Quantification of confocal image analysis was done by single cell identification using the image analysis software MetaMorph (Molecular Devices Co., Tokyo, Japan), and the relative fluorescence intensity of green, red, and yellow pixels was assessed.
Quantitative confocal image analysis was done by single cell identification using Openlab software and the relative fluorescent intensity of green, red, blue, yellow, purple, light blue and white pixels was assessed.
The integrated analysis enabled cell type identification using more cells, improving the resolution of cell type heterogeneity (Supplementary Figure 24).
Specific cell type identification using immunohistochemistry or assays to quantify inflammatory genes or proteins would have provided more sensitive information about the inflammatory response.
Spanjaard, B. et al. Simultaneous lineage tracing and cell-type identification using CRISPR Cas9-induced genetiCRISPR Cas9-induced
psittaci isolation in BGM cells and subsequent identification using the IMAGEN™ direct immunofluorescence assay (DakoCytomation, Denmark), as previously described [ 24].
Chrysochromulina tobin cells were stained for identification using BODIPY 505/515 (4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene; Invitrogen, Carlsbad, CA), a neutral lipid binding fluorophore.
In addition to generating compartment-specific biomarker profiles, we aimed to develop alternative strategies circumventing the laborious 2D-PAGE for definite protein identification using limited cell numbers derived from microdissected material.
Congruence is likely to be adequate for broad-scale pattern identification using grid cells of around 1° (McInnes et al., 2009) and larger (Hurlbert & Jetz, 2007); our scale of analysis was a slightly less than 1°.
These algorithms make the functional identification of complex cells tractable, allowing guaranteed high quality identification using a much smaller set of testing stimuli and shorter time duration.
The lessened covariance, as well as smaller standard deviations, was caused by the cross-contamination of signal intensity between neighboring cells as a consequence of the cell identification scheme used in the HCS software, which does not identify individual cell boundaries correctly.
