Linear Variable Filter‐Based Wide‐Field Hyperspectral Camera for Multiplex Immunoplasmonics Imaging

ABSTRACT

Hyperspectral imaging combined with plasmonic nanoparticles offers powerful opportunities for nanoscale biological diagnostics, yet conventional push‐broom systems are limited by slow acquisition and reduced spatial resolution. We report a wide‐field hyperspectral imaging platform integrating a custom linear variable tunable filter (LVTF) with reflected light microscopy (RLM) for multiplexed plasmonic nanoparticle (NP) imaging. While LVTF‐based systems have been used in industrial contexts, their adaptation to RLM microscopy enables high‐resolution, high‐contrast spectral imaging of NPs on biological samples. The system acquires data across 380–720 nm with 7–20 nm spectral resolution while preserving spatial detail. We demonstrate imaging of NPs bound to cancer cell membranes and classification of four distinct NP types. A trained support vector machine achieves an overall F ‐score of ~97%, significantly outperforming RGB‐based approaches. Compared to push‐broom systems, this platform provides faster acquisition, improved spatial resolution, and a compact, cost‐effective solution for multiplex NP detection in bioimaging and immunoplasmonic applications.