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High performance image mapping spectrometer (IMS) for snapshot hyperspectral imaging applications

Michal E. Pawlowski, Jason G. Dwight, Thuc-Uyen Nguyen, and Tomasz S. Tkaczyk

Open Access Open Access

Abstract

A high performance, snapshot Image Mapping Spectrometer was developed that provides fast image acquisition (100 Hz) of 16 bit hyperspectral data cubes (210x210x46) over a spectral range of 515-842 nm. Essential details of the opto-mechanical design are presented. Spectral accuracy, precision, and image reconstruction metrics such as resolution are discussed. Fluorescently stained cell samples were used to directly compare the data obtained using newly developed and the reference image mapping spectrometer. Additional experimental results are provided to demonstrate the abilities of the new spectrometer to acquire highly-resolved, motion-artifact-free hyperspectral images at high temporal sampling rates.

© 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement

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Supplementary Material (2)

NameDescription
Visualization 1       Visualization 1
Visualization 2       Visualization 2

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Figures (9)
Fig. 1
Fig. 1 Opto-mechanical schematic of the IMS system (a). Three-dimensional model of the complete system, with chassis walls drawn in semi-transparent mode (b). Photograph of assembled prototype with top cover removed (c).

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Fig. 2
Fig. 2 Optical model of the re-imaging system of the IMS system limited to the prism-lenslet –detector assembly (a). Nominal spot diagrams for axial (Obj: 0.0, 0.0 mm) and diagonal edge field points (Obj: 9.1, 13.5 mm) for 515 and 842 nm (b).

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Fig. 3
Fig. 3 Exploded view model of the lenslet assembly (a). Cross-section through the lenslet assembly with critical dimensions and elements indicated (b). Photograph of complete assembly with one-cent coin for size comparison (c).

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Fig. 4
Fig. 4 Reconstructed images of the 1951 USAF resolution target (a) and Ronchi ruling (c). Designed (red line) and measured (black line) system dispersion expressed in lateral shift measured in pixel/nm (c).

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Fig. 5
Fig. 5 Spectral plots at three selected emission lines of the Fianium laser. Data for nominal 540, 650 and 730 nm laser lines are presented in figures (a), (b) and (c) respectively. Red dots represent HS-IMS data averaged over the field of view, the continuous green line is for Ocean Optics spectrometer raw data, and the blue line is for HS-IMS data interpolated with a cubic spline curve. Insets in the figures depict the area in the vicinity of a laser line peak.

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Fig. 6
Fig. 6 Variance vs. mean intensity signal as measured for the PCO.EDGE 5.5 camera. Raw data points are marked with red dots; the red line show raw signal linearly interpolated between sampling points and the green line depicts a first degree polynomial fitted to the linear part of the data set. Inset in the right bottom of the plot depicts magnified region of correlogram, for which average signal was below 8000 counts.

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Fig. 7
Fig. 7 Pseudo-colored image of a fluorescently stained sample of a bovine pulmonary artery endothelial cell sample recorded at 537 nm for a newly developed (HS-IMS) (a) and reference (P-IMS) spectrometer (b). Intensity cross-sections though both data sets in the direction marked on (a) and (b) by white lines are given in (c) and (d). HS-IMS and P-IMS signals are drawn in blue and orange lines respectively; individually scaled signals are shown in (c) and shared y-axis scale for both plots is provided in (d). Fluorescent signal from the BPAEC sample recorded at 617 nm by the newly developed (e) and reference spectrometer (f). Intensity cross-sections in the direction marked on (e) and (f) by white lines are given in (g) and (h). Individually scaled plots with left and right hand side scale for HS-IMS and P-IMS respectively (g). Plots from (g) with single y axis scale (h). Red rectangle marks area of the field of view area shared by both systems. Axis scales are given in physical units of length in the sample space.

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Fig. 8
Fig. 8 Picture of red ping pong balls dyed with Spectre 300 infrared dye (top row) and un-painted (bottom row) (a). Arbitrary selected frame, recorded at time t = 0.78 s with two ping-pong balls in free fall (b). Gray scale image sub-titled “VIS” shows hyperspectral data cube integrated over 513-605 nm and 2D image “IR” shows hyperspectral data cube integrated in range 610-777 nm. Spectral cross-sections taken at points marked with A and B through dyed (dash dotted line) and un-dyed (continuous line) (c). Movie compiled from a series of consecutive hyperspectral data cubes is given in Visualization 1. Two dimensional representations of hyperspectral data cube were flat field corrected and digitally post-processed with a 5x5 median filter for visualization purposes. Presented spectral cross-sections were averaged over a 3x3 area.

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Fig. 9
Fig. 9 Images and spectral profiles of a halogen bulb emitter acquired during a power on/off cycle. A pseudo-colored, panchromatic image of the halogen bulb emitter in an off state acquired at time t = 0 (a). A panchromatic image of a tungsten emitter in an on state, acquired at time t = 2.75 s (b). Spectral intensity plots taken at t = 0.9 s, t = 1.7 s and t = 3.07 s scaled (to the detector’s dynamic range) and normalized are given in (c) and (d) respectively. Time laps recording of the halogen bulb during an on/off cycle is given in Visualization 2.

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Tables (2)

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Table 1 Spectral accuracy and precision of IMS system as measured against Ocean Optics USB2G38810. Ocean Optics λOO –central wavelength as measured by Ocean Optics USB2G38810; Ocean Optics FWHM – full width at half maximum of laser line as measured by the Ocean Optics spectrometer; IMS λIMS – central wavelength of laser line as measured by the IMS system; IMS ± σ – standard deviation of measured central wavelength of the laser line as measured by the IMS;IMS FWHM – full width at half maximum of the laser line as measured by the IMS system; Δλ – difference between IMS and Ocean Optics central wavelength.

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Table 2 Quantitative comparison of fluorescent signal level between P-IMS and HS-IMS systems. Three arbitrary selected microspheres (FluoSpheres, #F8841, Thermo-Fisher) were imaged at 30, 20, 10 and 5 ms exposure times respectively. Averaged (168 images) intensity values integrated over spatial and spectral voxels of selected beads for both systems are given in columns ItotHS-IMS and ItotM-IMS. Count of photo-electrons recorded by HS-IMS and P-IMS systems over spatio-spectral voxels of each bead is given in Ie-HS-IMS and Ie-M-IMS respectively. Please note that signal recorded by P-IMS was multiplied by 0.5 to account for 2x binning in y direction.

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