Isolation of epidermal lamellar bodies has presented a challenge because pressures required to homogenize keratinocytes can destroy these organelles and because the lamellar body readily releases its contents during prolonged isolation procedures. In an attempt to isolate lamellar bodies, sheets of intact stratum corneum and stratum granulosum were obtained from neonatal mice with highly purified staphylococcal epidermolytic toxin, disrupted, and passed through a series of filters. The final filtrate was rich in intact lamellar bodies and contained variable amounts of ribosomes and other vesicular structures. Availability of a highly purified lamellar body preparation from postnatal epidermis should help to clarify the role of this organelle in epidermal function. The technique of selective, sequential filtration represents a new approach to cell fractionation that may have wide applications in cell biology and biochemistry.