The laser calorimetry system has been used previously to assess laser heating of a series of GNS, GNR, and gold nanoshell solutions9,35,36. Freshly prepared GNS suspensions are loaded into a four-side plastic optical cuvette and heated with a 532 nm CW laser (190 mW). To minimize measurement errors, four temperature thermocouples and a magnetic stirrer were used. The GNS suspension was photoheated to steady state (< 0.1 °C temperature change in 1 min) and a maximum temperature rise was measured (∆T, °C). A resistive heater with known power was placed in the cuvette system and used to calibrate the specific absorption rate (SAR) assuming the laser GNS heat source of the same power produced the same temperature rise. Resistive heater calibration was performed multiple times over a wide power range (25–250 mW). Calibrated in a previous study9, there was a linear correlation between ∆T and the experimental photothermal heat generation (Pexp, mW) with a fitting coefficient (Eq. (10)):

Cuvette calorimetry measurements were conducted three times for each GNS aggregate suspension under the same conditions.

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