Membrane fluidity can be affected by a number of factors. One way to increase membrane fluidity is to heat up the membrane. Lipids acquire thermal energy when they are heated up; energetic lipids move around more, arranging and rearranging randomly, making the membrane more fluid. At low temperatures, the lipids are laterally ordered and organized in the membrane, and the lipid chains are mostly in the all-trans configuration and pack well together.
The composition of a membrane can also affect its fluidity. The membrane phospholipids incorporate fatty acids of varying length and saturation. Lipids with shorter chains are less stiff and less viscous because they are more susceptible to changes in kinetic energy due to their smaller molecular size and they have less surface area to undergo stabilizing van der Waals interactions with neighboring hydrophobic chains. Lipid chains with carbon-carbon double bonds (unsaturated) are more fluid than lipids that are saturated with hydrogens and thus have only single bonds. On the molecular level, unsaturated double bonds make it harder for the lipids to pack together by putting kinks into the otherwise straightened hydrocarbon chain. Membranes made with such lipids have lower melting points: less thermal energy is required to achieve the same level of fluidity as membranes made with lipids with saturated chains. Incorporation of particular lipids, such as sphingomyelin, into synthetic lipid membranes is known to stiffen a membrane. Such membranes can be described as "a glass state, i.e., rigid but without crystalline order".
Cholesterol acts as a bidirectional regulator of membrane fluidity because at high temperatures, it stabilizes the membrane and raises its melting point, whereas at low temperatures it intercalates between the phospholipids and prevents them from clustering together and stiffening. Some drugs, e.g. Losartan, are also known to alter membrane viscosity. Another way to change membrane fluidity is to change the pressure. In the laboratory, supported lipid bilayers and monolayers can be made artificially. In such cases, one can still speak of membrane fluidity. These membranes are supported by a flat surface, e.g. the bottom of a box. The fluidity of these membranes can be controlled by the lateral pressure applied, e.g. by the side walls of a box.
Discrete lipid domains with differing composition, and thus membrane fluidity, can coexist in model lipid membranes; this can be observed using fluorescence microscopy. The biological analogue, 'lipid raft', is hypothesized to exist in cell membranes and perform biological functions. Also, a narrow annular lipid shell of membrane lipids in contact with integral membrane proteins have low fluidity compared to bulk lipids in biological membranes, as these lipid molecules stay stuck to surface of the protein macromolecules.
Membrane fluidity can be measured with electron spin resonance (ESR), fluorescence, or deuterium nuclear magnetic resonance spectroscopy (NMR). ESR measurements involve observing spin probe behaviour in the membrane. Fluorescence experiments involve observing fluorescent probes incorporated into the membrane. Solid state deuterium nuclear magnetic resonance spectroscopy involves observing deuterated lipids. The techniques are complementary in that they operate on different timescales.