Cast silk membranes exhibit useful properties. However, there is limited control over the molecular architecture in these structures. The Langmuir-Blodgett technique can enhance the control of the membrane structure and allow improved control over membrane properties. We have formed natural silk monolayers using the Langmuir technique. Silk fibroin, regenerated from Bombvx mori cocoons, formed stable monolayers evident from pressure/area isotherms. Multilayers of the silk fibroin monolayers were deposited on a number of substrates and characterized. Transmission Electron Microscopy (TEM) and ellipsometry data provide basic information about the physical characteristics of the monolayer. Preliminary analysis of electron diffraction data of the monolayer indicate polycrystalline structure, consistent with the known structure of silk. Infrared spectrometric analysis of the monolayer using Attenuated Total Reflectance (ATR) gave wavenumbers for Amide I, II, III and V bands which compare with the silk II conformation reported for cast silk membranes.