2. Spatter vs splatter Splatter: random, unorganized
Spatter: Not random, affect by gravity, predictable
Blood stain Pattern: The pattern of a stain and the quantity of blood present can be important clues to the nature of the accident or crime.
BSP Interpretation:
what does the blod tell you?

3. The Sheppard Family Story

4. Dr Sam and Marilyn Sheppard
35 blows,
28 to the head
Unknown instrument
12> inches
None of these blows were fatal
2 broken teeth
Torn fingernail.
Blood found on most of the walls along with covering a dresser.
Dr. Sam Sheppard:
Bruises
chipped teeth
fractured vertebra in his neck.
Found shirtless

5. Crime Scene pictures
Spatter ?
Sheppard Images

6. Crime Scene pictures

8. Questions
What Biological evidence will blood provide,, in the bedroom?
What information does the Blood stain evidence provide?

9. Dr Sam Sheppard July 4 1954, Reading:
HW link: main site:
Police report:

10. Do Now: What Biological evidence will blood provide,, in the bedroom?
DNA
Blood type(s)
Tool marks/ voids
Toxicological Reports
Diseases
Spatter marks (Blood trails, movements)

11. What can Spatter (Bloodstain Evidence) Evidence reveal:.

12. What can Spatter (Bloodstain Evidence) Evidence reveal:
Origin(s) of bloodstain
Position of victim & assailant
Movement of victim & assailant
Number of blows/shots
Distance of bloodstain from target
Direction from which blood impacted
Speed with which blood left its source

14. Serology The study of antigen-antibody reactions.
Tells us human vs. nonhumans
Serologists Questions
Is the sample blood?
Is the sample animal blood?
If animal blood, from what species?
If human blood, what type?
Can the sex, age, and race of the source of blood be determined?

15. History of Blood!!!
With early Transfusions =instant death due to Coagulation
, Karl Landsteiner introduced the A-B-O system
,Alexander Weiner, Rhesus monkey, Rh Factor was and over 100 factors actually must be considered when performing a transfusion.
Most people are only familiar with the Rh factor, which is technically the D antigen.
There are more than 256 antigens, and 23 blood group systems based on association with these antigens.
Us Populations: O 43%, A 42%, B 12 %, AB 3%

16. Blood Typing

17. Structure of Blood: Plasma, mostly water Cells Erythrocytes: (RBC)
Leukocytes: (WBC)
Platelets

18. Major components of Blood, Plasma:
Straw colored liquid consisting mainly (90%) of water and (7%) dissolved proteins.
Can be found outside of the circulatory system.
Also transports:
Proteins: (albumin, globulins, fibrinogen)
Salts, Glucose
Amino acids
Fatty acids,
Vitamins, Hormones,
Cellular wastes

19. DNA in BLOOD
DNA can be extracted from blood (if white blood cells which always contain a nucleus are present), and also from sperm, bone marrow, tooth pulp, and hair roots.

20. Blood, however, is commonly used in DNA testing, as per the following steps:
Blood samples are collected from the victim, defendant, and crime scene
White blood cells are separated from red blood cells
DNA is extracted from the nuclei of white blood cells
A restrictive enzyme is used to cut fragments of the DNA strand
DNA fragments are put into a bed of gel with electrodes at either end
Electric current sorts DNA fragments by length
An absorbent blotter soaks up the imprint; it is radioactively treated, and an X-ray photograph (called an autoradiograph) is produced 

21. Liquid Blood Physical properties Behaves as a projectile in motion 2
viscosity
surface tension
specific gravity
Behaves as a projectile in motion
biology, physics, maths

22. Surface Tension Resistance to penetration & separation3
Surface Tension
Resistance to penetration & separation
Surface acts to reduce surface area
Smallest SA to Volume ratio is offered by sphere

23. Dripping Blood 4 Blood trickles downwards
Blood drop grows until Wt (G) > S.T.
Single drop breaks free (teardrop shape)
Surface tension pulls in vertically
And horizontally
Shape settles into sphere (0.05 ml)
Does not break up until impact

24. . . . Drop size 5 Standard drop size 50ul (0.05ml)
Rapid bleeding gives
slightly larger drop
Shaking/movement
casts off smaller drops . . .

