Autonomous technologies are needed which are capable of sensing real time changes in biophysical transport across cell membranes/organelles. These technologies must not only be highly sensitive/selective, but must also be minimally invasive/intrusive, causing no significant physical/chemical effects on cell behavior. Challenges with mainstream technologies (e.g., assays, fluorescent dyes, microsensors) include signal noise/drift, low temporal resolution, requirement of large sample sizes, cytoxicity, organelle sequestration, and intracellular buffering. Recent advancements in fiber optics have greatly enhanced the performance of microsensors (e.g., increased sensitivity/selectivity, response time), but used in concentration mode near cells/tissues these sensors suffer from poor signal to noise ratio. Work over the last few decades has advanced microsensor utility through sensing modalities that extend and enhance the data recorded by sensors. This technique, known as self-referencing, converts static micro/nanosensors with otherwise low signal-to-noise ratios into dynamic flux sensors capable of filtering out signals not associated with active transport by acquisition and amplification of differential signals. Here, we demonstrate the use of a self-referencing referencing frequency domain fiber optic microsensor containing a quenched dye (platinum tetrakis-pentafluorophenyl porphyrin) for quantifying cell/tissue flux in biomedical, agricultural, and environmental applications.
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