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BODY FLUID
ANALYSIS
DR. PRATIMA
OVERVIEW
INTRODUCTION AND BIOMEDICAL SIGNIFICANCE
FLUID COMPARTMENTS
CLASSIFICATION OF FLUIDS
SAMPLE COLLECTION [ GENERAL]
EXAMINATION OF BODY FLUIDS [ GENERAL]
BODY FLUID ANALYSIS [ INDIVIDUAL]
REFERENCES
INTRODUCTION
• Body fluids are lubricating fluids present within the body
cavities .
• Increase in the volume of the fluid in these cavities is
known as effusion .
• The commonly examined fluids are
• Urine
• CSF
• Pleural
• Pericardial
• Peritoneal and
• Synovial fluid
• Water content in body is divided into 2 compartments:
1. Extracellular fluid (ECF):
- fluid outside the cells.
» 1/3 volume of fluids in body (» 33% of total body water).
contains ions & nutrients needed for cellular life.
2. Intracellular fluid (ICF):
- fluid inside the cells.
» 2/3 volume of fluids in body (» 67% of total body water).
Fluid Compartments
» 60% of body weight
Extracellular fluid(» 1/3)
» 33% of TBW
» 20% of body wt
Interstitial fluid Plasma
Transcellular
fluid
Intracellular fluid
(» 2/3)
» 67% of TBW
» 40% of body wt
TRANSUDATE
• Malfunctioning membranes cause fluid accumulation in the
body cavities and this fluid is referred to as a transudate.
• Regulation of amount of fluid in these cavities is done by the
lymphatic system.
• Malfunctioning of membranes cause transudate formation due
to a disease process in an organ or the lymphatic system.
• Mechanism- disruption of balance between the formation and
its uptake by the lymphatic system causes fluid accumulation
in one side of the membrane.
Examples :-
• Increased hydrostatic pressure: Congestive heart failure
• Decreased oncotic pressure (decreased albumin) :liver
cirrhosis, nephrosis, and malnutrition
EXUDATES
• An exudate is a fluid with a high content of protein and cellular
debris which has escaped from blood vessels and has been
deposited in tissues.
• Cellular material-tumor cells or foreign materials such as
bacteria ,virus, parasites, fungi.
• Infection/ cancer- inflammatory response recruit large number
of white blood cells to the site.
• As a result exudate forms. Thus, cells (both leukocytes and
foreign material) and their metabolites fill the cavity in the
organ.
• EXAMPLES –
• Inflammation: Infection, infarction, hemorrhage and Tumor
Characteristics Transudate Exudate
Mechanism Filtrate of plasma due to
non –inflammatory
process with normal
vascular permeability
Increased vascular
permeability due to
inflammatory process
Appearance Clear , serous Cloudy /purulent /chylous
/hemmorrhagic
Color Straw yellow Yellow to red
Specific gravity <1.018 >1.018
Protein Low High
Clot Absent Present
Cell count Low High
Types of cells Few lymphocytes and
mesothelial cells
Neurophils and
lymphocytes
Bacteria Absent Present
LDH Low High
MODIFIED TRANSUDATE
• Moderate protein concentration: 2.5-7,5g/dl
• Moderate cellularity 1000-7000 cells/ μg
• Examples -
Cardiovascular disease
Neoplastic disease
Rupture of urinary bladder
Hepatic disease
SAMPLE COLLECTION
• FNA of effusion fluids
• Tapping
• Fluid is collected in clean , dry container under aseptic
precautions and atraumatically .
• It is collected in following 3 test tubes :-
• Chemical examination :- Flouride tube
• Microscopic examination :- EDTA tube
• Bacteriological examination : Plain tube
EXAMINATION OF BODY FLUID
• Physical examination
• Chemical examination
• Microscopic examination
PHYSICAL
EXAMINATION
Volume , color and appearance
COLOR :-
Clear and Straw colored – pleural ,pericardial and peritoneal
fluids
Blood stained – Malignancy
Turbid –high cell count or high protein content
Chylous – high lipid content due to lymphatic obstruction
CHEMICAL
EXAMINATION
• Protein examination :- helps to differentiate transudate
from exudate
• Glucose estimation :- low glucose suggests bacterial
infection , malignancy or non specific inflammation .
• Amylase :- in ascitic fluid in patients with pancreatic
lesions and etc
MICROSCOPIC EXAMINATION
CELL COUNT :- is done using neubauer chamber .
