Comparison of two-dimensional fast Raman imaging versus point-by-point acquisition mode for human bone...

Recent technical developments gave rise to a new technology for two-dimensional fast Raman imaging: the DuoScan averaging mode (DS-Avg). This technology allows the acquisition of a Raman spectrum over a rastered macro spot. The aim of this study was to evaluate the interest of the DS-Avg applied on trabecular human bone. The evaluation was based on the comparison of the DS-Avg versus the point-by-point mapping mode in real usage conditions. The signal-to-noise ratio, the spectral difference, and the physicochemical parameters were estimated for comparison of the efficiency of both modes. Principal component analysis was performed to explore the capacity of both modes to detect compositional variations. Results showed that the DS-Avg spectrum was equivalent to the average spectrum of individual spectra acquired with the point-by-point mode for the same sample area. The physicochemical parameters can be also determined from DS-Avg acquisition. The DS-Avg combined with an objective ×50 allows a drastic decrease of the acquisition time, but the information about the micrometric composition is lost. The combination of the DS-Avg with an objective ×100 is a good compromise between acquisition time and resolution. The DS-Avg is a useful technology for imaging mineral and organic phases of bones and for assessing their spatial distribution on large samples. The point-by-point imaging mode is more appropriate to assess the heterogeneous composition of bone within the micrometer scale. For the first time, this study compares the DuoScan averaging mode to the point-by-point imaging mode on a trabecular human bone.