FORENSIC SCIENCE Serology

1. FORENSIC SCIENCESerology

2. Blood Characteristics • Plasma--fluid portion of the blood (55%) • Cells (45%) • Erythrocytes--red blood cells; responsible for oxygen distribution • Leukocytes--white blood cells; responsible for “cleaning” the system of foreign invaders • Thrombocytes--platelets; responsible for blood clotting • Serum--liquid that separates from the blood when a clot is formed

3. Blood Terminology • ABO blood groups--based on having an A, B, both or none of the factors on the red blood cell • Rh factor--may be present on the red blood cell; positive if present and negative if not • Antigen--a substance found on a red blood cell • Antibody--a substance that reacts with an antigen • Agglutination--clumping of red blood cells; will result if blood types with different antigens are mixed

4. Blood Genetic Terminology • Genotype--letters that represent the traits; ie, AA, AO, BO, BB, AB and OO • Phenotype--words that describe the traits • Heterozygous--different alleles for the same trait; ie: AO, BO, AB • Homozygous--the same alleles for the trait; ie: AA, BB, OO

5. Blood Questions to be answered: • Is it blood? • Is it human blood? • Whose is it? • Determine blood type, alcohol content, drugs present • Determine the method(s) in which blood may have been deposited

6. Presumptive Tests for Blood • Kastle-Meyer color test--phenolophthalein and hydrogen peroxide are mixed together the blood’s hemoglobin will cause the formation of a deep pink color • Leucomalachite test--leucomalachite green mixed with acetic acid and distilled water; then a drop of sodium perborate will turn blood green and then darken. • Luminol test--reaction with blood results in the production of light

7. Other Tests for Blood • Microcrystalline tests--add specific chemicals to the blood in order to form characteristic crystals • Takayama • Teichmann

8. Human vs Animal • Microscopic survey • Precipitin test--blood is injected into a rabbit; antibodies are formed; rabbit’s blood is extracted as an antiserum; the antiserum is placed on sample blood. It will react with human proteins. This test is very sensitive and requires only a small amount of blood. • Characteristic Differences

9. Animal Blood Frog Blood • Larger nucleic red blood cells

10. Human Blood • Numerous non-nucleic red blood cells--5 to 6 million per mm3 • Larger but less numerous white blood cells 5 to 10,000 per mm3 • Tiny, cellular fragments 350 to 500,00 per mm3 called platelets

11. Historical Perspective Of Blood Typing Around 1900, Karl Landsteiner discovered that there are four different kinds of human blood based on the presence or absence of specific antigens found on the surface of the red blood cells. In 1940 Landsteiner and Weiner reported the discovery of the Rh factor by studying the blood of the Rhesus monkey. 85% of Caucasians, 94% of Black Americans and 99% of all Asians are Rh positive.

12. Blood Typing • A blood type has antigen A and will agglutinate with B. • B blood type has antigen B and will agglutinate with A. • AB blood type has antigen A and B and will not agglutinate with either A or B. • O blood type has neither antigen A or B and will agglutinate with either.

13. Blood Groups Can Give Blood To: Can Get Blood From Antigen Type Antibody A A B A, AB O, A B, AB O , B B B A Neither A nor B AB A and B AB A, B, O, AB Neither A nor B A, B, O, AB O A and B O

14. Blood Reactions to Antiserum REACTION Anti-A Serum Anti-B Serum BLOOD TYPE No agglutination Agglutination Agglutination No agglutination Agglutination No agglutination Agglutination No agglutination Type A Type B Type AB Type O

15. Population Distribution of Blood Types in the U. S. Type Percent O 45 A 39 B 12 AB 4

16. Typing of Dried Blood Stains Absorption-elution technique • Antiserum is placed on the blood stain. Antibodies combine with the specific antigens. • Unreacted serum is washed off the bloodstain. • Stained material is heated to 56 degrees C, breaking the antibody-antigen bond. This process is known as elution. • Known red blood cells are added. Agglutination will occur if antigens present on the added RBC’s were also originally on the stained material.

17. Differentiating Blood Types Enzymes--proteins that speed up or slow down chemical reactions. Enzymes that exist in different forms are called polymorphic and can be broken down into their separate proteins called iso-enzymes.

18. Commonly Used Blood Enzymes and Proteins • Adenosine deaminase AD • Adenylate kinase AK • Carbonic anhydrase II CA II • Erythrocyte acid phosphatase EAP • Esterase D EsD • Glucose-6-Phosphate dehydrogenase G6PD • Glyoxylase U GLO I • Group-specific component Ge • Haptoglobin Hp • Peptidase A Pep A • Phosphoglucomutase PGM • 6-Phosphogluconate dehydrogenase 6PGD • Transferrin Tf

19. PGM--phenotypes Electrophoresis can separate the protein components into these 10 iso-enzymes. - + 1- 1+ 2- 2+ 2-1- 2+1- 2-1+ 2+1+ 2+2- 1+1- Three of these are most common PGM 1+, PGM 2-1+ and PGM 2

20. EAP--phenotypes Determined by the intensity of the fluorescence. The darker the band the more it fluoresced. B C A A BA B CB C CA

21. SCENE PATTERNRECONSTRUCTION 1. Stain condition 2. Pattern 3. Distribution 4. Location 5. Directionality LAB RESULTS RECONSTRUCTION 1. Genetic marker typing 2. Age Determination 3. Source Determination 4. Race Determination 5. Sex Determination BLOOD PATTERN/SEROLOGICALRECONSTRUCTION From “Cracking Cases” by Dr. Henry C. Lee

22. Blood Stain Spatter A field of forensic study which deals with the physical properties of blood and and the patterns produced under different conditions as a result of various forces being applied to the blood. Blood, as a fluid, follows the laws of physics. It is not influenced nor affected by race, gender, or age of the one bleeding.

