Forensic Blood Analysis --- 2-4-2024.pptx

Forensic Blood Analysis

1. ANALYSIS OF BLOOD SBP4502 BASIC FORENSIC Raj Cheema, PhD Department of Biomedical Sciences, FMHS, UPM [email protected]

2. ISLAM & FORENSIC SCIENCE Clear and defined guidance and framework with aims or goals intended by Islamic law for the realization of benefit to mankind Collection of proof that support each other till it form a solid and defined conclusion/evidence

3. TABLE OF CONTENTS The basics of blood (pre-class activity) Importance of blood analysis in forensic Blood spatter analysis Recap, case study and reflection (post-class activity) 01 03 02 04 BASICS SPATTER SEROLOGY SUMMARY

4. At the end of the lecture, students would be able to; Identify the fundamentals of basic serology and forensic serology . Recognize the significance of blood analysis in forensic investigations. Demonstrate an understanding of blood pattern analysis principles. LEARNING OUTCOMES

5. https://www.newsweek.com/bryan-kohbergers-lawyer-fishing-info-about-potential-jurors-1884588

6. Case #48311 Discovery of an apparent homicide 4 deceased, 1-knive, multiple blood stains discovered From the crime scene sketch, determine where the incident occurred? Height? Force? And number of injuries? REAL-CASE DEPICTION

7. 01 Comprehend the basics of forensic serology (Anatomy and Physiology) BASICS (LO 1)

8. Serology (Blood Basics) What makes up our blood? Erythrocytes (RBC) Leucocytes (WBC) Thrombocytes (Platelets) Plasma INTRODUCTION

9. Serology (Blood Facts) The average adult has about FIVE liters of blood (7-8% of BW) There’s one b illion red blood cells in two to three drops of blood For every 600 red blood cells, there are about 40 platelets and one white cell Functions – O 2 , CO 2 , fights against infection and helps heal wounds INTRODUCTION

10. Serology (Blood Types)- Mixed agglutination test There are 3 alleles for blood type: A, B, & O . Since we have 2 genes, there are 6 possible combinations. INTRODUCTION Blood Types AA or AO = Type A BB or BO = Type B OO = Type O AB = Type AB

11. PRE-CLASS DATA PutraBlast pre-class viewing quiz

12. PRE-CLASS DATA Question 3 Question 6

13. 02 Understand the importance of blood analysis in forensic SEROLOGY (LO 1 & 2)

14. Importance of blood analysis in forensic science Blood is the most common and possibly most important evidence in the solving of certain types of crimes . Nothing has been found to provide a better substitute for blood analysis. Its presence has the keen ability to link a suspect and a victim , or a suspect to a location . Blood evidence Blood samples – Can be analysed to determine blood type and DNA , which can be matched to possible suspects. Blood droplets – Can be analysed to give clues to the location of a crime , movement of a victim , and type of weapon . Blood spatter – Can be analysed to determine patterns that give investigators clues to how a crime might have happened . IMPORTANCE

15. History Dr. Eduard Piotrowski , assistant at the institute of Forensic Medicine at Poland, published first paper on Blood Spattering in 1895. It was followed by French scientist Dr. Victor Balthazard in 1939. IMPORTANCE

16. History Most profound work was done by Professor Dr. Herbert Leon MacDonell who had three publications and several books written on Blood Spattering from 1971 to 1982. IMPORTANCE

17. The serology section of a forensic laboratory may deal with any or all the following: Blood typing/grouping Characterization of unknown blood Stain patterns for crime reconstruction DNA techniques used for identification Etc. Bloodstain identification is commonly achieved by one of five methods of analysis:  Microscopic examination Chemical methods Spectrophoto -metric analysis Immunological methods DNA analysis FORENSIC SEROLOGY

18. Is it blood? Is it human blood? Can it be associated with an individual? Question 1: Question 3: Question 2: FORENSIC SEROLOGY Forensic characterization of bloodstains. Three questions that must be answered by the forensic investigator:

19. Is it blood? Question 1: FORENSIC SEROLOGY Forensic characterization of bloodstains. Three questions that must be answered by the forensic investigator: Presumptive/Screening Benzidine Test Kastle-Meyer Test Hemastix (o-tolidine, TMB) Luminol Test Others ( Guaiacum test, Aloin test, Eosin, Rhodamine, etc.) Confirmatory Takayama Test Teichmann Test Wagenaar Test RSID-Blood Others (microscopic, spectroscopic examination, electrophoresis, chromatography, RNA based assay, etc.

