In the photosynthetic membrane, there is a particular aggregated state for the chlorophyll a (Chl a) molecules with a specific arrangement responsible for the high efficiency of energy conversion. Chl a monolayers, transferred onto solid substrates, are systems that potentially can mimic the packing of the in vivo system. The association of Chl a in the monolayer results in the formation of dimers with an average size of 3.00 ؎ 0.15 nm. Considering the organization of the dimers, we assume that P680 is a dimer with the (anti) parallel transition moments of the constituent. The Chl a macrocycles most likely are tilted to each other by 30؇ with respect to the membrane plane.Green plants use light energy, through photosynthesis, to reduce CO 2 and produce carbohydrates. The central unit where this activity takes place, the photosynthetic membrane, is described as a segment of lipid bilayer in which proteinpigment complexes are bound. Two complexes within the photosynthetic membrane are considered the key reaction centers, photosystem I (PS I) and PS II (1). Pigments, such as chlorophyll a (Chl a), located within PS I and PS II, play a major role in the capture of light energy and the subsequent charge transfer that results in CO 2 reduction (2). Chl a has a broad absorption spectrum, and aggregation through selfassembly typically leads to changes in its optical properties (3, 4). Red shifts are commonly observed in in vitro Chl a systems, such as thin films, monolayers, and colloidal dispersions, used as models for the in vivo system (5). One such system, based on the Langmuir-Blodgett (L-B) (6) technique for forming ordered thin films, allows the orientation of molecules in a molecular monolayer at an air͞water interface and the subsequent transfer onto solid substrates. The transferred L-B film is highly packed and well organized, and the use of this technique allows preparation of systems to potentially mimic the in vivo packing of Chl a within the photosynthetic membrane.Numerous attempts have been made to model the arrangement of Chl a to rationalize its high energy conversion efficiency. One model (7) proposed for the primary electron donor of PS I describes the formation of a Chl a dimer held together via water bridges. In this model, the oxygen of one H 2 O molecule is coordinated to the magnesium of a Chl a molecule, while its hydrogen bonds to the keto carbonyl group of a second Chl a molecule. A second H 2 O molecule completes the dimer in the same way. This results in a structure where the distance between the Mg centers is 0.89 nm, and the interplanar separation between the macrocycles is 0.36 nm.In an effort to obtain direct structural information on the aggregation of Chl a in thin films, we have examined monolayers of Chl a transferred onto freshly cleaved graphite (HOPG, ZYB grade, Advanced Ceramics, Lakewood, OH) by scanning tunneling (STM; Nanoscope II STM, Digital Instruments, Santa Barbara, CA) (8, 9) and atomic force (AFM) (10, 11) microscopies, i.e. scanning probe microscopy. Chl a (T...