Separation of uncompromised whole blood mixtures for single source STR profiling using fluorescently-labeled human leukocyte antigen (HLA) probes and fluorescence activated cell sorting (FACS)

Fluorescence- and magnetic-activated cell sorting strategies to separate spermatozoa involving plural contributors from biological mixtures for human identification

“…In recent studies, flow cytometry has also been successfully applied to identify and separate sperm cells from vaginal epithelial cell mixtures in forensic science922. However, only scarce data has been published applying this approach to mixed samples from two or more donors with the same gender111223. We, for the first time, aimed to identify single donor’s genotype from mixed sperm cells involving plural contributors by FACS.…”

“…There has been limited studies using FACS (fluorescent-activated cell sorting) to separate cells from forensic mixtures, including sperm cells and epithelial cells mixtures9, and non-compromised blood and saliva mixtures11. More recently, Dean and colleagues exploits the intrinsic immunological variation among individuals to physically separate single donor cells in uncompromised whole blood mixtures by means of HLA antibody probes coupled to FACS12. In this study, we, for the first time, tested the feasibility of applying this technique for the isolation of single donor cells from mixed sperm cells involving plural contributors based on their ABO blood types.…”

Analysis of red autofluorescence (650-670nm) in epidermal cell populations and its potential for distinguishing contributors to 'touch' biological samples

“…Differentiating cell populations from individual contributors in a biological mixture before DNA analysis is a potential way to overcome this issue. While strategies exist to selectively label cell populations from distinct contributors based on their immunochemistry and then physically isolate cells from the mixture prior to DNA profiling 2– 4 , there is a dearth of studies demonstrating cell separation techniques on touch samples. This is likely due to the fact that cell populations in these samples mostly, if not entirely, consist of fully differentiated keratinocytes which have limited reactivity to common molecular probes used to target surface antigens 5, 6 .…”

Flow cytometry dataset for cells collected from touched surfaces

“…non-degraded) forensic mixture sample ( Dean et al , 2015; Schoell et al , 1999; Verdon et al , 2015). However, application to ‘touch’ or trace epithelial cell mixtures remains a challenge since many cell surface features are lost or obscured during the process of keratinocyte differentiation, leaving few biochemical or structural features in shed corneocytes that vary between individual contributors.…”

“…We also investigated the capacity of touch epithelial cells to bind to two different classes of antibody probes: Human Leukocyte Antigen (HLA), which has been successfully utilized to separate uncompromised mixtures of blood and other bodily fluids (e.g. Dean et al , 2015) and Cytokeratin (CK), which is known to be a dominant component of epidermal cells. This dataset includes samples that were collected and analyzed immediately after deposition (i.e.…”