Body fluid analysis

1. BODY FLUID
ANALYSIS
DR. PRATIMA

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).

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)

27. THANK YOU