20. Is it human blood? Question 2: FORENSIC SEROLOGY Forensic characterization of bloodstains. Three questions that must be answered by the forensic investigator: Precipitin Test Oakley- Fulthorpe test (Double diffusion test) Rapid immunoassay/ rapid immunochromatographic tests/lateral flow assays DNA analysis (Polymerase Chain Reaction (PCR) and Short Tandem Repeat (STR) analysis)

21. Can it be associated with an individual? Question 3: FORENSIC SEROLOGY Forensic characterization of bloodstains. Three questions that must be answered by the forensic investigator: Blood typing reports / Medical Reports Police FIR/reports Etc.

22. Is it blood? Question 1: FORENSIC SEROLOGY Recap: Presumptive Benzidine Test Kastle-Meyer Test Hemastix (o-tolidine, TMB) Luminol Test Others ( Guaiacum test, Aloin test, Eosin, Rhodamine, etc.) Confirmatory Takayama Test Teichmann Test Wagenaar Test RSID-Blood Others (microscopic, spectroscopic examination, electrophoresis, chromatography, RNA based assay, etc.)

23. Forensic characterization of bloodstains. ( Presumptive Tests) In general, there are three broad categories on which the presumptive tests are divided to justify whether the given stain is blood or not? Colour Test (visible color product) - action of the peroxidase enzyme in RBCs. Chemiluminescent Test - result in the release of light as a result of the catalytic properties of blood.  Fluorescent/luminescent Test - result in the release of light from a substance that has absorbed light/energy previously .  FORENSIC SEROLOGY

24. Forensic characterization of bloodstains. ( Presumptive Colour Tests) Benzidine Test/Adler Test FORENSIC SEROLOGY

25. Forensic characterization of bloodstains. ( Presumptive Colour Tests) Kastle-Meyer (Phenolphthalein) Test FORENSIC SEROLOGY

26. FORENSIC SEROLOGY Kastle-Meyer (Phenolphthalein) Test

27. Forensic characterization of bloodstains. ( Presumptive Colour Tests) Hemastix (o-tolidine, TMB) Change ranges from orange to green to possibly blue with higher concentrations of blood Reagent on Hemastix ® is diisopropylbenzene dihydroperoxide and 3, 3’, 5, 5’‐tetramethylbenzidine (TMB) FORENSIC SEROLOGY

28. Forensic characterization of bloodstains. ( Presumptive Colour Tests) Leucomalachite green (LMG) Haemoglobin catalyses the reaction between LMG and hydrogen peroxide, converting the colourless LMG into malachite green. FORENSIC SEROLOGY

29. Forensic characterization of bloodstains. ( Presumptive C hemiluminescent  Tests) Luminol/ Bluestar Luminol reacts with blood and hydrogen peroxide, producing blue-white to yellowish-green light under very low light conditions (usually dark). FORENSIC SEROLOGY

30. Forensic characterization of bloodstains. ( Presumptive C hemiluminescent  Tests) Luminol/ Bluestar The possibility exists, however, that blood may be present in amounts too little to see with the unaided eye, or that the blood at the scene had been “cleaned up” prior to arrival of the crime scene team.  The light, or luminescence, emitted in the luminol reaction results when an oxidising agent, such as blood, catalyses the oxidation of luminol by hydrogen peroxide in a basic solution.  The reaction is not specific to blood. A follow up presumptive test, such as phenolphthalein, is typically run on potential samples prior to collection. FORENSIC SEROLOGY

31. A real case solved with Luminol https://youtu.be/k7czooqXsM4?feature=shared

32. Forensic characterization of bloodstains. ( Presumptive Fluorescent T ests) Fluorescein ( Hemascien ) Reacts with red blood cells to glow Needs an alternative light source to visualize (exposed/excited to light at the 420nm to 485nm wavelengths)  FORENSIC SEROLOGY

33. Note: How to choose the most suitable presumptive test? You need to acknowledge sensitivity, false positive, advantages and disadvantages https://forensicreader.com/presumptive-test-for-blood/

34. Forensic characterization of bloodstains. ( Confirmatory Tests) To clearly depict whether the blood sample is blood or not, a special type of test is needed to perform called  confirmatory test . These special tests are  highly specific to blood  i.e. nearly no false-positive result.  Confirmatory blood tests depends majorly on the detection of Hb molecules in RBCs. And in general scenarios, it is always done after the  presumptive test for blood  was performed.  FORENSIC SEROLOGY

35. Forensic characterization of bloodstains. ( Confirmatory Tests) There are basically  seven major types of confirmatory tests  that can be used for the confirmation of whether the possible bloodstain is actually blood or not? These are: Microscopic Examination Intact RBCs WBCs Examination (Leishman stain) Microcrystal Assay Teichmann (Hematin) Takayama Crystal Test Wagenaar Test Spectroscopic Examination Electrophoresis Method Immunoelectrophoresis RNA Based Assays Chromatography Method FORENSIC SEROLOGY Good read: https://forensicreader.com/confirmatory-test-for-blood/

