Accurate measurement of the moisture content of paper is essential in papermaking and is also important in some paper-based microfluidic devices. Traditional measurement techniques provide very limited spatiotemporal resolution and working range. This article presents a novel method for moisture content measurement whose operating principle is the strong correlation between the optical transparency of paper and its moisture content. Spectrographic and microscopic measurement techniques were employed to characterize the relation of moisture content and relative transparency of four types of paper: hardwood chemi-thermomechanical pulp paper, Northern bleached softwood kraft paper, unbleached softwood kraft paper, and General Electric(®) Whatman™ grade 1 chromatography paper. It was found that for all paper types, the paper transparency increased monotonically with the moisture content (as the ratio of the mass-of-water to the mass-of-dry-paper increased from 0% to 120%). This significant increase in relative transparency occurred due to the refractive index matching role of water in wet paper. It is further shown that mechanical loading of the paper has little impact on the relative transparency, for loadings that would be typical on a paper machine. The results of two transient water absorption experiments are presented that show the utility and accuracy of the technique.