25. Terminal Velocity v Distance Fallen (metric)6
Terminal Velocity v Distance Fallen (metric)

26. Terminal Velocity v Distance Fallen (imperial)7
Terminal Velocity v Distance Fallen (imperial)

27. Free Falling Blood Droplets8
Free Falling Blood Droplets
0.06 ul
1.1 mm
0.5 to
0.65 m .
2.2 m/s
0.12 ul
1.32 mm
0.84
to 1 m .
3.3 m/s
0.5 ul
2.12 mm
2.4 to
3 m .
4.6 m/s .
50 ul
4.6 mm
7.5 m/s
4.2 to
5.4 m

28. Shape & Size of Bloodspot9
Shape & Size of Bloodspot
Depends mostly on nature of target surface
texture (rough or smooth)
porous or non porous
Size is related to distance fallen, provided:
standard 50 ul drop of blood
There is little change in spot diameter beyond a fall distance of 1.2 m

29. Shape & Size of Bloodspot9
Shape & Size of Bloodspot

30. Hat information does this questionable drop pelt provide?9
Hat information does this questionable drop pelt provide?

31. Height Fallen 10
Single drops of blood falling from fingertip onto smooth cardboard from various heights.
No change in diameter beyond 7 ft.
Adapted from
Introduction to Forensic Sciences,
W. Eckert, CRC, 1997

32. Effect of Target Surface11
Effect of Target Surface . . .
Spreads out smoothly
ST of spreading edge is
broken by irregular surface

33. Experiments with Falling Blood Droplets12
blood
dropper
ruler
Height
Target Surface
Fabric (theatre green)
rough paper towel
paper
whiteboard
Terazzo floor

34. Single drop of blood falling from various heights (m) onto various surfaces13
0.5 1 2 3
0.5 1 2 3
Height/Surface
smooth
floor
paper
towel
fabric

35. IMPACT ANGLE DETERMINATION
ANGLE of IMPACT is the acute angle formed between the direction of the blood drop and the plane of the surface it strikes

36. Angle of Impact 14 Adapted from Introduction to Forensic Sciences,
W. Eckert, CRC, 1997

37. Angle of Impact 14 Gravitational dense zone at lower edge 90 80 70
60
50
40
Gravitational dense zone
at lower edge
20
30
10
Adapted from
Introduction to Forensic Sciences,
W. Eckert, CRC, 1997

38. . Wave Cast-off 15 Tail of elongated stain
points in direction of travel .
Tail of wave cast-off points
back to parent drop
Parent drop
wave cast-off

39. Point of Convergence 16

40. Point of Convergence 16

41. Point of Convergence 17 5 ml blood squirted from a
syringe from height of 1 m
Point of Convergence

42. Point of Origin 1 18 Distance from point of convergence
Height above point of convergence
Origin
length
width
Angle of impact = arc sin W/L
85
60
45
30

43. Tracing Origin of Bloodspots19
Tracing Origin of Bloodspots
Point of convergence method
2 dimensional image
Point of origin method
adds 3rd dimension to image
In practice:
use of string & protractor at scene
use of computer at laboratory

44. Blood Spatter Low velocity (5 f/s, 1.5 m/s)20
Blood Spatter
Low velocity (5 f/s, 1.5 m/s)
e.g. free-falling drops, cast off from weapon
Medium velocity ( f/s, m/s)
e.g. baseball bat blows
High velocity (>100 f/s, 30 m/s)
e.g. gunshot, machinery