23. BLOOD DROPLETCharacteristics • A blood droplet will remain spherical in space until it drops onto a surface • Once a blood droplet impacts a surface, a bloodstain is formed. • A droplet falling from the same height, hitting the same surface at the same angle, will produce a stain with the same basic shape.

24. BLOOD DROPLETVolume • Is approximately 0.05 cc • Is not the same for all blood droplets--from 0.03 cc to 0.15 cc • Is directly dependent upon the surface or orifice from which it originates • The impact area is called the target.

25. CONDITIONS EFFECTINGBLOODSTAIN SHAPE • Size of the droplet • Angle of impact • Velocity at which the blood droplet left the original surface • Texture of the target surface • On clean glass or plastic--droplet will have smooth outside edges • On a rough surface--will produce scalloping on the edges

26. Questions Answered by Blood Spatter Interpretation • The distance between the target surface and the origin of blood at the time of blood shed • The point(s) of origin of the blood • Movement and direction of a person or an object • The number of blows, shots, etc. causing the bloodshed and/or the dispersal of blood.

27. Questions Answered by Blood Spatter Interpretation • Type and direction of impact that produced the bloodshed • The position of the victim and/or object during bloodshed • Movement of the victim and/or object after bloodshed

28. Bloodstain Terminology • Angle of impact--angle at which blood strikes a target surface. • Bloodstain transfer--When a bloody object comes into contact with a surface and leaves a patterned blood image on the surface. • Backspatter--blood that is directed back toward its source of energy. • Cast-off--blood that is thrown from an object in motion

29. Bloodstain Terminology • Contact stain--general term referring to bloodstains caused by contact between a wet, blood-bearing surface and a second surface which may or may not have blood on it • Transfer--image is recognizable and may be identifiable with a particular object • Swipe--wet blood is transferred to a surface which did not first have blood on it • Wipe--a non-blood bearing object moves through a wet bloodstain, altering the appearance of the original stain

30. Bloodstain Terminology • Directionality--relates to the direction a drop of blood traveled in space from its point of origin • Terminal velocity--the greatest speed to which a free falling drop of blood can accelerate in air. It is dependent upon the acceleration of gravity and the friction of the air against the blood--approximately 25:1 feet/second. • High velocity--greater than 100 feet per second; gives a fine mist appearance • Low velocity--5 feet per second or less • Medium velocity--5 to 25 feet per second.

31. Bloodstain Pattern • Terminal Velocity • Directionality • Angle of Impact

32. Blood Stain Patterns The shape of a blood stain: • Round--if it falls straight down at a 90 degree angle. • Elliptical--Blood droplet elongates as the angle decreases from 90 to 0 degrees. The angle can be determined by the following formula: width = sine of the impact angle length

33. IMPACT • The more acute the angle of impact, the more elongated the stain. • 90 degree angles are perfectly round with 80 degree angles taking on a more elliptical shape. • At about 30 degrees the stain will begin to produce a tail. • The more acute the angle, the easier it is to determine the direction of travel.

34. Blood Stains • The harder and less porous the surface, the less the blood drop will break apart. • The softer and more porous the surface, the more a blood drop will break apart. • The pointed end of the blood stain faces the direction the stain is traveling.

35. Area of Intersection The location of the blood source can be determined by drawing lines the various blood droplets to the point where they intersect. This is the blood’s origin.

36. Area of Convergence The area of convergence is the point of origin; the spot where the “blow” occurred. It is determined by drawing a line from the area of intersection straight up to where to where the angle of impact would intersect

37. CRIME SCENE What evidence can you see in this crime scene? What story does the scene tell?

38. Bring In The Dogs! The dog locates human scent. A closer look shows that the ashes have human remains and clothing. In addition, look closely at the rocks on the next slide.

40. Blood Evidence • Class evidence for blood would include blood type. If you can determine the DNA you would have individual evidence. • Blood stain patterns are considered circumstantial evidence in a court room. Experts could argue many points including direction of stains, height of the perpetrator, position of the victim, left/right hand, whether the body was moved, etc.

41. Sperm • Among the smallest and most highly specialized cells in the human body. • Has a head and a tail • Contains 23 chromosomes with the genetic material found in the head • Males release 2.5 to 6 milliliters of seminal fluid per ejaculation with approximately 100 million sperm per milliliter. Magnified 400X

42. Semen Determination of Seminal Fluid • Acid phosphatase color test • the presence of acid phosphatase, the enzyme secreted by the prostate gland into the seminal fluid, will turn purple when sodium alpha naphthylphosphate and Fast Blue B solution are placed on it. • It will also fluoresce under UV light when it comes in contact with 4-methyl umbelliferyl phosphate.

43. Semen (cont.) Determination of Seminal Fluid • Prostate Specific Antigen (PSA) or p30--unique to seminal plasma • P30 is isolated and injected into a rabbit where antibodies are produced (anti-p30) • The stain extract is place in one well of an electrophoresis plate and the anti-p30 in the opposite well. The electric is applied and the antigens and antibodies move toward each other. The formation of a precipitation line between the wells shows the presence of p30 in the sample stain. It must be seminal fluid.

44. Secretors • 80% of people are considered secretors. Their blood-type antigens are found in high concentration in their body fluids such as saliva, semen, vaginal secretions and gastric juice. If you are a secretor, you will have a higher concentration of A and B antigens than does your blood!! • With the advent of DNA, the secretor evidence is not as important as it once was.