The RAS design involves many aspects, both physical and biological: (1) a desired turnover, (2) fingerling arrival frequency, (3) number of fingerlings per batch, (4) number of days in a growth phase, (4) timing of grading and sorting, based on (5) fish growth rate, and (6) number of culture tanks.
Transuded cells were isolated by fluorescence activated cell sorting based on GFP expression.
These variable and stable subpopulations consisted of OTUs from the top and bottom deciles after sorting based on variance of fluorescence intensity over the 3 days.
As shown in Table 1, sorting based on either FSC-H or SSC-H significantly reduced the quality of the size separation for all cell types, as did utilizing SSC-W as the gating parameter.
In the literature protocols utilizing culture methods, sorting based on acetylated low-density lipoprotein (ac-LDL) uptake or expression of CD31 have been described for isolating EC from non-muscle tissues [14] [17].
Sorting based on FSC-W (Figure 2) clearly improved the size separation for both L1210 and FL5.12 (by >15%), with percent overlap reaching the lowest values of 8.5% and 15% respectively, but only minimally improved on the size separation achieved by SSC-A in HL60 cells.
This viewer also allows row sorting based on any column criteria.
Lymphoid and myeloid cell subsets were isolated by FACS sorting based on their surface-marker expression profiles as described above.
The FPD can be determined quite rapidly and therefore the possibly of sorting based on this value is an important future direction.
Previously published study of the human fibroblasts and epithelial cells sorting based on the cell tracker staining showed similar results [ 25].
Sorting based on expression of CD45RB and CD25 facilitates sub-fractionation of CD4+ T cells into naYve (CD45RBhi), memory (CD45RBloCD25−) and regulatory (CD45RBloCD25+) populations 37.
