KemoQuant™ hyperspectral imaging software allows the user to discover all independently varying spectral components associated with the chemical composition of the hyperspectral image without any prior information about the image. It uses Multivariate Curve Resolution (MCR) algorithm. This is a linear additive method that extracts the spectral components present within the image and calculates the relative quantities of each of these spectral components in each spatial pixel of the image. MCR is an iterative least squares analysis technique employing non-negativity constraints on the spectral and spatial domains of the image data. These constraints create real, interpretable pure spectral components and allow the user to make spectral assignments to the chemical composition of the pixels within the image. Because hyperspectral images can be quite large (sometimes gigabytes in size), we have licensed software implementing high-speed MCR algorithms from Sandia National Laboratories. These fast and efficient algorithms can analyze large hyperspectral images in seconds.
Once the pure spectral components are discovered, these spectral components can be stored in user-created spectral libraries for future analyses. These stored spectral components then can be used repeatedly to initialize future MCR analyses or simply be applied to the image data using Classical Least Squares (CLS) to obtain the intensities of each component within the image.
KemoQuant™ hyperspectral imaging software can be used for fluorescence, visible, VNIR, and NIR/ SWIR hyperspectral images. When working with reflectance data, the software can ratio the raw data with white and dark reference images to obtain absorbance spectra. These absorbance data can then be used with the MCR analysis.
A cross section of carrot slice was imaged through a single pass of our macrPhor™ fluorescence imaging system. The sample was excited using a 488 nm laser and emission wavelengths 500-800 nm were collected and then analyzed using KemoQuant™ software.
The KemoQuant™ algorithm identified five different spectral components associated with the sample, three of which are shown in Figure 2.
The observed components resembled Chl-a (not shown), Chl-b (red), carotenoid (not shown) and two auto-fluorescing components (green and blue). Fluorescence imaging combined with MCR analysis can provide unique insights into many other components, such as the photosynthetic pigments of plant parts, algae and other samples associated with plant growth.
We used KemoQuant™ hyperspectral imaging software to analyze VNIR hyperspectral images of potato plants grown in a greenhouse environment. Potato plants were place upon a rotating stage while the InSight whole plant hyperspectral linescan imager recorded the image of the entire plant. Figure 3 shows the false-colored image associated with three of the six spectral components associated with the plant. Figure 4 shows the spectral components used to generate the image: Chl-a (red); carotenoid (blue); and anthocyanin, which was found predominately in the stems of the potato plant (green). Whole plant imaging can provide insight into plant phenotyping and information about the photosynthetic processes of the entire plant during the growing cycle.
KemoQuant™was used to analyze hyperspectral images of agricultural soy and corn fields. MCR discovered 10 spectral components associated with the vegetation within the fields. Figure 5 shows a three-color image using four of the 10 components. Figure 6 shows the actual components. The components shown resemble Chl-a and Chl-b (shown in green), carotenoid (red), and a chlorophyll component combined with a spectral feature at 520 nm (blue). These types of analysis results can indicate the health and maturity of plants within the field. The components can be saved and used as input to the Middleton’s KemoQuant™ software for comparing different plots or charting the evolution of the component over time.
The hyperspectral images that are generated by the acquisition software can have a very large file size. If KemoQuant Analysis software is installed on the same computer that controls the hyperspectral system, the computer may also require a PCI express card to support a frame grabber or USB3 card.
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