36. Forensic characterization of bloodstains. ( Confirmatory Tests) Microcrystal Assay A chemical test that results in the  distinctive morphological development of microscopic crystals  as a sign of blood confirmation is called the microcrystal assay test . These microscopic heme crystals can easily be seen from a standard microscope. FORENSIC SEROLOGY Good read: https://forensicreader.com/confirmatory-test-for-blood/

37. Forensic characterization of bloodstains. ( Confirmatory Tests) RSID-Blood Rapid stain identification-blood (RSID-blood) a lateral flow immuno-chromatographic strip test format which allows the identification of human blood by detection of  glycophorin A , a red blood cell membrane antigen, using two anti-human glycophorin A (GPA) monoclonal antibodies. FORENSIC SEROLOGY Good read: https://forensicreader.com/confirmatory-test-for-blood/

38. Is it human blood? Question 2: FORENSIC SEROLOGY Forensic characterization of bloodstains. Three questions that must be answered by the forensic investigator: Precipitin Test Oakley- Fulthorpe test (double diffusion test) Rapid immunoassay/ rapid immunochromatographic tests/lateral flow assays DNA analysis (Polymerase Chain Reaction (PCR) and Short Tandem Repeat (STR) analysis)

39. FORENSIC SEROLOGY Precipitin Test (Ab-Ag Reaction)

40. FORENSIC SEROLOGY Oakley- Fulthorpe test (Double D iffusion T est) Ab is incorporated in a gel, above which a column of plain agar is placed The Ag is layered on top of the plain agar The Ag and Ab moved towards the plain agar through the intervening column of plain agar This will result in the formation of precipitate when they meet at optimum proportion.

41. Can it be associated with an individual? Question 3: FORENSIC SEROLOGY Forensic characterization of bloodstains. Three questions that must be answered by the forensic investigator: Blood typing reports Police FIR/reports Does the bloodstain appear to be related to the primary crime scene, or could it be a result of secondary transfer? Are there any additional bloodstains or related evidence nearby that require examination? Would further laboratory analysis, such as DNA profiling or serological testing, be beneficial in aiding the investigation? WILL THE BLOODSTAIN TELL A STORY?

42. Take 5

43. LET’S PLAY

44. 03 Grasp the blood spatter analysis concept SPATTER (LO 3)

45. BLOOD PATTERN ANALYSIS

46. Bloodstain spatter/pattern analysis is the examination of the shapes, location & distribution patterns of bloodstains, in order to provide an interpretation of the physical events which give rise to their origin. Information that are likely to be discovered; The direction a given droplet was traveling at the time of impact. The angle of impact. The probable distance from the target from which the droplet originated. The nature of the force involved. The nature of object used. The relative position of the suspect, victim and other related objects. The approximate numbers of blows struck during an incident. Sequencing of multiple events associated with an incident BLOOD PATTERN ANALYSIS

47. Terminologies: BLOOD PATTERN ANALYSIS

48. Blood Spatter/Pattern Analysis will delve into; Categories of Bloodstain Patterns Directionality of Bloodstain Impact Angle Determination Target Surface and Stain Shape Point of Convergence Point of Origin BLOOD PATTERN ANALYSIS

49. Categories of Bloodstain Patterns Passive Bloodstains Transfer Bloodstains BLOOD PATTERN ANALYSIS Projected Bloodstains

50. Passive Bloodstains Are drops created or formed by the force of gravity acting alone . It can be further sub-divided to include: Drops Drip patterns Pools Clots BLOOD PATTERN ANALYSIS

51. Transfer Bloodstains It is created when a wet, bloody surface comes in contact with a secondary surface . It is further sub-divided as: Contact/Transfer - an image is recognizable and may be identifiable with a particular object. Swipe/Smear - wet blood is transferred to a surface that did not have blood on it. Wipe/Smudge - a non-blood-bearing object moves through a wet bloodstain, altering the appearance of the original stain. BLOOD PATTERN ANALYSIS Contact/Transfer Swipe/Smear Wipe/Smudge

52. TEST YOURSELF Contact/Transfer Swipe/Smear Wipe/Smudge

53. Projected Bloodstains These are created when an exposed blood source is subjected to an action or force, greater than the force of gravity . It can be divided into: Arterial Spurt - Bloodstain pattern(s) resulting from blood exiting the body under pressure from a breached artery Cast-off Stains - Blood released or thrown from a blood-bearing object in motion Impact Spatter - Blood stain patterns created when a blood source receives a blow or force resulting in the random dispersion of smaller drops of blood. It can be sub-divided into low-, medium- and high velocity impact. BLOOD PATTERN ANALYSIS

54. Directionality of Bloodstain It indicates or relates the path droplet followed at the time it impacted the target BLOOD PATTERN ANALYSIS Note: The "tail" points to the direction

