Background correction in forensic photography. I. Photography of blood under conditions of non-uniform illumination or variable substrate color--theoretical aspects and proof of concept

The combination of photographs taken at two or three wavelengths at and bracketing an absorbance peak indicative of a particular compound can lead to an image with enhanced visualization of the compound. This procedure works best for compounds with absorbance bands that are narrow compared with "average" chromophores. If necessary, the photographs can be taken with different exposure times to ensure that sufficient light from the substrate is detected at all three wavelengths. The combination of images is readily performed if the images are obtained with a digital camera and are then processed using an image processing program. Best results are obtained if linear images at the peak maximum, at a slightly shorter wavelength, and at a slightly longer wavelength are used. However, acceptable results can also be obtained under many conditions if non-linear photographs are used or if only two wavelengths (one of which is at the peak maximum) are combined. These latter conditions are more achievable by many "mid-range" digital cameras. Wavelength selection can either be by controlling the illumination (e.g., by using an alternate light source) or by use of narrow bandpass filters. The technique is illustrated using blood as the target analyte, using bands of light centered at 395, 415, and 435 nm. The extension of the method to detection of blood by fluorescence quenching is also described.