Section 1

Section 2

Section 3

Section 4

Chapter 9 Bloodstain Pattern

Analysis

Physiology of blood

Bloodstain Pattern Analysis

(Lab Journal)

Testing if it's Blood

Spatter vs Splatter

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?

The Sheppard Family Story

Dr Sam and Marilyn Sheppard

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

Sheppard Images

Crime Scene pictures

Spatter

Crime Scene

pictures

Questions

1. What Biological evidence will blood provide,, in the bedroom?

2. What information does the Blood stain evidence provide?

Dr Sam Sheppard July 4 1954, Reading:

http://www.youtube.com/watch?v=nV4zKeGM3fc HW link: main site:

http://law2.umkc.edu/faculty/projects/ftrials/sheppard/samsheppardtrial.html

http://law2.umkc.edu/faculty/projects/ftrials/sheppard/evidence.html

Police report: http://law2.umkc.edu/faculty/projects/ftrials/sheppard/sheppardreports.html

Do Now: What Biological evidence Do Now: What Biological evidence will blood provide,, in the will blood provide,, in the bedroom?bedroom?

DNA Blood type(s) Tool marks/ voids Toxicological Reports Diseases Spatter marks (Blood trails,

movements)

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

1. .2. .3. .4. .5. .6. .7. .

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

1. Origin(s) of bloodstain2. Position of victim & assailant3. Movement of victim & assailant4. Number of blows/shots5. Distance of bloodstain from target6. Direction from which blood impacted7. Speed with which blood left its source

Section 1Section

2Section

3

Section 4

c

d

b

a

a

c

fblood types

Rhesis Factor

Times

TimesTimes

b

Turns

Turns

used to detect

d

e

Chapter 9 Bloodstain Pattern

Analysis

Physiology of blood

Bloodstain Pattern Analysis

(Lab Journal)

Testing if it's Blood

Luminol

Kastle-Meyer Test

Leukomalachite green

E.L.I.S.A.

Human vs

Blood Proteins

Antibodies

Antigen-Antibody Response

Agglutination

Enzymes

Phosphoglucomutase Esterase D,

O43%, A 42%B 12%AB 3%

Rh+, 85%Rh-, 15%

MM 30%MN 48%NN 22%

Spatter vs Splatter

Probability Rule

4 Parts of the blood

Pink

Green

cleaned

The study of antigen-antibody reactions.Tells us human vs. nonhumans

Serologists Questions1.Is the sample blood?2.Is the sample animal blood?3.If animal blood, from what species?4.If human blood, what type?5.Can the sex, age, and race of the source of blood be determined?

The study of antigen-antibody reactions.Tells us human vs. nonhumans

Serologists Questions1.Is the sample blood?2.Is the sample animal blood?3.If animal blood, from what species?4.If human blood, what type?5.Can the sex, age, and race of the source of blood be determined?

Serology

History of Blood!!!

With early Transfusions =instant death due to Coagulation1. 1901, Karl Landsteiner introduced the A-B-O system

2. 1937,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%

Structure of Blood:

1. Plasma, mostly water 2. Cells

Erythrocytes: (RBC) Leukocytes: (WBC) Platelets

Major components of Blood,Plasma:Plasma:

Straw colored liquid consisting mainly (90%) of water and (7%) dissolved proteins.

Can be found outside of the circulatory system.

Also transports:1. Proteins: (albumin, globulins, fibrinogen)2. Salts, Glucose3. Amino acids4. Fatty acids, 5. Vitamins, Hormones, 6. Cellular wastes

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.

Blood, however, is commonly used in DNA testing, as per the following steps:

1. Blood samples are collected from the victim, defendant, and crime scene

2. White blood cells are separated from red blood cells

3. DNA is extracted from the nuclei of white blood cells

4. A restrictive enzyme is used to cut fragments of the DNA strand

5. DNA fragments are put into a bed of gel with electrodes at either end

6. Electric current sorts DNA fragments by length7. An absorbent blotter soaks up the imprint; it is

radioactively treated, and an X-ray photograph (called an autoradiograph) is produced 

Liquid BloodLiquid Blood Physical properties

viscosity surface tension specific gravity

Behaves as a projectile in motion biology, physics, maths

2

Surface TensionSurface Tension

Resistance to penetration & separation Surface acts to reduce surface area

Smallest SA to Volume ratio is offered by sphere

3

Dripping BloodDripping Blood

Blood drop grows until Wt (G) > S.T.

