Antifungal Susceptibility Test via Single-Cell Morphology, Development, and Metabolism

Antifungal susceptibility test (AFST) is essential for
guiding
effective therapy, but it remains challenged by long turnaround times
and the limited generalizability of single-parameter phenotypic readouts
across diverse fungal species and antifungal classes. Here, we present
a multifeature AFST (MAFST) platform that integrates single-cell Raman
spectroscopy and brightfield imaging to enable rapid and pleiotropic
assessment of antifungal responses. By unifying complementary metabolic
and morphological features into a composite inhibition index (CII),
MAFST captures coordinated cellular responses to antifungal exposure
that cannot be reliably resolved by individual metrics alone. Benchmarking
against the broth microdilution demonstrates that CII-based MAFST
achieves high concordance across multiple Candida species and antifungal classes while substantially reducing the
time required for susceptibility determination. Thus, by capturing
single-cell morphological, developmental, and metabolic drug responses
in a fast, accurate, and widely applicable fashion, MAFST can contribute
to tackling fungal infections.