45. Herbert Leon MacDonell,
Laboratory of Forensic Science,
P.O. Box 1111,
Corning,
New York,
14830,
USA 21

46. Low Velocity Blood Spatter22
Low Velocity Blood Spatter
Blood source subjected to LV impact
< 5 f/s (1.5 m/s)
Spot diameter: mostly mm
some smaller, some larger
Free-falling drops (gravity only)
Cast off from fist, shoe, weapon
Dripping
Splashing
Arterial spurting

47. Cast-off from Weapon First blow causes bleeding23
Cast-off from Weapon
First blow causes bleeding
Subsequent blows contaminate weapon with blood
Blood is cast-off tangientially to arc of upswing or backswing
Pattern & intensity depends on:
type of weapon
amount of blood adhering to weapon
length of arc

48. 24
Downswing of Hammer

49. 25
Cast-off from Weapon
ceiling

50. Overhead swing with bloodied metal bar26
Overhead swing with bloodied metal bar

51. 27
Cast-off Pattern (1/2)

52. 28
Cast off Pattern (2/2) 1 2 3

53. 29
What does this tell you?

54. Cast off Pattern (2/2) ? Sequence29
Cast off Pattern (2/2) ? Sequence

55. Cast off Pattern (2/2) ? Sequence30
Cast off Pattern (2/2) ? Sequence 1
(4 spots) 2
(3 spots) 3
(2 spots)
If weapon does not pick up more blood, spatter from subsequent backswings becomes progressively less.
In practice weapon picks up more blood with each successful blow.

56. Three overhead swings with hatchet31
Three overhead swings with hatchet

57. Cast-off & medium velocity spatter32

58. Cast-off & medium velocity spatter 233
Cast-off & medium velocity spatter 2

59. Cast-off Pattern ? Object34
Cast-off Pattern ? Object

60. Cast-off Pattern from Hand35
Cast-off Pattern from Hand

61. Cast-off pattern from bloodied hand swung in front of target36
6” ruler

62. Drip Pattern 37
Free-falling drops dripping into wet blood
Large irregular central stain
Small round & oval satellite stains . . . . . . . . . . .

63. 38
Drip 1:
Blood dripping into itself from height of 1 m (8 drops)

64. Blood dripping into itself from height of 1 m (8 drops)39

65. 40
Dripping onto steps

66. Splash Pattern Volume > 1 ml41
Volume > 1 ml
Subjected to LV impact
Thrown
Tipped
Large central irregular area surrounded by elongated peripheral spatter pattern

67. 42
Splash 1
5 ml blood squirted from a syringe from a height of 1 m

68. 5 ml blood squirted from a
syringe from a height of 1 m
Splash 2 43

69. 5 ml blood squirted from a syringe from a height of 1 m44
Splash 3

70. Splash onto vertical surface45
Splash onto vertical surface
10 ml blood thrown 1 m onto
a vertical target surface
6” ruler

71. 46
Stamping in blood 1
Area seen in close-up
in next slide

72. Stamping in blood Close-up of heel area47
Stamping in blood Close-up of heel area

73. 48
Blood pool (10 drops) before stamping
Stamp 1

74. 49
Blood pool (10 drops) after stamping
Stamp 2

75. Arterial Spurt Pattern50
Arterial Spurt Pattern
Blood exiting body under arterial pressure
Large stains with downward flow on vertical surfaces
wave-form of pulsatile flow may be apparent

76. 51
spatter
Small arterial spurt
broken pottery

77. Neck incisions (scene)52

78. Medium Velocity Blood Spatter54
Medium Velocity Blood Spatter
Blood source subjected to MV impact
( f/s, m/s)
Spot diameter: mostly mm
Blows with weapon (e.g. baseball bat)

79. Medium velocity blood spatter
Medium velocity blood spatter. Point of impact 15 cm in front of vertical target surface 55
6” ruler

80. Flick 1: Blood flicked between middle finger & thumb
onto a vertical smooth surface
from a distance of 15 cm
Flick 1: 56

81. Flick 2: Blood flicked between middle finger & thumb
onto a vertical smooth surface
from a distance of 15 cm
Flick 2: 57

