Forensic semen analysis plays a vital role in identification of suspected person in rape cases. this slide consist of all the recquiried info. about semen and its analysis in forensic laboratories.
Confirmatory Test for Semen identificationParth Chuahan
Seminal fluid is a complex mixture of secretions from at least four male urogenital glands. The seminal vesicle gland contributes approximately 60% to this mixture, the prostate gland contributes approximately 30%, and the combined contribution of the epididymis and bulbourethral glands account for the remaining 10%
Confirmatory Tests for semen:
1- The Christmas Tree Stain: The most reliable confirmation for the presence of semen is the positive visual identification of sperm cells (or spermatozoa) using the Christmas tree stain.
Sample collection and preservation of biological samplesNeha Agarwal
A preliminary survey should be carried out to evaluate potential evidence. In particular, the
recognition of evidence plays a critical role in solving or prosecuting crimes. The priority of the
potential evidence at crime scenes should be assessed based on each item’s relevance to the solution
of the case. Higher priority should be assigned to evidence with probative value to the case.
For example, the evidence related to a corpus delicti is considered to be of the highest priority.
Corpus delicti is a Latin term meaning “body of crime.” In Western law, it primarily refers to the
principle that in order for an individual to be convicted, it is necessary to prove the occurrence of the crime. In a forensic investigation, it also refers to the physical evidence proving that a crime was committed
Confirmatory Test for Semen identificationParth Chuahan
Seminal fluid is a complex mixture of secretions from at least four male urogenital glands. The seminal vesicle gland contributes approximately 60% to this mixture, the prostate gland contributes approximately 30%, and the combined contribution of the epididymis and bulbourethral glands account for the remaining 10%
Confirmatory Tests for semen:
1- The Christmas Tree Stain: The most reliable confirmation for the presence of semen is the positive visual identification of sperm cells (or spermatozoa) using the Christmas tree stain.
Sample collection and preservation of biological samplesNeha Agarwal
A preliminary survey should be carried out to evaluate potential evidence. In particular, the
recognition of evidence plays a critical role in solving or prosecuting crimes. The priority of the
potential evidence at crime scenes should be assessed based on each item’s relevance to the solution
of the case. Higher priority should be assigned to evidence with probative value to the case.
For example, the evidence related to a corpus delicti is considered to be of the highest priority.
Corpus delicti is a Latin term meaning “body of crime.” In Western law, it primarily refers to the
principle that in order for an individual to be convicted, it is necessary to prove the occurrence of the crime. In a forensic investigation, it also refers to the physical evidence proving that a crime was committed
It is the beginning point for obtaining evidence which will be used by the crime scene investigator and the forensic expert
A thorough investigation of the crime scene must be completed
Crime scene is basically a scene of occurrence of crime. It is a place where a particular crime has been committed.
It is starting point for the investigator.
Blood grouping of liquid blood: forward typing and reverse typing; Blood grouping of dried blood: Lattes test, adsorption-elution, adsorption-inhibition, mixed agglutination; HLA antigens and HLA typing; Role of sero-genetic markers in individualization and paternity disputes; Pitfalls in red cell typing
It is the beginning point for obtaining evidence which will be used by the crime scene investigator and the forensic expert
A thorough investigation of the crime scene must be completed
Crime scene is basically a scene of occurrence of crime. It is a place where a particular crime has been committed.
It is starting point for the investigator.
Blood grouping of liquid blood: forward typing and reverse typing; Blood grouping of dried blood: Lattes test, adsorption-elution, adsorption-inhibition, mixed agglutination; HLA antigens and HLA typing; Role of sero-genetic markers in individualization and paternity disputes; Pitfalls in red cell typing
Semen is a “ thick, viscous, creamy, slightly yellowish or grayish” substance made up of spermatozoa — commonly known as sperm — and a fluid called seminal plasma, secret from the male reproductive organs.
The function of seminal plasma are:
To provide motility to sperm
To provide nutrition to spermatozoa
Sperm Function Tests are the keystones of evaluating functional condition of sperms. The fertility potential of a sperm will be decided not only with the number & motility but with the functional competence which is of utmost importance.
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The Roman Empire A Historical Colossus.pdfkaushalkr1407
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Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
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Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
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Instructions for Submissions thorugh G- Classroom.pptxJheel Barad
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The people of Punjab felt alienated from main stream due to denial of their just demands during a long democratic struggle since independence. As it happen all over the word, it led to militant struggle with great loss of lives of military, police and civilian personnel. Killing of Indira Gandhi and massacre of innocent Sikhs in Delhi and other India cities was also associated with this movement.
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Forensic semen analysis
1. ROHTAK
POWER POINT PRESENTATION ON – FORENSIC SEMEN ANALYSIS
SUBMITTED TO :-
FORENSIC SCIENCE DEPARTMENT
ASSIST. PROFESSOR- DR. SAPNA SHARMA
SUBMITTED BY :-
VIKAS JANGRA
ROLL NO. 2919
MSC. FORENSIC SCIENCE (F.)