55. Impact Angle Determination It is the acute angle formed between the direction of the blood drop and the plane of the surface it strikes. BLOOD PATTERN ANALYSIS Angle

56. Impact Angle Determination BLOOD PATTERN ANALYSIS Circular Elliptical

57. Impact Angle Determination BLOOD PATTERN ANALYSIS

58. Impact Angle Determination BLOOD PATTERN ANALYSIS

59. Take 5

60. Impact Angle Determination BLOOD PATTERN ANALYSIS

61. Impact Angle Determination Sin θ = opposite/hypotenuse i.e. sin θ = width/length of the droplet e.g. Width = 1.5cm length = 3.0cm sin θ= Width (a) 1.5cm / Length (c) 3.0cm sin θ= 0.5 θ = 30 degrees BLOOD PATTERN ANALYSIS How about this? In a bloodstain measuring 0.04 inches wide by 4 .064 mm long. Determine the angle of impact.

62. Breakout room group activity (max 5 in a group) You are required to calculate and determine the impact angle of blood stains You will be provided with the worksheet You are given 10 minutes to complete the worksheet and You will present your data to the class GROUP ACTIVITY

63. Target Surface and Stain Shape Bloodstains can occur on a variety of surfaces. The type of surface that free falling blood strikes affects the appearance of the resulting spatter. Blood drops on a smooth surface will make a more uniform or regular circular shape. Blood drops on a rough surface will make an irregular shaped stain with rough or jagged edges. BLOOD PATTERN ANALYSIS REMINDER!

64. Point/Area of Convergence (POC/AOC) The common point, on a 2-dimensional surface (2-D) , over which the directionality of several bloodstains can be retraced. By drawing a line through the long axis of a group of bloodstains the point of convergence can be determined. Where the lines of the group of stains intersect one another the convergence point can be established. BLOOD PATTERN ANALYSIS

65. Point/Area of Convergence (POC/AOC) BLOOD PATTERN ANALYSIS

66. Point/Area of Origin (POO/AOO) The point of origin is the location from which the blood that produced the bloodstain originated. It can be constructed graphically by plotting the distance from point of convergence with their angle of impact on target surface. In 3-D. BLOOD PATTERN ANALYSIS

67. Point/Area of Origin (POO/AOO) Can be determined by these methods; Stringing method Graphic/Graph method Trigonometric method (Tangent formulae) Computer programs/ software BLOOD PATTERN ANALYSIS

68. Point/Area of Origin (POO/AOO) Trigonometric method (Tangent formulae) BLOOD PATTERN ANALYSIS

69. Point/Area of Origin (POO/AOO) Trigonometric method (Tangent formulae) Tangent of angle of impact= opposite/adjacent or Z/Y Point of origin or Z = Tan of angle of impact x Y Z = Tan of 30 degrees x 15” Z = 0.5773 x 15 = 8.66” BLOOD PATTERN ANALYSIS

70. Blood is the most common and possibly most important evidence in the solving of certain types of crimes. A drop of blood tells a thousand stories. Its COMPLEX! Its never simple……creativity is required to solve a crime. We have to look at every ANGLE! SUMMARY

71. SUMMARY

72. SUMMARY

73. Check it out! (Blood Spatter) https://youtu.be/qFR6JvVJk6o https://youtu.be/sCXfosxgyrI https://youtu.be/AHH8VItHQvk?si=Xn201zkIPbTLdvTN http://www.forensicsciencesimplified.org/blood/BloodstainPatterns.pdf REVISION LINKS/RESOURCES

74. Case #48311 Discovery of an apparent homicide 4 deceased, 1-knive, multiple blood stains discovered From the crime scene sketch, determine where the incident occurred? Height? Force? And number of injuries? REAL-CASE DEPICTION

75. POST-CLASS In this activity, you will delve into the intricacies of blood analysis at a crime scene. Your task is to provide an analysis of blood overview of these 3 case studies. Place your answer in the discussion forum in PB. Do take the time to discuss other responses too. Thank you!

76. CASE STUDY 1

77. CASE STUDY 2

78. CASE STUDY 3

79. REFERENCES General information Saferstein , Criminalistics: An Introduction to Forensic Science Siegel, Forensic Science: The Basics  Nordby , Forensic Science: An Introduction to Scientific and Investigative Techniques Presumptive Test https://forensicreader.com/presumptive-test-for-blood/ Confirmatory Test https://forensicreader.com/confirmatory-test-for-blood/ Blood Pattern Analysis https://www.crimescene-forensics.com/Crime_Scene_Forensics/Bloodstains.html

80. THANKS! DO YOU HAVE ANY QUESTIONS? [email protected] 012-5717395 Reminder Student evaluation