Single drop breaks free (teardrop shape)

Surface tension pulls in vertically

Shape settles into sphere (0.05 ml)

Does not break up until impact

And horizontally

Blood trickles downwards

4

Drop size

.

Standard drop size 50ul (0.05ml)

.

Rapid bleeding gives slightly larger drop

Shaking/movementcasts off smaller drops

.

5

Terminal Velocity v Distance Fallen(metric)

Terminal velocity as a function of distance fallen (metric)

0

1

2

3

4

5

6

7

8

0 1 2 3 4 5 6 7

Fall distance (m)

Ter

min

al v

elo

city

(m

/s)

6

Terminal Velocity v Distance Fallen(imperial)

Terminal velocity as a function of distance fallen (imperial)

0

5

10

15

20

25

30

0 2 4 6 8 10 12 14 16 18 20

Distance fallen (f)

Te

rmin

al v

elo

cit

y (f

/s)

7

Free Falling Blood Droplets

.

50 ul4.6 mm

7.5 m/s

4.2 to5.4 m

0.5 ul2.12 mm

2.4 to 3 m

.4.6 m/s

0.12 ul1.32 mm

0.84 to 1 m

.3.3 m/s

0.06 ul1.1 mm

0.5 to 0.65 m

.2.2 m/s

8

Shape & Size of Shape & Size of BloodspotBloodspot

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

9

Shape & Size of Shape & Size of BloodspotBloodspot

9

Hat information does Hat information does this questionable this questionable drop pelt provide?drop pelt provide?

9

Height FallenHeight Fallen

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

10

Effect of Target Effect of Target SurfaceSurface

. .... .Spreads out smoothly ST of spreading edge is

broken by irregular surface

11

Experiments with Falling Blood DropletsExperiments with Falling Blood Droplets

blood

dropper

ruler

Terazzo floor

whiteboard

rough paper towel

paper

Fabric (theatre green)

Height

Target Surface

12

Height/Surface

Single drop of blood falling from various heights (m) onto various surfaces

smooth floor paper towel fabric

0.5

1

2

3

0.5

1

2

3

13

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

Angle of Angle of ImpactImpact

Adapted from Introduction to Forensic Sciences,W. Eckert, CRC, 1997

14

Angle Angle of of ImpacImpactt

90

10

70

2030

60 50 40

Adapted from Introduction to Forensic Sciences,W. Eckert, CRC, 1997

80

Gravitational dense zoneat lower edge

14

Wave Cast-offWave Cast-off

.Parent drop

wave cast-off

Tail of wave cast-off points back to parent drop

Tail of elongated stain points in direction of travel

15

Point of ConvergencePoint of Convergence16

Point of ConvergencePoint of Convergence16

Point of ConvergencePoint of Convergence5 ml blood squirted from a syringe from height of 1 m

17

Point of OriginPoint of Originlength

width

Angle of impact = arc sin W/L

Distance from point of convergence

Hei

ght a

bove

poi

nt o

f co

nver

genc

e

Origin

85 60 45 30

181

Tracing Origin of Tracing Origin of BloodspotsBloodspots

Point of convergence method 2 dimensional image

Point of origin methodadds 3rd dimension to image

In practice:use of string & protractor at sceneuse of computer at laboratory

19

Blood SpatterBlood Spatter

Low velocity (5 f/s, 1.5 m/s) e.g. free-falling drops, cast off from weapon

Medium velocity (25 - 100 f/s, 7.5 - 30 m/s) e.g. baseball bat blows

High velocity (>100 f/s, 30 m/s) e.g. gunshot, machinery

20

Herbert Leon MacDonell,

Laboratory of Forensic Science,P.O. Box 1111,Corning,New York,14830,USA

21

Low Velocity Blood Low Velocity Blood SpatterSpatter

Blood source subjected to LV impact < 5 f/s (1.5 m/s)

Spot diameter: mostly 4 - 8 mm some smaller, some larger

Free-falling drops (gravity only) Cast off from fist, shoe, weapon Dripping Splashing Arterial spurting

22

Cast-off from WeaponCast-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

23

Downswing of Downswing of HammerHammer

24

Cast-off from WeaponCast-off from Weaponceiling

25

Overhead swing with bloodied metal bar

26

Cast-off Pattern (1/2)Cast-off Pattern (1/2)

27

Cast off Pattern (2/2)Cast off Pattern (2/2)1

2

3

28

What does this tell What does this tell you? you?

