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LMD of Sperm from a Cell Mixture

LMD of Sperm from a Cell Mixture. Why is cell separation important?. DNA mixtures are frequently encountered in forensic evidence particularly in sexual assault crimes. Identify individuals involved with a crime

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LMD of Sperm from a Cell Mixture

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  1. LMD of Sperm from a Cell Mixture

  2. Why is cell separation important? • DNA mixtures are frequently encountered in forensic evidence particularly in sexual assault crimes. • Identify individuals involved with a crime • Genotype mixture interpretation and statistical analysis can be challenging and time consuming • Separation of different cell types can provide cell fractions from individuals involved in a crime.

  3. Separation Methods • Preferential Lysis (differential extraction) • Flow Cytometry • Microchip separation • Membrane Filtration • Magnetic Antibodies • Y- chromosome (non-physical separation) • Laser Microdissection

  4. LMD Technology Microscope Objective Slide  Specimen Tube 

  5. Sperm & Epithelial Cell Mixture • Technical Issues of Sperm ID under LMD using brightfield conditions • PEN film architecture • Stain is necessary • It helps to adjust focal plane • May be resolved by new EP “Director Slides” • High density of epithelial cells • May need to dilute pellet over larger area • No cover slip • Looking through 1mm slide • Set correction collar objective

  6. Identification of sperm cells Leica AS LMD instrument (older version of software shown)

  7. After Laser Microdissection Sperm cuttings in tube

  8. Epithelial Cell Recovery Epithelial cell in collection tube

  9. Current LMD Collection Software

  10. Warning! Laser in Use

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