Analysis of Cellular Autofluorescence in Touch Samples by Flow Cytometry: Implications for Front End Separation of Trace Mixture Evidence

• NCJ Number: 252379
• Author(s): M. Katherine Philpott, Christina E. Stanciu, Ye Jin Kwon, Eduardo E. Bustamante, Susan A. Greenspan, Christopher J. Ehrhardt
• Date Published: July 2017
• Length: 13 pages
• Agencies: NIJ-Sponsored
• Award: DNA Profiling of Complex Biological Mixtures using HLA-Antibody Probes and Fluorescence Activated Cell Sorting

Annotation

This study surveyed optical and biochemical variation in cell populations deposited onto a surface through touch or contact and identified specific features that may be used to distinguish and then sort cell populations from separate contributors in a trace biological mixture.

Abstract

Although researchers were not able to detect meaningful biochemical variation in touch samples deposited by different contributors with preliminary antibody surveys, they did observe distinct differences in red autofluorescence emissions (650–670 nm), with as much as a tenfold difference in mean fluorescence intensities observed between certain pairs of donors. Results indicate that the level of red autofluorescence in touch samples can be influenced by a donor’s contact with specific material prior to handling the substrate from which cells were collected. In particular, researchers observed increased red autofluorescence in cells deposited subsequent to handling laboratory gloves, plant material, and certain types of marker ink, which could be easily visualized microscopically or using flow cytometry, and persisted after hand washing. To test whether these observed optical differences could potentially be used as the basis for a cell separation workflow, a controlled two-person touch mixture was separated into two fractions via fluorescence-activated cell sorting (FACS), using gating criteria based on intensity of 650–670 nm emissions and then subjected to DNA analysis. Genetic analysis of the sorted fractions provided partial DNA profiles that were consistent with separation of individual contributors from the mixture, suggesting that variation in autofluorescence signatures, even if driven by extrinsic factors, may nonetheless be a useful means of isolating contributors to some touch mixtures. (Publisher abstract modified)