This paper describes analytical methods for the determination of calcium, phosphate and fluoride in successive layers of enamel removed from rat and human teeth by acid etching. These methods include: (i) simple, sensitive spectrophotometric methods for the determination of calcium and phosphate; (ii) an easily constructed miniature fluoride electrode, and (iii) an improved electrode adapter that permits one fluoride electrode to measure numerous microsamples very rapidly. The high sensitivity and simplicity of the analytical techniques described allows many types of microsamples of interest in dental research to be analyzed with an accuracy and speed heretofore attainable only with much larger samples.