A label-free approach by infrared spectroscopic imaging for interrogating the biochemistry of diabetic nephropathy progression

Abstract

Routine histology, the current gold standard, involves staining for specific biomolecules. However, untapped biochemical information in tissue can be gathered using biochemical imaging. Infrared spectroscopy is an emerging modality that allows label-free chemical imaging to derive biochemical information (such as protein, lipids, DNA, collagen) from tissues. Here we employed this technology in order to better predict the development of diabetic nephropathy. Using human primary kidney biopsies or nephrectomies, we obtained tissue from four histologically normal kidneys, four histologically normal kidneys from diabetic subjects and five kidneys with evidence of diabetic nephropathy. A biochemical signature of diabetic nephropathy was derived that enabled prediction of nephropathy based on the ratio of only two spectral frequencies. Nonetheless, using the entire spectrum of biochemical information, we were able to detect renal disease with near perfect accuracy. Additionally, study of sequential protocol biopsies from three transplanted kidneys showed biochemical changes even prior to clinical manifestation of diabetic nephropathy. Thus, infrared imaging can identify critical biochemical alterations that precede morphological changes, potentially allowing for earlier intervention.

Multi-radial LBP Features as a Tool for Rapid Glomerular Detection and Assessment in Whole Slide Histopathology Images

“…Most reported approaches have shown relatively low precision, have required access to extremely large datasets [20,21], or have made use of biochemical markers. Regarding the latter, in addition to the immunohistochemical and MRI methods earlier alluded to, one particularly novel example involves the use of Fourier-transform IR spectroscopy to assess the progression of diabetic nephropathy in pre-selected glomeruli [30]; however, the sample size was relatively small (68 glomeruli), and the method is likely not conveniently applied to glomerular detection as such.…”

Label-free molecular imaging of the kidney

“…179 developed an IR-MS imaging tool for diagnosing DN at a very early stage. The authors analyzed kidney biopsies from normal nondiabetic patients, diabetic patients without DN, and DN patients focusing on 3 structures of the extracellular matrix (i.e., mesangium, glomerular basement membrane, and tubular basement membranes).…”

Integration of diffraction phase microscopy and Raman imaging for label‐free morpho‐molecular assessment of live cells

“…While autofluorescence microscopy is a promising contrast‐free method but its scope is limited in visualizing or monitoring only certain metabolites in biological samples . Chemical imaging techniques based on vibrational spectroscopy, such as infra‐red (IR) and Raman microspectroscopy (RM), have engendered significant attention due to its ability to obtain abundant molecular and functional information without dye or contrast agent labeling . RM, in particular, which is based on inelastic scattering of light called Raman effect , has been widely used as a nonperturbative analytical tool to determine molecular composition .…”