Technology Commercialization and Partnerships | BSA 07-14: Fiducial Marker for Correlating Images

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Categories: biotechnology & health, electronics & instrumentation

BSA 07-14: Fiducial Marker for Correlating Images

BNL Reference Number: BSA 07-14

Patent Status: U.S. Patent Number 7,965,820 was issued on June 21, 2011

Summary

(A) Cross-section of human hair under visible light illumination. The cuticle, cortex, and medulla are indicated. (B) FTIRM reflectivity image of the Au grid. The white arrow indicates the grid intersection, used for the image correlation. (C) Au SXRF microprobe image of the Au grid. (D) FTIRM image of the protein content in the hair, generated byplotting the Amide I peak area from1600 – 1700 cm-1. (E) XRF microprobe image of copper content in the hair.(F) A red-green-blue (RGB) color image illustrating the correlation of the protein from FTIRM (blue channel) and Cu content from XRF microprobe (red channel) in the tissue. The gold grid pattern from SXRF microprobe is placed in the green channel. Scale bar for all images is 20 μm.

Synchrotron-based infrared (IR) and X-ray fluorescence (XRF) microscopes are popular tools for imaging the organic and trace metal compositions of biological materials. Fourier transform infrared microspectroscopy (FTIRM) provides chemical information on the organic components of a material while the synchrotron X-ray fluorescence (SXRF) microscopy is used to probe trace element. However to be most beneficial, it is important to combine the results from both FTIRM and SXRF imaging techniques on a single sample, which requires precise overlap of the IR and X-ray images. The invention describes a method of utilizing fiducial marker for imaging a sample substrate with both the IR and X-ray microscopes. The sample to be imaged is placed on low trace element glass slide that has a gold grid patterned on its surface. The sample can then be imaged with the SXRF and FTIRM. The gold grid pattern’s IR reflectivity image and the gold SXRF image are used as fiducial markers for spatially overlapping the FTIRM and SXRF images from the tissue.

Description

The invention describes a sample substrate containing a gold grid pattern on its surface, which can be imaged with both the IR and X-ray microscopes. The sample substrate consists of a low trace element glass slide that has a gold grid patterned on its surface, where the major and minor parts of the grid contain 25 and 12 nm gold, respectively. This grid pattern can be imaged with the infrared microscope because the reflectivity of gold differs as a function of thickness. The pattern can also be imaged with the synchrotron X-ray fluorescence (SXRF) microprobe because the gold fluorescence intensity changes with gold thickness. The tissue sample is placed on top of the patterned substrate. The grid pattern’s IR reflectivity image and the gold SXRF image are used as fiducial markers for spatially overlapping the IR and SXRF images from the tissue. Results show that IR and X-ray images can be correlated precisely, with a spatial resolution of less than one pixel (i.e., 2–3 microns).

Benefits

This technology correlates the images of SXRF and FTIRM, which can be used to diagnose and study various disease pathologies. The images of SXRF and FTIRM can be correlated with a spatial resolution of less than one pixel, i.e. 2-3 microns. In addition, the fiducial marker can be used to correlate the light path of a microscope objective with an analytical instrument or modality in situations where the light path is not visible by the analytical instrument.

Applications and Industries

By combining the images got using FTIRM and SXRF microprobe, a more complete picture of many disease states such as Alzheimer’s and exposure to environmental contaminants can be achieved.