Microenvironmental Fingerprint of Maturing Tau Condensates from Spectral and Lifetime Phasor Analysis of a Solvatochromic Dye

ACDAN phasor analysis shows Tau condensate maturation (1, 4, 48 h) is linked to a progressive decrease in spectral phasor angle {theta} and increase in phase-derived lifetime {tau}P.

• Why it matters: Spectral and lifetime readouts can respond differently to the same local environment, so separating these sensitivities is needed to track how Tau assembly microenvironments evolve over time.
• What they did: The authors built a multimodal phasor framework for two-photon spectral and fluorescence lifetime imaging, validated it with ACDAN in water, glycerol, and ethanol, then applied it to Tau condensates across 1, 4, and 48 h.
• The result: Differences among Tau condensates were apparent by 4 h, with pronounced spatial heterogeneity at 48 h, and Thioflavin T mainly reported via fluorescence lifetime while ACDAN captured changes in both coordinates.

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