DIFFERENTIAL WBC COUNT :-
Fluid is Centrifuged and smears prepared from the sediment
And stained with Leishmans and Grams stain .
CYTOLOGICAL EXAMINATION FOR MALIGNANT CELLS :-
Smears made are fixed in absolute alcohol . Smears are
stained with H & E or Geimsa or Pap Stain.
• Adequacy: on site
• Background: necrotic, mucinous
• Cell concentration: high, low
• Cell preservation: lysis
• Inflammatory cells: which? dominant?
• Lining cells: mesothelial, epithelial
• Cells of interest: tumor cells
MICROBIOLOGICAL EXAMINATION
• Culture is done to identify the organism in cases of
effusions due to infections
DIAGNOSTIC ROLE
• It is very useful for diagnosis of premalignant and
malignant tumors, especially metastatic tumors.
• It is very useful for diagnosis of inflammatory conditions
(septic effusion, or chronic specific inflammation e.g. TB )
SEMINAL FLUID
PURPOSE OF EXAMINATION:
• INFERTILITY
• VASECTOMY FOLLOW UP
• ARTIFICIAL INSEMINATION
• MEDICOLEGAL
WHO CRITERIA FOR NORMAL SEMEN
• VOLUME –2-5ml
• PH – 7.2-7.8
• SPERM CONCENTRATION-20million/ ml
• TOTAL SPERM COUNT –40million
• MOTILITY-
50%-FORWARD PROGRESSION OR
25% OR MORE WITH RAPID LINEAR PROGRESSION
WITHIN 60 minutes of collection
• MORPHOLOGY-50%
• VIABILITY-75%
BASIC SEMEN PARAMETERS
• Colour – homogenous grey yellow opalescent ,pus,
blood -haemospermia
• Viscosity –thick mucoid viscid examined by aspiration
into pipette
• Volume - pipette or syringe
aspermia
hypo or oligospermia
hyperspermia
• PH-7.2-7.8 -
Alkaline –infection
Acidic – urine , prostatic fluid
• LIQUIFACTION TIME –20-60 MIN AFTER EJACULATION
• MICROSCOPY- sperm count
round cells
germ cells
leucocytes
motility
morphology
SPERM COUNT
• HAEMOCYTOMETER
• DILUENT-FORMALIN & SODIUM BICARBONATE
• SPERM COUNT /ml =sperms in 4 chambers x 10 x20 x1000/4
OTHERS
MARKLER COUNTING CHAMBER
HORWELL FERTILITY COUNTING CHAMBER
COUNTING BASED ON LASER PRINCIPLE
OLIGOZOOSPERMIA -<20 million
AZOOSPERMIA – M .TB,SMALL POX, MUMPS,
POLYSPERMIA-E.COLI, STREPTO FECALIS ,N.GONORRHOEAE,
CHLAMYDIA
SPERM MOTILITY
GRADING
I-RAPID PROGRESSIVE IN STRAIGHT LINE
II-SLUGGISH PROGRESSIVE
III-NON-PROGRESSIVE MOTILITY WITH SUBSTANTIAL YAW
IV -IMMOTILE
SPERM MORPHOLOGY
PAP , LEISHMAN,GIEMSA
SPERM HEAD –LENGTH:WIDTH-1.5- 1.7
TAIL DEFECTS- OFFTAIL,BIFID TAIL
HEAD ,NECK OR MIDPIECE DEFECTS-DOUBLE HEAD ,PIN HEAD,
GIANT HEAD,AMORPHOUS FORM CONSTRICTED HEAD -
TERATOZOOSPERMIA
CYTOPLASMIC DROPLETS
NORMAL –20-25% ABNORMAL FORMS
VIABILITY TESTS
Hypo-osmotic test
Dye exclusion
Supravital staining – using eosin.
BIOCHEMICAL ASSAYS
1. Zinc and citric acid in semen.
2. pH.
3. Acid phosphatase activity.
4. Fructose – low fructose: congenital absence of vasa deferentia.
5. Neutral alpha glucosidase – diagnostic of distal ductal
obstruction.