SESSION – 2018-19
3. Semen, also known as seminal fluid, is an
organic fluid that may contain spermatozoa. It is
secreted by the gonads (sexual glands) and other
sexual organs
of male or hermaphroditic animals and
can fertilize female ova. In humans, seminal fluid
contains several components besides spermatozoa:
proteolytic and other enzymes as well as fructose are
elements of seminal fluid which promote the survival
of spermatozoa, and provide a azmedium through
which they can move or "swim". Semen is produced
and originates from the seminal vesicle, which is
located in the pelvis. The process that results in the
discharge of semen is called ejaculation. Semen is
also a form of genetic material. In animals, semen
has been collected for
cryoconservation. Cryoconservation of animal
genetic resources is a practice that calls for the
collection of genetic material in efforts for
conservation of a particular breed.
Fig.- Seminal Fluid
Fig. – Sperm Cells
4. The mammalian sperm cell can be divided in 4 parts:
head: it contains the nucleus with densely coiled chromatin
fibres, surrounded anteriorly by an acrosome, which contains
enzymes used for penetrating the female egg. It also contains
vacuoles.
neck: it contains one typical centriole and one atypical
centriole such as the proximal centriole like.
midpiece: it has a central filamentous core with many
mitochondria spiralled around it, used for ATP production for
the journey through the female cervix, uterus and uterine
tubes.
tail or "flagellum": it executes the lashing movements that
propel the spermatocyte.
6. During the process of ejaculation, sperm passes
through the ejaculatory ducts and mixes with fluids
from the seminal vesicles, the prostate, and
the bulbourethral glands to form the semen. The
seminal vesicles produce a yellowish viscous fluid
rich in fructose and other substances that makes up
about 70% of human semen. The prostatic secretion,
influenced by dihydrotestosterone, is a whitish
(sometimes clear), thin fluid containing proteolytic
enzymes, citric acid, acid phosphatase and
lipids.[3] The bulbourethral glands secrete a clear
secretion into the lumen of the urethra to lubricate
it.
7. Sertoli cells, which nurture and support
developing spermatocytes, secrete a fluid into
seminiferous tubules that helps transport sperm to
the genital ducts. The ductuli efferentes possess
cuboidal cells with microvilli and lysosomal granules
that modify the ductal fluid by reabsorbing some
fluid. Once the semen enters the ductus epididymis
the principal cells, which contain pinocytotic
vessels indicating fluid reabsorption, secrete
glycerophosphocholine which most likely inhibits
premature capacitation. The accessory genital ducts,
the seminal vesicle, prostate glands, and
the bulbourethral glands, produce most of the
seminal fluid.
10. B.) CHEMICAL PROPERTIES
GLAND/SITE VOLUME IN EJACULATE FEATURES
TESTIS/EPIDIDYMIS 0.15ML (5%) APPROX. 80 MILLION
SPERMATOZOA
SEMINAL VESICLE 1.5-2ML (50-65%) FRUCTOSE,
PHOSPHORYLCHOLINE
ETRGOTHINONEINE,
ASCORBIC ACID,
FLAVINS
PROSTAGLANDINS,
BICARBONATE
PROSTATE 0.6-0.9ML (20-30%) PROSTRATE, SPERMINE,
CITRIC ACID,
CHOLESTROL,
PHOSPHATEASE,
PHOSPHOLIPIDS,
FIBRINOLYSIN,
FIBINOGENASE, ZINC
ACID ,
PHOSPHATEASE,
PROSTATE SPECIFIC
11. Color: Thick, yellowish white, glairy, opalescent,
secretion having a characteristic odor known as
seminal odour.
Texture: On touch,Seminal stains are starchy.
Appearance: Garments sent for forensic
examination are usually dirty having variety of
stains, in natural light some stains are reddish
coloured, while others are brown, yellow or faint
grey in colour. These are often mixed with stains of
blood vaginal discharge, urine and semen, so as to
restrict the investigation to seminal stains only,
preliminary examination is done und
oA.) PHYSICAL OR VISUAL EXAMINATION
12. Microscopically, detection of
spermatozoa is also considered as
confirmatory evidence for the
presence of semen in a suspected
stain. In order to conduct
microscopic identification, part of
the stained garment should be
dissolved in acidulated water in
test tube.
13. The Acid phosphate test is one of the best known
and most widely employed techniques for the
purpose of identification of semen. This test is
adopted because of high acid phosphatase activity
of semen. Thus by applying this test a suspected
azoospermic or Vasectomized person who is a
suspect of sexual offence may be identified.
Stability of acid phosphatase activity in seminal
stain depends on temperature. If the stain is
exposed to sunlight The acid phosphatase activity
is slowly reduced and if the stain is heated at 60C
for 5 min. this Activity is completely destroy.
1.) ACID PHOSPHATE TEST
14. The enzyme acid phosphatase of semen
hydrolyses the calcium alpha-napthyl
phosphoric acid that Liberate as alpha
naphthol. This couple with the age forming
material to give a purple colour.
