Raman Molecular Imaging: A Novel Technique for Histopathological Evaluation of Tissue Specimens
JS Maier, AJ Drauch, SD Stewart. ChemImage Corporation, Pittsburgh, PA
Background: Raman Molecular Imaging (RMI) is a digital, non-destructive, reagentless tissue imaging approach based on Raman spectroscopy. RMI provides an image of a sample wherein the contrast derives from Raman scattering of light off of the constituent molecules of the tissue.
Design: Thin sections from formalin fixed, paraffin-embedded samples were obtained in standard fashion and applied to aluminum coated microsope slides. Digital Raman images were acquired using a FALCON™ Raman imaging system. After Raman imaging, samples were stained to facilitate histological correlation between traditional H&E and Raman imaging modalities. Digital images were created by process of constructing a reference library of spectra for selected histological types, and subsequent segmenting using spectral mixture resolution. Analysis included subjective comparison of Raman images to imagery of the stained sample.
Results: Images show good qualitative correlation between the reagentless RMI and the images of tissue stained in traditional fashion in terms of histological features including prostate epithelium and stroma, renal glomerular cells and tubular cells, and red blood cells.
Figure 1 shows an H&E stained field of view from a prostate.
Figure 2 shows the image resulting from spectral mixture resolution applied to RMI of the same field of view of prostate prior to staining.
Conclusions: Raman Molecular Imaging (RMI) is a novel technique for digital image based analysis of unstained thin sections of tissue. RMI carries information about the molecular environment at the cellular and subcellular level in tissues as manifested by accurate histological sectioning based on comparison to existing, stain-based approaches. The molecular information at the basis of the contrast in RMI may have correlation with pathophysiology that is of clinical interest. Further study of RMI in application to clinical questions of interest is warranted.
Wednesday, March 24, 2010 9:30 AM
Poster Session V # 222, Wednesday Morning