SPERM FUNCTION TESTS
• QUANTITATIVE MOTILITY –COMPUTER ASSISTED SPERM
ANALYSIS
• HAMILTON THORN MOTILITY ANALYSER
• SPERM PENETRATION ASSAY
• HEMIZONA ASSAY
• ACROSIN ASSAY
• HOS TEST
• CERVICAL MUCUS PENETRATION TEST (SIMES HUHNER
TEST)
THANK YOU

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body fluids analysis - corrected - Copy.pptx

  • 2. OVERVIEW INTRODUCTION AND BIOMEDICAL SIGNIFICANCE FLUID COMPARTMENTS CLASSIFICATION OF FLUIDS SAMPLE COLLECTION [ GENERAL] EXAMINATION OF BODY FLUIDS [ GENERAL] BODY FLUID ANALYSIS [ INDIVIDUAL] REFERENCES
  • 3. INTRODUCTION • Body fluids are lubricating fluids present within the body cavities . • Increase in the volume of the fluid in these cavities is known as effusion . • The commonly examined fluids are • Urine • CSF • Pleural • Pericardial • Peritoneal and • Synovial fluid
  • 4. • Water content in body is divided into 2 compartments: 1. Extracellular fluid (ECF): - fluid outside the cells. » 1/3 volume of fluids in body (» 33% of total body water). contains ions & nutrients needed for cellular life. 2. Intracellular fluid (ICF): - fluid inside the cells. » 2/3 volume of fluids in body (» 67% of total body water).
  • 5.
  • 6. Fluid Compartments » 60% of body weight Extracellular fluid(» 1/3) » 33% of TBW » 20% of body wt Interstitial fluid Plasma Transcellular fluid Intracellular fluid (» 2/3) » 67% of TBW » 40% of body wt
  • 7. TRANSUDATE • Malfunctioning membranes cause fluid accumulation in the body cavities and this fluid is referred to as a transudate. • Regulation of amount of fluid in these cavities is done by the lymphatic system. • Malfunctioning of membranes cause transudate formation due to a disease process in an organ or the lymphatic system. • Mechanism- disruption of balance between the formation and its uptake by the lymphatic system causes fluid accumulation in one side of the membrane. Examples :- • Increased hydrostatic pressure: Congestive heart failure • Decreased oncotic pressure (decreased albumin) :liver cirrhosis, nephrosis, and malnutrition
  • 8. EXUDATES • An exudate is a fluid with a high content of protein and cellular debris which has escaped from blood vessels and has been deposited in tissues. • Cellular material-tumor cells or foreign materials such as bacteria ,virus, parasites, fungi. • Infection/ cancer- inflammatory response recruit large number of white blood cells to the site. • As a result exudate forms. Thus, cells (both leukocytes and foreign material) and their metabolites fill the cavity in the organ. • EXAMPLES – • Inflammation: Infection, infarction, hemorrhage and Tumor
  • 9. Characteristics Transudate Exudate Mechanism Filtrate of plasma due to non –inflammatory process with normal vascular permeability Increased vascular permeability due to inflammatory process Appearance Clear , serous Cloudy /purulent /chylous /hemmorrhagic Color Straw yellow Yellow to red Specific gravity <1.018 >1.018 Protein Low High Clot Absent Present Cell count Low High Types of cells Few lymphocytes and mesothelial cells Neurophils and lymphocytes Bacteria Absent Present LDH Low High
  • 10. MODIFIED TRANSUDATE • Moderate protein concentration: 2.5-7,5g/dl • Moderate cellularity 1000-7000 cells/ μg • Examples - Cardiovascular disease Neoplastic disease Rupture of urinary bladder Hepatic disease
  • 11. SAMPLE COLLECTION • FNA of effusion fluids • Tapping • Fluid is collected in clean , dry container under aseptic precautions and atraumatically . • It is collected in following 3 test tubes :- • Chemical examination :- Flouride tube • Microscopic examination :- EDTA tube • Bacteriological examination : Plain tube
  • 12. EXAMINATION OF BODY FLUID • Physical examination • Chemical examination • Microscopic examination
  • 13. PHYSICAL EXAMINATION Volume , color and appearance COLOR :- Clear and Straw colored – pleural ,pericardial and peritoneal fluids Blood stained – Malignancy Turbid –high cell count or high protein content Chylous – high lipid content due to lymphatic obstruction
  • 14. CHEMICAL EXAMINATION • Protein examination :- helps to differentiate transudate from exudate • Glucose estimation :- low glucose suggests bacterial infection , malignancy or non specific inflammation . • Amylase :- in ascitic fluid in patients with pancreatic lesions and etc
  • 15. MICROSCOPIC EXAMINATION CELL COUNT :- is done using neubauer chamber . DIFFERENTIAL WBC COUNT :- Fluid is Centrifuged and smears prepared from the sediment And stained with Leishmans and Grams stain . CYTOLOGICAL EXAMINATION FOR MALIGNANT CELLS :- Smears made are fixed in absolute alcohol . Smears are stained with H & E or Geimsa or Pap Stain.
