260414/ 4D–-HSI acquisition of a reference phantom/ 4DHSI Phantom

This dataset contains 4D-HSI datasets from a reference phantom containing solid dots with different mixtures of a pair of fluorophores (harmine and harmaline). Materials and Methods A dye phantom was prepared using solutions based on phosphate-buffered saline (PBS, pH 7.4) and 10% (w/v) polyvinyl alcohol (PVA), differing solely in their Ha and Hlina concentrations. Two primary stock solutions were prepared with [Hlina] = 2.34 μM and [Ha] = 2.54 μM (Sigma-Aldrich, St. Louis, MO, USA; Cat #51330 & 286044, respectively). Additionally, five intermediate mixture solutions were formulated at nominal Hlina:Ha volume ratios of 80:20, 60:40, 50:50, 40:60, and 20:80. These target volumetric ratios were approximate, as the elevated viscosity of the PVA base precluded high-precision fluidic dispensing. Aliquots of 5 μL from each solution were deposited onto glass microscope slides and allowed to dry to solidity under ambient conditions in the dark. The inclusion of PVA served as a polymer matrix that mitigated coffee-ring effect artifacts during solvent evaporation, yielding uniform solid droplets. To assess spatial uniformity across the field of view, three replicate droplets of each solution were positioned at distinct spatial coordinates on the glass substrate. Phantom description: The phantom contains dots with different relative concentrations of Harmine (Ha) and Harmaline (Hlina) as depicted in the Scheme.png file. The field of view of the images in the dataset is the one contained in the red square. Light Source WheeLED (WLS-23-A, Mightex Systems, Toronto) multi-LED light source equipped with narrow-band LEDs centered at 310, 325, 340, 365, 385, 400 and 415 nm (10 nm FWHM) coupled with a liquid light guide (Newport Corp, Irvine, CA) and a short-pass filter (TECHSPEC #84-703, Edmund Optics, Barrington, NJ), Hyperspectral Imaging (HSI) NUANCE FX hyperspectral camera (N-MSI-FX, CRi, Woburn, MA, USA) coupled with a 60mm f/2.8D lens (Perkin-Elmer, Woburn, MA). 425 nm long-pass filter (TECHSPEC #84-748, Edmund Optics, Barrington, NJ) was used to avoid spurious light in the visible range. The power of the light source at each λexc was measured using an optical power meter (model 1936-R, Newport Corp, Irvine, CA) for data preprocessing. For the beam-shape characterization, a flat fluorescent paper surface was imaged under each λexc and, independently, from four physical illumination directions (Bottom, Left, Right, Top), producing a 4D array indexed by (row, column, λexc, direction). Background cubes, lamp emission power at each wavelength and exposure time data for each cube are included for correction purposes. For each sample, several cubes were acquired under illumination with each of the wavelengths above mentioned, as described in the respective metadata files for each sample. Metadata_Paper_Bottom.csv Metadata_Paper_Left.csv Metadata_Paper_Right.csv Metadata_Paper_Top.csv Metadata_Phantom_Bottom.csv Metadata_Phantom_Left.csv Metadata_Phantom_Right.csv Metadata_Phantom_Top.csv