82. High Velocity Blood Spatter58
High Velocity Blood Spatter
Blood source subjected to HV impact
> 100 f/s, 30 m/s
Fine mist: spot size < 0.1 mm
Small mass limits spread to 1 m
!Some larger droplets reach further
Gunshot
back-spatter from entry wound
forward spatter from exit wound
High speed machinery

83. Gunshot: back& forward spatter59
Bloodstained foam held just above target surface.
Bullet passing L to R just above sheet
bullet exits foam
Bullet enters
foam
bullet
Back-spatter
on entry
Forward spatter
on exit

84. Gunshot Back Spatter Arises from entrance wound60
Arises from entrance wound
Passes back towards weapon & shooter
Seen only at close range of fire
Seen on:
inside of barrel
exterior of weapon
hand, arm, chest of shooter

85. Back spatter on steadying hand61
Back spatter on steadying hand

86. Gunshot Forward Spatter62
Gunshot Forward Spatter
Arises from exit wound
Passes forwards in same direction as shot
More copious than back-spatter
Can be seen at any range of fire
Seen on nearby surfaces, objects, persons
especially on wall behind victim

87. Forward spatter (5 ms after bullet impacted at 1000 f/s)63
bullet
blood soaked
target
2.5 cm

88. Forward spatter onto target placed 15 cm behind point of HV bullet impact (bullet passing towards screen) 1 64
6” ruler

89. Forward spatter (closer view)65

90. Forward spatter (closest view)66
5 mm

91. Wipe Patterns Object moves through a wet bloodstain67
Wipe Patterns
Object moves through a wet bloodstain
Feathered edge suggests direction

92. Transfer Patterns Wet, bloodied object contacts a secondary surface68
Wet, bloodied object contacts a secondary surface
Transfer from:
hand, fingers
shoes, weapon
hair
Transfer to:
walls, ceilings
clothing, bedding
Produces mirror-image of bloodied object

93. Transfer from hair (hair-swipe) 169

94. Transfer from hair (hair-swipe) 270

95. Flow Patterns Blood flows horizontally & vertically71
Flow Patterns
Blood flows horizontally & vertically
Altered by contours, obstacles
Often ends in pool

96. Flow pattern 72

97. Bloodspots on trousers78
Bloodspots on trousers

98. Serology
The analysis of the properties and effects of serums (blood, semen, saliva, sweat, or fecal matter) is called serology.

99. Immunoassay techniques
Looking for Drugs, toxins, antibodies
Antibodies not found in humans are synthesized
Usually inject compound with drug (that you are testing for) into an animal
Why????
Animal makes antibodies because it is a foreign substance

100. Immunoassay techniques
Enzyme Multiplied Immunoassay Technique (EMIT): Detection of drugs through a antigen-antibody reaction.
Radioimmunoassay (RIA) uses drugs that are labeled with radioactive drugs
SO DETECTION occurs with any other related forms of that drug type!!!

101. Immunoassay techniques ~~EMIT screening
Add subjects urine antibodies to detect material.
For methadone: add methadone antibodies to the urine. [Conc] of antibodies not used indicates concentration used by the drug providing a related methadone concentration

102. Immunoassay techniques, ~~EMIT screening
Marijuana's major active agent THC
Tetrahydrocannabinol (3- 4.5%)
Liquid Hashish oil 8-22 %
THC  Metabolized into THC-9-carboxlic acid
Is detectable
THC-9 in smokers urine is<1 mg (millionth of a gram)
2-5 days…10 days (sometimes)

103. Animal Responses:
Polyclonal antibodies: antibodies produced by injecting animals with a specific antigen . A series of antibodies.
A series of antibodies are produced responding to a variety of different sites on the antigen.
Monoclonal Antibodies: collection of identical antibodies that interact with a single antigen n site.
Mass produced by HYBIDOMA CELLS: spleen-cancer hybrids made in limitless supply

105. It’s Red so it must be blood, right?
When found at a CS, you must determine:
If it is blood?
Human vs animal.
How closely it can be associated with a specific individual?