  • 16. • Adequacy: on site • Background: necrotic, mucinous • Cell concentration: high, low • Cell preservation: lysis • Inflammatory cells: which? dominant? • Lining cells: mesothelial, epithelial • Cells of interest: tumor cells MICROBIOLOGICAL EXAMINATION • Culture is done to identify the organism in cases of effusions due to infections
  • 17. DIAGNOSTIC ROLE • It is very useful for diagnosis of premalignant and malignant tumors, especially metastatic tumors. • It is very useful for diagnosis of inflammatory conditions (septic effusion, or chronic specific inflammation e.g. TB )
  • 18. SEMINAL FLUID PURPOSE OF EXAMINATION: • INFERTILITY • VASECTOMY FOLLOW UP • ARTIFICIAL INSEMINATION • MEDICOLEGAL
  • 19. WHO CRITERIA FOR NORMAL SEMEN • VOLUME –2-5ml • PH – 7.2-7.8 • SPERM CONCENTRATION-20million/ ml • TOTAL SPERM COUNT –40million • MOTILITY- 50%-FORWARD PROGRESSION OR 25% OR MORE WITH RAPID LINEAR PROGRESSION WITHIN 60 minutes of collection • MORPHOLOGY-50% • VIABILITY-75%
  • 20. BASIC SEMEN PARAMETERS • Colour – homogenous grey yellow opalescent ,pus, blood -haemospermia • Viscosity –thick mucoid viscid examined by aspiration into pipette • Volume - pipette or syringe aspermia hypo or oligospermia hyperspermia
  • 21. • PH-7.2-7.8 - Alkaline –infection Acidic – urine , prostatic fluid • LIQUIFACTION TIME –20-60 MIN AFTER EJACULATION • MICROSCOPY- sperm count round cells germ cells leucocytes motility morphology
  • 22. SPERM COUNT • HAEMOCYTOMETER • DILUENT-FORMALIN & SODIUM BICARBONATE • SPERM COUNT /ml =sperms in 4 chambers x 10 x20 x1000/4 OTHERS MARKLER COUNTING CHAMBER HORWELL FERTILITY COUNTING CHAMBER COUNTING BASED ON LASER PRINCIPLE OLIGOZOOSPERMIA -<20 million AZOOSPERMIA – M .TB,SMALL POX, MUMPS, POLYSPERMIA-E.COLI, STREPTO FECALIS ,N.GONORRHOEAE, CHLAMYDIA
  • 23. SPERM MOTILITY GRADING I-RAPID PROGRESSIVE IN STRAIGHT LINE II-SLUGGISH PROGRESSIVE III-NON-PROGRESSIVE MOTILITY WITH SUBSTANTIAL YAW IV -IMMOTILE
  • 24. SPERM MORPHOLOGY PAP , LEISHMAN,GIEMSA SPERM HEAD –LENGTH:WIDTH-1.5- 1.7 TAIL DEFECTS- OFFTAIL,BIFID TAIL HEAD ,NECK OR MIDPIECE DEFECTS-DOUBLE HEAD ,PIN HEAD, GIANT HEAD,AMORPHOUS FORM CONSTRICTED HEAD - TERATOZOOSPERMIA CYTOPLASMIC DROPLETS NORMAL –20-25% ABNORMAL FORMS
  • 25. VIABILITY TESTS Hypo-osmotic test Dye exclusion Supravital staining – using eosin. BIOCHEMICAL ASSAYS 1. Zinc and citric acid in semen. 2. pH. 3. Acid phosphatase activity. 4. Fructose – low fructose: congenital absence of vasa deferentia. 5. Neutral alpha glucosidase – diagnostic of distal ductal obstruction.
  • 26. SPERM FUNCTION TESTS • QUANTITATIVE MOTILITY –COMPUTER ASSISTED SPERM ANALYSIS • HAMILTON THORN MOTILITY ANALYSER • SPERM PENETRATION ASSAY • HEMIZONA ASSAY • ACROSIN ASSAY • HOS TEST • CERVICAL MUCUS PENETRATION TEST (SIMES HUHNER TEST)