RESUMEN

The purpose of this study is to investigate how to scale pixel intensity acquired from one exposure time to another. This is required when comparing grayscale images acquired at different exposure times and other image processing such as autofluorescence removal. Pixel intensity is linear to exposure time as long as images are acquired at the linear range of a camera, but importantly there exists an intercept, which is set by the camera. We termed this intercept as dark pixel intensity, as it is the pixel intensity under conditions of no light and zero exposure time. Dark pixel intensity is determined by camera's readout noise (electron/pixel), gain, and DC offset. Knowing dark pixel intensity, image acquired from one exposure time can be linearly scaled to an image at a different exposure time. Dark pixel intensity can be directly measured by obtaining an image at no light and zero (or minimum) exposure time. It can be also indirectly calculated by capturing images at a series of exposure times. Finally, the prestained and poststained images were acquired at their optimal exposures and autofluorescence was completely removed by normalizing images with the exposure time ratio and dark pixel intensity followed by subtraction.

Asunto(s)

Microscopía Fluorescente/métodos , Microscopía Fluorescente/normas , Patología/métodos , Patología/normas , Neoplasias de la Mama/patología , Humanos , Masculino , Piel/patología

RESUMEN

Laser ultrasound is now integrated into the manufacturing process of some of the most modern aircraft for the inspection of composite parts. Unfortunately, for some material and process combinations, laser-ultrasound suffers from a lack of sensitivity. In laser-ultrasound generation, optical penetration depth plays a very important role. It was shown that changing the generation wavelength from the 10.6 microm of the CO2 laser to the 3-4 microm range can significantly improve generation efficiency. In this paper, ultrasonic displacements are compared to measurements of optical penetration depth in different polymer-matrix composites. Ultrasonic waves were generated using an optical parametric oscillator operating in the 3.0-3.5 microm band and optical penetration depth spectra were evaluated using quantitative photoacoustic spectroscopy. The relative amplitudes of the generated ultrasonic waves track closely the optical penetration depth spectra. These results experimentally demonstrate the importance of optical penetration in the laser-ultrasound generation process.