106. Crime Scene analysis of blood
Confirming the stain is blood
(Presumptive tests)
Luminol
Kastle Meyer test
Leukomalachite green
Hemastix ®
Confirming the blood is HUMAN
ELISA/Precipitin test

107. Presumptive tests:
A simple test for a given substance using a reagent that changes color when mixed with the substance under investigation.
Presumptive tests are not definitive and further confirmatory tests are always required. They are used extensively in forensic science.
In general analytical chemistry, presumptive tests are often called spot tests.
The first test is simply the use of a powerful light moved across every surface of a crime scene. 
That yields possible traces for visual inspection.

108. 1. Luminol Luminol Reagent:
Tests for by production of light rather than color.
Extremely sensitive and can detect minute amounts of blood
DOES NOT interfere with subsequent DNA analysis

109. Characterization of blood stain

110. 1. Luminol Reaction RBC contain hemoglobin Mix luminol + Peroxide
The iron in homoglobin acts as a catalyst speeding up the reaction between Peroxide and luminol.
As reaction progresses, light is generated for about 30 seconds
(room should be dark)

111. 1. Luminol
sprayed across the scene because it reacts to blood by making it luminescent. 
It only takes about five seconds. 
The procedure requires that the room be considerably darkened in order to see the faint bluish glow, and the intensity of the glow increases proportionately to the amount of blood present. 
It works even with old blood or diluted stains, and can illuminate smear marks where blood has been wiped away. 
However, there is one problem with this test: luminol can destroy the properties of the blood that investigators need for further testing.  Its use is limited to proving that blood is present even if not visible.

112. 2. The Kastle-Meyer Color Test
uses a solution of phenolphthalein and hydrogen peroxide on a piece of filter paper, and when blood of any quantity is present, it turns pink. 
However, it also turns pink in the presence of potatoes or horseradish, so care must be taken at the scene.

113. Characterization of blood stain
3. Hemastix ® is a dipstick for blood
Moisten with distilled water and dipped into the sample
Positive presence of blood detected if stick turns green

114. Characterization of blood stain
5. Precipitin Tests: (10-15 years)
Serum for the precipitin test is obtained from rabbits which have produced antibodies to destroy a small quantity of human blood injected into them. 
A drop of this anti-human serum is added to suspect blood, which will precipitate its protein if it is of human origin.
Electrophoretic method: ??
Western blotting test... analysis can detect one protein in a mixture of any number of ... Western blotting tells you how much protein has accumulated in cells

115. Precipitin Tests: history
Investigators use the precipitin test to determine whether the blood is of animal or human origin. 
German biologist Paul Uhlenhuth discovered that if he injected protein from a chicken egg into a rabbit, and then mixed serum from the rabbit with egg white, the egg proteins separated from the liquid to form a cloudy substance known as precipitin. In other words, it forms an antibody. 
In the forensic test for human blood:
either a sample of the suspect blood is put into a test tube over the rabbit serum or it's used in the "gel diffusion" test, where it's placed in gel on a glass slide next to a sample of the reagent (anti-human serum). 
Passing an electric current through the glass, the protein molecules filter into the gelatin and toward each other. 
A line forms where they meet---called a precipitin line---that means the sample is human blood.

116. Precipitin Tests: history
In 1925, another blood-related discovery important to criminal investigation was made. 
Around 80 percent of the human population were found to be "secretors," which means that the specific types of antigens, proteins, antibodies, and enzyme characteristic of their blood can be found in other bodily fluids and tissues. 
In the case of a secretor, investigators can tell the blood type by examining the saliva, teardrops, skin tissue, urine, or semen.
In a rape case, for example, where the perpetrator is a secretor, potential suspects can be narrowed down through blood type analysis

117. Characterization of blood stain
5. ELISA/Precipitin Test: Human Antiserum
determines if blood is from animal or human origin.
HOW does it work?
Remember ANTIGEN-Antibody Reaction???????