Matrix material was extracted from biofilms of Candida albicans and Candida tropicalis and analysed chemically. Both preparations contained carbohydrate, protein, hexosamine, phosphorus and uronic acid. However, the major component in C. albicans matrix was glucose (32 %), whereas in C. tropicalis matrix it was hexosamine (27 %). Biofilms of C. albicans were more easily detached from plastic surfaces by treatment with the enzyme lyticase (β-1,3-glucanase) than were those of C. tropicalis. Biofilms of C. albicans were also partially detached by treatment with proteinase K, chitinase, DNase I, or β-N-acetylglucosaminidase, whereas C. tropicalis biofilms were only affected by lipase type VII or chitinase. To investigate a possible role for the matrix in biofilm resistance to antifungal agents, biofilms of C. albicans were grown under conditions of continuous flow in a modified Robbins device (MRD). These biofilms produced more matrix material than those grown statically, and were significantly more resistant to amphotericin B. Biofilms of C. tropicalis synthesized large amounts of matrix material even when grown statically, and such biofilms were completely resistant to both amphotericin B and fluconazole. Mixed-species biofilms of C. albicans and a slime-producing strain of Staphylococcus epidermidis (RP62A), when grown statically or in the MRD, were also completely resistant to amphotericin B and fluconazole. Mixed-species biofilms of C. albicans and a slime-negative mutant of S. epidermidis (M7), on the other hand, were completely drug resistant only when grown under flow conditions. These results demonstrate that the matrix can make a significant contribution to drug resistance in Candida biofilms, especially under conditions similar to those found in catheter infections in vivo, and that the composition of the matrix material is an important determinant in resistance.