Authors
Albert L, Basak S, Koerner H, Oleksiievets N, Mougios N, Cotroneo ER, Frei MS, Enderlein J, Broichhagen J, Simeth NA, Tsukanov R, Opazo F
Journal
BioRxiv
Citation
bioRxiv 2026.03.02.709025.
Abstract
Multiplexed fluorescence imaging is limited by spectral overlap, whereas fluorescence lifetime provides an orthogonal encoding dimension. We replace genetically fused HaloTags with recombinant nanobody-HaloTag constructs applied as immunofluorescence reagents and determine a lifetime palette. Integrating lifetime and spectral encoding enables up to eight targets in a single acquisition and supports multiplexed imaging in cells and tissue using a simple OneStep labeling strategy compatible with fluorescent proteins, STED nanoscopy, and any standard laboratory antibody.
