Efficient image cytometry of a conventional microscope slide means rapid acquisition and analysis of 20 gigapixels of image data (at 0.3-microm sampling). The voluminous data motivate increased acquisition speed to enable many biomedical applications. Continuous-motion time-delay-and-integrate (TDI) scanning has the potential to speed image acquisition while retaining sensitivity, but the challenge of implementing high-resolution autofocus operating simultaneously with acquisition has limited its adoption. We develop a dynamic autofocus system for this need using: 1. a "volume camera," consisting of nine fiber optic imaging conduits to charge-coupled device (CCD) sensors, that acquires images in parallel from different focal planes, 2. an array of mixed analog-digital processing circuits that measure the high spatial frequencies of the multiple image streams to create focus indices, and 3. a software system that reads and analyzes the focus data streams and calculates best focus for closed feedback loop control. Our system updates autofocus at 56 Hz (or once every 21 microm of stage travel) to collect sharply focused images sampled at 0.3x0.3 microm(2)/pixel at a stage speed of 2.3 mms. The system, tested by focusing in phase contrast and imaging long fluorescence strips, achieves high-performance closed-loop image-content-based autofocus in continuous scanning for the first time.
Diffuse reflectance near infrared hyperspectral imaging is an important analytical tool for a wide variety of industries, including agriculture 1,2 , consumer products, chemical 3 and pharmaceutical development and production 4,5 . Using this technique as a method for the standoff detection of explosive particles is presented and discussed. The detection of the particles is based on the diffuse reflectance of light from the particle in the near infrared wavelength range where CH, NH, OH vibrational overtones and combination bands are prominent.The imaging system is a NIR focal plane array camera with a tunable OPO/ laser system as the illumination source. The OPO is programmed to scan over a wide spectral range in the NIR and the camera is synchronized to record the light reflected from the target for each wavelength. The spectral resolution of this system is significantly higher than that of hyperspectral systems that incorporate filters or dispersive elements. The data acquisition is very fast and the entire hyperspectral cube can be collected in seconds. A comparison of data collected with the OPO system to data obtained with a broadband light source with LCTF filters is presented.
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