In contrast to images with thicker samples (20-100 μm), the images acquired for 10 μm thick samples are highly reproducible thanks to improvements made to the image processing code and microscope setup, as explained in the Methods section.
Several notable features and caveats in the cLEDscope setup and image processing are also outlined.
Here, we perform a computational simulation of (18) F positron imaging using this modality to better understand how radioluminescence signals are formed and to assist in optimizing the experimental setup and image processing.First, the transport of charged particles through the cell and scintillator and the resulting scintillation is modeled using the GEANT4 Monte-Carlo simulation.
Differences between LSPIV-derived and reference discharge values are generally less than 20%, not systematically low or high, and not related to site parameters like channel width or depth, indicating that results are relatively insensitive to camera setup and image processing parameters typically required of LSPIV.
Together, the present experimental setup and image processing algorithm provide a standard test bench for measuring migration rates in a fully automated, robust assay which is useful for high-throughput screening in large-scale drug evaluations, where effects on migration in a 3D matrix are sought.
In order to explain the experimental setup in greater detail, the section is divided into three subsections: Apache Hadoop cluster setup, data collection and image processing and finally the classification.
This might be again due to experimental setup that heavily favours image processing algorithms or to the limited size of the test population and database.
These proof-of-concept images confirmed that the microscope setup, acquisition software, and image processing code could adequately image MMC in thin sample volumes.
(2) AdP contributed to the sample mounting, scan setup optimization for best quality, image processing method development for best quantitative analysis and in the "Methods" part of manuscript.
Given the substantially improved image quality, the increased post-acquisition image processing time is largely acceptable, though a dual illumination channel setup will be required to achieve the gains in frame rate made possible by flood illumination.