REAGENT PREPRATION
oBUFFERS
Glacial Acetic acid 1ml
Sodium acetate anhydrous 2gm
Distilled water 100ml
15. oSTEP 1 REAGENTS
oSTEP 2 REAGENTS
Buffer 50ml
Sodium alpha-naphthyl Phosphate, 0.25% (w/v) 126gm
Buffer 50ml
Naphthanil diazo blue B, 0.5% (w/v) 250gm
Step 1 Reagent and Step 2 Reagent can be made up in
bulk and aliquoted into test tubes
and frozen.
When needed, one tube of each reagent can be thawed
under warm running water for use.
16. Place a small piece (2 x 2 mm) of suspected seminal
stain
1.Stain material on Whatman filter paper or other
suitable test paper. Use proper standards and controls
including positive, negative and unstained controls.
2.Add 1-2 drops of Step 1 Reagent and allow to react for
30 seconds. (No colour should develop at this stage).
3.Add 1drop of Step 2 Reagent. Record the result after
10 seconds.
4. A positive reaction is recorded upon rapid
development of a purple colour, which is indicative of
semen.
This is not a confirmatory test for semen.
17. This method is used for the detection of choline
and spermine. In TLC, it is possible to detect
choline and Spermine simultaneously in seminal
stain. It has been possible to detect choline and
spermine in old seminal stains events after the
lapse of 5 years by this method. There are 2 types
of reagents used inTLC methods they are:-
Dragendroff’s Reagent
Potassium iodoplatinato Reagent
18. oThe fabric are classified into three groups :-
Bright (white, bright blue and pink).
Medium (blue, green, yellow, orange, red).
Dark (black, brown).
To test the excitation of the samples, we used a newly developed, very
high intensity Light source manufactured by Lumatec GmbH, Germany.
The portable system is suitable for heavy use at crime scenes because
of its low weight (approximately 6 kg), anunusually robust full metal
shell, compactness and a battery that allows 60 minutes of operation
away from an electrical outlet. A mercury lamp inside of the unit
produces a high intensity of UV (320-400nm) and visible light (400-
700nm) which was developed to detect biological stains even in
daylight. And at the end of the flexible light tube results in a beam
applicator focused on a spot or spread to a fan-like triangle.
19. Wavelengths can be chosen by eleven adjustable
filter positions (400-700,350-500,400-
500,320,400,415,440,460,490,550,570). In
combination with the eleven excitation settings,
white (>400 nm), Orange (>500 nm) and red
>590nm) goggles were used to Block the
excitation light and to visualize fluorescence more
precisely.
20. 1.) CROSS-OVER ELECTROPHORESIS
Seminal Material can be identified by demonstrating the
presence of p30, a semen specific protein. One method of
doing this is Cross-Over Electrophoresis. An extract of the
suspected stain is placed in the cathode well of a gel plate
and anti p30 is placed in the anode well. Electrophoresis is
commenced for 20 minutes at 200 volts,forcing two
components together. When the antigen (p30) meets the
antibody, a precipitin band is formed. The presence of a
precipitin band within an extract of an unknown stain
proves that the stain contains seminal material,anti-p30.
21. oPLATE PREPRATION
Clean 1” x 3” or 2” x 3” microscope slides with detergent.
Rinse and polish dry. With a diamond tipped scribe,number
the slides in the upper right-hand corner. Slides should be
consecutively numbered
oCOATING PLATE WITH GEL
On a level surface, place a pre-coated slide onto a small
inverted beaker, allowing the slide edges to be suspended.
Place one or two tubes containing 3.5 ml each of1% agarose
gel in a water bath to liquefy.1 –3.5 ml tube for a 1” x 3”
slide 2 –3.5 ml tube for a 2” x 3” slide Pour the liquefied 1%
agarose onto the slide. Surface tension and pre-coating of
slide will prevent the liquid from overflowing the edges. A
smooth level surface is formed. Allow It to cool and harden.
22. This test was discovered by Dr.Florence in the year
1886. When Florence reagent
(PotassiumIodide+Iodine+Water) is applied to the
slide it produces rhomboidal shape dark crystals
of choline periodide. Similarly, any tissue or
biological material containing sufficient high
choline concentration would give positive Florence
Test.According to Davis and Wilson, if swab is
collected within one day of sexual act,choline can
be detected. At the same time if swab is taken
after 14 hours of intercourse, there is a possibility
of negative result.
23. Barberio’s test was invented by
Barberio in the year 1905. When
the questioned stain is allowed to
react with picric acid it leads to
the formation of yellow needle
shaped spermine picrate crystals,
including the presence of seminal
stain
24. 1.) Semen analysis is an important laboratory
test and should be thought of in the same
way as any other diagnostic assay is used in
determining treatment plans for infertility.
2.) Semen analysis is very important in
assault cases as it is very Useful for
detection of semen in cloths.
3.) Isolation and identification of seminal
stain found on various suspected are asuch
as cloths, bed sheet, pillow, blanket, etc.
25. 4.) Analysis of various protein present
in semen or seminal stain.
5.) Seminal stains are examined
generally in rape cases, Sodomy,
Bestiality, Buccal coitus and in cases of
Sexual perversions. .
6.) Detection of sperm cell in Vagina
and other body parts.
7.) Feacal matter examination.