Mid-infrared Chemical Nano-imaging for Intra-cellular Drug Localisation

TL;DR

This paper introduces Mid-infrared Chemical Nano-imaging (MICHNI), a novel technique enabling label-free, high-resolution chemical mapping of drugs within cells, overcoming limitations of existing microscopy methods.

Contribution

The authors adapt probe-based infrared techniques for routine biological samples, achieving ~10 nm resolution for cellular chemical imaging, including drug localization.

Findings

Successfully mapped Bortezomib within a single human cell.

Demonstrated compatibility with standard biological specimen preparation.

Potential to complement electron microscopy in bio-sciences.

Abstract

In the past two decades a range of fluorescence cell microscopy techniques have been developed which can achieve ~10 nm spatial resolution, i.e. substantially beating the usual limits set by optical diffraction. However, these methods rely on specialised labelling. This limits the applicability, risks perturbing the biology, and it also makes them so-called "discovery techniques" that can only be used when there is prior knowledge about the biological problem. The alternative, electron microscopy (EM), requires complex and time-consuming sample preparation, that risks compromising the sample's integrity. Samples have to withstand vacuum, and staining with heavy metals to make them conductive, and give usable electron-contrast. None of these techniques can directly map out drug distributions at a sub-cellular level. Recently infrared light-based scanning probe techniques have demonstrated a capability for ~1 nm spatial resolution. However, they need samples that are flat, dry and dimensionally stable and they only probe down to a depth commensurate with the spatial resolution, so they yield essentially surface chemical information. Thus far they have been applied only to artificially produced test samples, e.g. gold particles, or isolated proteins on silicon. Here we show how these probe-based techniques can be adapted for use with routinely prepared general biological specimens. This allows for "Mid-infrared Chemical Nano-imaging" (MICHNI) that delivers chemical analysis at a ~10 nm spatial resolution, suitable for studying cellular ultrastructure. We demonstrate its utility by performing label-free mapping of the anti-cancer drug Bortezomib (BTZ) within a single human myeloma cell. We believe that this MICHNI technique has the potential to become a widely applicable adjunct to EM across the bio-sciences.