Precise and millidegree stable temperature control for fluorescence imaging: Application to phase transitions in lipid membranes

We present the design of a custom temperature-controlled chamber suitable for water or oil immersion fluorescence microscopy and its application to phase behavior in lipid bilayer vesicles. The apparatus is self-contained and portable, suitable for multiuser microscopy facilities. It offers a higher temperature resolution and stability than any comparable commercial apparatus, on the order of millidegrees. We demonstrate the utility of the system in the study of miscibility transitions in model membranes. The temperature-dependent phase behavior of model membrane systems that display liquid-ordered ( L o ) phase coexistence with the liquid-disordered ( L d ) phase is relevant to understanding the existence of heterogeneities in biological cell plasma membranes, ubiquitously termed "lipid rafts."