Nexcelom Bioscience Cellometer Auto T4, User Manual, Page: 50 / 75

Chapter 9. Cell Types
8001009 Cellometer® Auto T4 User Manual Rev C 44
Adjusting Trypan Blue Viability Parameters
Using default settings for Trypan Blue Viability Parameters provides the fastest counting speed for standard cell
types stained a solid dark blue by trypan blue. As some cell types exhibit different morphology after being stained,
it may be necessary to adjust parameters for accurate detection of the trypan blue population as indicated below.
Weak blue dead cells
To pick up weak blue dead cells, select the
Very Dim Dead Cells
check box and slowly increase
the
Enhancement
value to further refine the division
between the background and cells with low contrast
(i.e., no defined edges).
Suggested Enhancement value
is 0.60; available range is 0.04 (stained cells have
medium contrast to background) to 0.90 (stained cells
have very low contrast to background).
Note: It is a best practice to use the Very Dim Dead Cells
check box in conjunction with other Trypan Blue Viability
Parameters (such as Enhancement , Sensitivity or
Uniformity values) to avoid overcounting.
Dead cells with light blue centers
To pick up dead cells with light blue centers, slowly
increase the
Uniformity
value. Suggested value is 150;
available range is 100 (stained cells are all uniform in
color) to 255 (stained cells have non-uniform dark and
light areas).
In addition, slowly increase the Sensitivity value to
further refine image clarity. Suggested value is 1.0;
available range is .10 (stained cells are all dark) to 5.0
(stained cells are a mix of dark and light images).
Variations in sensitivity values used to pick up dead cells
with light blue centers are presented on the next page.