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Supplemental Digital Content 7 (SDC7): Validation mode results
Validation of the same volume scan as shown in the manuscript figures was performed. The user selected layer 19 as the best nerve layer image (rather than layer 22, as in other analyses of this volume scan), yielding layer 9 as the wing cell layer (rather than layer 12, as in the other analyses of this volume scan).
A. Sample validation-mode qualitative results for wing cells.
In the manual-analysis image, the outer region of interest (blue outline) was defined by the software, and the cells within that region of interest were manually tagged by a user. The image on the right is a detail of an overlay that was generated by the software in validation mode, showing the qualitative congruence between the manually-tagged cells (violet centers) and the automatically detected cells (green outlines).
The software generated green, blue, and violet features. Red, orange, and yellow features were added to Figure SDC7A for clarity.
Figure SDC7A: Manual versus automatic detection of wing cells.
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B. Sample validation-mode qualitative results for dendritic immune cells.
Cells were tagged by a user in the manual-analysis image, and were detected by the software in the automatic-analysis image. The image on the right is a detail of an overlay that was generated by the software in validation mode, showing the qualitative congruence between the manually-tagged cells (blue centers) and the automatically detected cells (red outlines). The software generated blue and red features.
Violet and yellow features were added to Figure SDC7B for clarity.
Figure SDC7B: Manual versus automatic detection of immune cells.
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Table SDC7. Validation summary table for the eye shown in the manuscript figures.
Layer
Manual
Measurements Automatic Measurements Specificity and Sensitivity Count
of cells
Density per cell, µm-2
Count of cells / nerves
Density per cell,
µm-2
Average size of cell, µm2
Average length,
µm
Detected and marked, †
%
Detected and not marked, ‡
%
Marked and not detected, §
% Immune
Cells 19 11 0.00007 11 0.02352 -- 10.09 81.8 18.2 18.2
Wing
Cells 9 473 0.00567 466 0.00588 170.46 -- 94.4 5.6 0.2
Nerve
Plexus 19 -- -- 26 -- -- 250.15 -- -- --
†Features identified as cells by both the user and the software. Ideal value = 100%
‡Features identified as cells by the software but not by the user. Ideal value = 0%
§Features identified as cells by the user but not by the software. Ideal value = 0%
Immune cells:
Note that 2 of 11 features (18.8%) were identified as immune cells by the user but not by the software, and 2 of 11 features (18.8%) were identified as immune cells by the software but not by the user.
Wing cells:
The ideal value for cells detected and not marked is 0%; the actual value was 5.6% (26 of 466 cells detected by the software were not marked by the user). The ideal value for cells marked and not detected is 0%; the actual value was 0.2% (1 of 473 cells marked by the user was not detected by the software).
