In biology, membrane fluidity refers to the viscosity of the lipid bilayer of a cell membrane or a synthetic lipid membrane. Lipid packing can influence the fluidity of the membrane. Viscosity of the membrane can affect the rotation and diffusion of proteins and other bio-molecules within the membrane, thereby affecting the functions of these things.

Membrane fluidity is affected by fatty acids. More specifically, whether the fatty acids are saturated or unsaturated has an effect on membrane fluidity. Saturated fatty acids have no double bonds in the hydrocarbon chain, and the maximum amount of hydrogen. The absence of double bonds decreases fluidity. Unsaturated fatty acids have at least one double bond, creating a "kink" in the chain. The double bond increases fluidity. While the addition of one double bond raises the melting temperature, research conducted by Xiaoguang Yang et. al. supports that four or more double bonds has a direct correlation to membrane fluidity. Membrane fluidity is also affected by cholesterol. Cholesterol can make the cell membrane fluid as well as rigid.

Membrane fluidity can be affected by a number of factors. The main factors affecting membrane fluidity are environmental (ie. temperature), and compositionally. 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 melting temperature ${\displaystyle T_{m}}$ of a membrane is defined as the temperature across which the membrane transitions from a crystal-like to a fluid-like organization, or vice versa. This phase transition is not an actual state transition, but the two levels of organizations are very similar to a solid and liquid state.

The composition of a membrane can also affect its fluidity. The membrane phospholipids incorporate fatty acyl chains 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 London forces with neighboring hydrophobic chains. Molecules with carbon-carbon double bonds (unsaturated) are more rigid than those that are saturated with hydrogens, as double bonds cannot freely turn. As a result, the presence of fatty acyl chains with unsaturated double bonds makes it harder for the lipids to pack together by putting kinks into the otherwise straightened hydrocarbon chain. While unsaturated lipids may have more rigid individual bonds, membranes made with such lipids are more fluid because the individual lipids cannot pack as tightly as saturated lipids and thus have lower melting points: less thermal energy is required to achieve the same level of fluidity as membranes made with lipids with saturated hydrocarbon 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, fluorescence, atomic force microscopy-based force spectroscopy, or deuterium nuclear magnetic resonance spectroscopy. Electron spin resonance measurements involve observing spin probe behaviour in the membrane. Fluorescence experiments involve observing fluorescent probes incorporated into the membrane. Atomic force microscopy experiments can measure fluidity on synthetic or isolated patches of native membranes. Solid state deuterium nuclear magnetic resonance spectroscopy involves observing deuterated lipids. The techniques are complementary in that they operate on different timescales.