High-speed hyperspectral Raman imaging for label-free compositional microanalysis

Biomed Opt Express. 2013 Oct 8;4(11):2376-82. doi: 10.1364/BOE.4.002376. eCollection 2013.

Abstract

We present high-speed hyperspectral Raman imaging with integrated active-illumination for label-free compositional microanalysis. We show that high-quality Raman spectra can be acquired from as many as ~1,000 spots/sec semi-randomly distributed among a ~100x100 μm(2) area without mechanical scanning. We demonstrate rapid data acquisition from three types of samples: 1) uniform, strong Raman scatterers, e.g., silicon substrates; 2) non-uniform, medium-strength Raman scatterers, e.g., polymer microparticles; and, 3) non-uniform, relatively weak Raman scatterers, e.g., bacterial spores. We compare the system performance to that of point-scan with an electron-multiplied CCD camera, as implemented in some commercial systems. The results suggest that our system not only provides significant imaging speed advantage for various types of samples, but also permits substantially longer integration time per spot, leading to superior signal-to-noise ratio data. Our system enables the rapid collection of high quality Raman spectra for reliable and robust compositional microanalysis that are potentially transformative in applications such as semiconductor material and device, polymer blend and biomedicine.

Keywords: (170.0110) Imaging systems; (170.1530) Cell analysis; (180.5655) Raman microscopy.