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The non-invasive optical technique of near-infrared spectroscopy (NIRS) was used to measure the depths of light penetration at four wavelengths in the NIR region. Near-infrared absorbance measurements were carried out on ten preterm infants. The apparent absorbance data (in optical density units) collected at 775, 805, 845, and 904 nm were measured at different positions on the head. Linear relationships that satisfy the Lambert law were obtained when the apparent absorbance at a given wavelength was plotted against the inter-optrode distance. From the slopes of the resulting straight lines, the depth of penetration of the NIR light was calculated and found to be independent of the position of the probes on the head. Calculated average values of the depth of penetration of near-infrared light in the whole neonatal head ranged between 6.3 and 8.5 mm.