the wide field microscope. We introduce a least-squares point spread function fitting framework that utilizes the Gibson and Lanni model and propose a computationally efficient way for evaluating its derivative functions. We demonstrate the usefulness of the proposed approach with algorithms for particle localization and defocus estimation, both implemented as plugins for ImageJ.
Asunto(s)
Algoritmos , Imagenología Tridimensional/métodos , Microscopía Fluorescente/métodos , Biología Computacional/métodos , Colorantes Fluorescentes/química , Análisis de los Mínimos Cuadrados , Fotones , Distribución de Poisson , Refractometría , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
268 patients with suspected lung cancer were examined over the period 1 January 1972 to 31 May 1974. On the basis of the 735 investigations performed, a 6-8 h workup program has been established including physical and functional examination, standard X-rays and hilar tomography, endoscopy and biopsy for cytologic or histologic study. Several biopsy technics are described and their indications tabulated according to their efficiency in relation to the radiologic aspect of the lesion. The number of cases and the yield for each technic are summarized in Table 2.
