Identification of fluorescent beads using a coded aperture snapshot spectral imager

Abstract

We apply a coded aperture snapshot spectral imager (CASSI) to fluorescence microscopy. CASSI records a two-dimensional (2D) spectrally filtered projection of a three-dimensional (3D) spectral data cube. We minimize a convex quadratic function with total variation (TV) constraints for data cube estimation from the 2D snapshot. We adapt the TV minimization algorithm for direct fluorescent bead identification from CASSI measurements by combining a priori knowledge of the spectra associated with each bead type. Our proposed method creates a 2D bead identity image. Simulated fluorescence CASSI measurements are used to evaluate the behavior of the algorithm. We also record real CASSI measurements of a ten bead type fluorescence scene and create a 2D bead identity map. A baseline image from filtered-array imaging system verifies CASSI's 2D bead identity map.

Multiframe image estimation for coded aperture snapshot spectral imagers

“…This instrument has been shown to effectively capture data at video rate from nonstationary scenes [6] and from beads in fluorescent microscopy [7]. The entire three-dimensional (3D) data cube is captured in a snapshot using a two-dimensional (2D) detector, where, unlike computed tomographic imaging spectrometers (CTISs) [8][9][10], the number of pixels in each channel of the data cube is equal to the total pixels of the detector.…”

Review of snapshot spectral imaging technologies

“…Ref. 155 CASSI-DD is even more compressive than CASSI-SD, so that M CASSI−DD ¼ N x N y , achieving a detector utilization equal to N w . The error can be found by a careful look at Fig.…”

Optical hyperspectral imaging in microscopy and spectroscopy - a review of data acquisition

“…The disadvantages of CASSI include its computation complexity, making it difficult to recover accurate datacubes in real time, and the requirement that the object must be sparse in the gradient domain [72]. In biomedicine, the integration of CASSI with microscopy has been demonstrated in high-resolution fluorescence imaging [73, 74]. …”