We demonstrate the use of multiphoton-excited photochemistry to cross-link three-dimensional matrices directly from cytoplasmic proteins in a live cell (starfish oocyte). Fluorescence recovery after photobleaching measurements were used to determine diffusion coefficients inside intracellular cross-linked structures, and it was found that the diffusion was approximately 3 to 4 orders of magnitude slower than in free solution and 2-3 orders of magnitude slower than in cytoplasm and that the value can be tuned by controlling the laser exposure. Complex structures can be fabricated to construct channels and compartments that could be used to isolate cellular processes, and the method should thus be applicable to a broad range of problems in cell biology.
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