Spectrally Resolved Detector Arrays for Multiplexed Biomedical Fluorescence Imaging by Anna Siri Luthman

Author:Anna Siri Luthman
Language: eng
Format: epub, pdf
ISBN: 9783319982557
Publisher: Springer International Publishing

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taking the signal as the LS score correctly assigned to the fluorescent dye within the ROI () and the background as the LS score incorrectly assigned to the background () in the respective abundance maps. The background subtracted signal is divided by the summed variance of the signal in the ROI () and the background () weighted according to their respective fractional areas ( and ) [47].

Specular Reflectance and Glare Removal in Well Plate and Phantom Data

Ratiometric thresholding was applied to address the presence of specular reflections and glare in well plate and tissue mimicking phantom fHSI data. In these datasets, 1–5% of the image pixels were affected by specular reflections and glare. In the well plate data areas of strong specular reflections and glare probably arose from meniscus effects in the droplets containing the fluorescent dyes. In the tissue mimicking phantom the main source of specular reflections and glare appeared to be moist and wet areas on the phantom surface. In contrast, ratiometric thresholding was not required for the in vivo data since the diffusely reflective mouse skin was not as susceptible to specular reflections and glare (<0.003% of pixels).

Fig. 3.7A binary mask was calculated and used to remove the effects of specular reflection and glare from the fHSI data. The binary mask was produced by dividing the pre-processed spectral band image 12 with band 27. The resultant ratiometric image was thresholded by a z-score value of 1.5 to create a binary threshold mask. This figure shows the ROIs over the wells of a well plate after rejection of pixels suffering from specular reflections and glare

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