Forensic Bio:Sperm ID

  1. What is the average ejaculate volume?
  2. What are the components of Seminal fluid?
    • 60% Sminal vesicle fluid ( proteins and flavin)
    • 30% Prostatic fluid (AP, PSA)
    • 5% epididymis secretions
    • 5% bulburethral gland secretions
  3. The non-cellular portion of semen is known as ________
    seminal plasma
  4. The three gel forming proditens produced by the seminal vesicle are:
    • fibronectin
    • Semenogelin I
    • Semeogelin II
  5. Oligospermia
    Low sperm count of 20million sperm per ml or less!
  6. Azzospermia
    lack of sperm in semen aka aspermia
  7. Two types of Acid Phosphatase
    Semianl Acid Phosphatase (SAP) aka Prostatic Acid Phosphatase (PAP) or Prosate Specific Acid Phosphatase (PSAP) or Acid Phosphatase (AP or ACP)


    Vaginal Acid Phosphatase (VAP)
  8. SAP produced in:
    Epithelial cells in ducts of prostate gland
  9. SAP found:
    • High concentration in semen
    • Low conc. in blood of healthy males
    • Higher levels with prostate cancer
    • Male urine
  10. SAP structure
    dimeric glycoprotein
  11. Vaginal AP is present
    in vaginal secretions (vaginal drainage) =/
  12. Only difference between VAP and SAP
    electrocphertic mobility
  13. Seminal stains can exhbit AP activity for ____ if stored in a _____

    cool dry place

    Wet or dry seminal stains have been reported to maintain 100% of AP activity after 1 year stored at -20degreees C
  14. Dry seminal stains will lose HALF of their AP activity in _____ if stored at 37 deg C
    six months
  15. Wet seminal stains will lose HALF of their AP acitivity in ________ if stored at 37deg C
    2-3 months
  16. Prostate Specific Antigen (PSA) also known as:
  17. PSA is present in seminal fluid at concentrations of:
    0.5 to 2.0 mg/ml
  18. PSA is produced by
    Epithelial cells lining the ducts of the prostate gland
  19. True or False:
    PSA is confirmatory

    • some consider it to be however it is not found to be prostate specific
    • Found in low levels in: nipple aspirate fluid, breast milk, semen free vaginal swabs, female urine, tumors in female breasts, ovaries, kidneys liver, colon, espohagus, parotid, thyroid and lymphoma
    • in blood in healthy males- higher concentration in prostate cancer
  20. Approximate total length of a sperm

    head: 4-5um by 2-3um
  21. Abnormally shaped sperm accuont for ______% of total sperm in normal human males
  22. Semen Testing:

    Screening tests performed first are: _____________

    Confirmatory tests are:_____________
    Visible stains and latent stains

    immunoassays and microscopy
  23. Crystalline tests

    • The Florence Test- KI and iodine in water
    • Positive results w/ vaginal and uterine mucus from dead bodies

    • The Barberio Test-picric acide and absolute ethanol solution
    • Formed yellow spermine picrate crystals
    • Spermine later recognized to be present in many other tissue types

    • Puranen's Test- dinitronaphtholsulfonic acid or Napthol Yellow S
    • Reacts with spermine
  24. Acid Phosphatase Testing
    • Most widely used presumptive test
    • AP present in high concentrations in semen
    • highly stable
    • cheap quick and easy!

    • BCIP- forms blue precipitate
    • Diazo Red- orange/red ppt
    • Sodium Thymolphthalein Monophosphate (STMP)-blue color change
    • Fast Blue B- (Brentamine Blue) turns dark pink/violet

    Tests are qual and semi-quant
  25. The Brentamine Reaction (mechanism)
    1. a-naphthyl phosphate + Acid phosphatse ---> Free Napthol + Sodium phosphate

    2. Free napthol + colorless azo dye ---> Color change
  26. In fast blue B the free napthyl reacts with _________to produce a _____ compound
    3,3 Dimethoxybenzidine

  27. In Diazo red test the free napthyl reacts with the addtion of ______
    Fast red RC salt
  28. AP Spot the _______ and ________ are already combined to make it a one step test
    alpha-napthyl phosphate and Fast blue B
  29. Ways to ID PSA
    Radial Immunodiffusion: anit P-30 to agarose based gel

    Radial diffusion

    Rocket Immunoelectrophoresis

    Cross over electrophoresis

    Immunochromatographic tests
  30. Abacus Diagnostics PSA Cards are sensitive to ______
    0.004ug/ml or up to 1/1,000,000 semen dilutions
  31. _______ were originailly developed as a screenting test for prostate cancer
    Abacus Diagnostics PSA cards
  32. RSID-Semen is an immunochromatographic test for:
    Seminal vesicle-specific Antigen (SVSA)

    uses mouse monoclonal antibodies specific for human semenologin
  33. Seminal vesicle-Specific antigen (SVSA) is found in _______ and _______
    Semenogelin I and II
  34. ____1___ is on the test pad

    ___2____ are immobilized on the T line

    The C line has _____3_____
    1-monoclonal mouse antibodies

    2- additional mouse antibodies immobilized

    3- immobilized anti-mouse IGg acts as control
  35. ELISA is sensitive to:
    Less than 10ng/ml
  36. _____ utilizes a solid-phase double antibody sandwich assay
    Enzyme Linked immunosorbent assay (ELISA)
  37. ELISA uses:
    polyclonal or monoclonal anti-P30
  38. Dried semen stains may fluoresce under
    • Short wave and long wave UV
    • an argon laser
    • wavelenghts between 450-495nm with an orange barrier filter (interference filter of ~530nm)
  39. How ALS works:
    The emitted light has a relatively ____ intensity that is overpowered by the ______light

    To observe fluoresecne ______ must be filtered out

    excitation light

    • excitation light
  40. The single best confirmation of semen:
    Microscopic analysis!
  41. A _____ is useful in identifying intact sperm on unstained smear slides
    Phase contrast microscope
  42. Nuclear fast red will stain _____ red
    Sperm heads
  43. Picroindigocarmine will stain____ and ____ green
    Sperm tails and cytoplasm of e-cells
  44. In Xmas tree stain: ______ may also stain red
    Bacteria, yeast and WBC
  45. Hemotoxlyin ane eosin stains:
    Purple and pink

    • Heads:purple
    • Tails: pink
    • Ecell cytoplasm: pink
    • Ecell nuclie: purple
  46. Positive AP or other preliminary color test:
    Analysis of_____ gave presumptive chemical indication for the presence of AP, a component of semen
  47. Positive ALS
    Numerous fluorescing stains were observed on ____ using an ALS
  48. Positive AP and Positive P30 with Negative microscopic
    Components of semen were identified on ______, however microscopic examination failed to demonstrate presence of spermatozoa
  49. Positive AP, Negative microscopeic, negative P30
    No semen was identified on____
  50. Negative AP
    Analysis of ____failed to demosntrate presence of semen
Card Set
Forensic Bio:Sperm ID
Forensic Biology Final