29

Cast off Pattern (2/2) Cast off Pattern (2/2) ? Sequence? Sequence

29

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

30

Three overhead swings with hatchet

31

Cast-off & medium velocity spatter32

Cast-off & medium velocity spatter 233

Cast-off PatternCast-off Pattern? Object? Object

34

Cast-off PatternCast-off Patternfrom Handfrom Hand

35

Cast-off pattern from bloodied hand swung in front of target

6” ruler

36

.... ...

..

...

Drip PatternDrip Pattern Free-falling drops dripping into wet

blood Large irregular central stain Small round & oval satellite stains

.. .......

37

Drip 1:Drip 1:

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

38

Drip 2Drip 2

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

39

Dripping onto steps

40

Splash PatternSplash Pattern Volume > 1 ml

Subjected to LV impact Thrown Tipped

Large central irregular area surrounded by elongated peripheral spatter pattern

41

Splash 1Splash 1

5 ml blood squirted from a syringe from a height of 1 m

42

Splash 2Splash 2

5 ml blood squirted from a syringe from a height of 1 m

43

Splash 3Splash 3

5 ml blood squirted from a syringe from a height of 1 m44

Splash onto vertical surface

6” ruler

10 ml blood thrown 1 m onto a vertical target surface

45

Stamping in blood 1

Area seen in close-up in next slide

46

Stamping in blood Close-up of heel area

47

Stamp 1Stamp 1

Blood pool (10 drops) before stamping

48

Stamp 2Stamp 2

Blood pool (10 drops) after stamping49

Arterial Spurt PatternArterial Spurt Pattern Blood exiting body under arterial

pressure Large stains with downward flow

on vertical surfaces wave-form of pulsatile flow may

be apparent

50

Small arterial spurt

spatter

broken pottery

51

Neck incisions (scene)52

Medium Velocity Medium Velocity Blood SpatterBlood Spatter

Blood source subjected to MV impact (25 - 100 f/s, 7.5 - 30 m/s)

Spot diameter: mostly 1 - 4 mm Blows with weapon (e.g. baseball bat)

54

Medium velocity blood spatter.Point of impact 15 cm in front of vertical target surface

6” ruler

55

Flick 1:Flick 1:

Blood flicked between middle finger & thumb onto a vertical smooth surface from a distance of 15 cm

56

Flick 2:Flick 2:

Blood flicked between middle finger & thumb onto a vertical smooth surface from a distance of 15 cm

57

High Velocity Blood High Velocity Blood SpatterSpatter

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

58

Gunshot: back& forward spatter

Bloodstained foam held just above target surface.

Back-spatter on entry

Forward spatteron exit

bullet

Bullet passing L to R just above sheet

Bullet enters foam

bullet exits foam

59

Gunshot Back SpatterGunshot Back Spatter

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

60

Back spatter on steadying hand

61

Gunshot Forward Gunshot Forward SpatterSpatter

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

62

Forward spatter (5 ms after bullet impacted at 1000 f/s)

2.5 cm

blood soaked target

bullet

63

6” ruler

Forward spatter onto target placed 15 cm behind point of

HV bullet impact (bullet passing towards screen) 1

64

Forward spatter (closer view)65

Forward spatter (closest view)

5 mm

66

Wipe PatternsWipe Patterns Object moves through a wet

bloodstain Feathered edge suggests

direction

67

Transfer PatternsTransfer Patterns 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

68

Transfer from hair (hair-swipe) 1

69

Transfer from hair (hair-swipe) 2

70

Flow PatternsFlow Patterns Blood flows horizontally &

vertically Altered by contours, obstacles Often ends in pool

71

Flow pattern

72

Bloodspots on trousers78

Serology

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

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 animalWhy????

Animal makes antibodies because it is a foreign substance

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!!!

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

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)

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

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?

Crime Scene analysis of blood

Confirming the stain is blood

(Presumptive tests)1. Luminol2. Kastle Meyer test3. Leukomalachite

green4. Hemastix ®

Confirming the blood is HUMAN

ELISA/Precipitin test

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.

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

Characterization of blood stain

1. Luminol Reaction

1. RBC contain hemoglobin2. Mix luminol + Peroxide 3. The iron in homoglobin acts as a

catalyst speeding up the reaction between Peroxide and luminol.

4. As reaction progresses, light is generated for about 30 seconds

5. (room should be dark)

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.

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.

Characterization of blood stain

3. Hemastix ® is a dipstick for bloodMoisten with distilled water and dipped into the

samplePositive presence of blood detected if stick turns

green

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

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.

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

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???????