This document summarizes histological and cytological specimens. It describes tissue samples obtained from biopsies or autopsies that can be either histological (muscular) or cytological (liquid). Cytological specimens include bodily fluids like urine, cerebrospinal fluid, joint fluid, endoscopy samples, sputum, and other fluids. The procedures for processing and staining samples of each fluid type are provided. Semen is also discussed as a cytological specimen, including normal parameters for analysis and staining techniques.

1. HISTOLOGICAL AND
CYTOLOGICAL SPECIMENS

2. HISTOLOGICAL SPECIMENS
• It includes all kinds of the tissue removed from the body e.g.
breast, uterus, liver, prostate etc.
• The tissue can be obtained either by biopsy or autopsy
process.
• Biopsy is a process of removing a tissue from patient for
diagnostic examination while autopsy is process where by a
tissue taken from a dead body.
• The tissue may either be cytological (in liquid form) or
histological ( in muscular form)

3. CYTOLOGICAL SPECIMENS
• It includes different types of the fluid found in the body.eg
• 1.URINE
• 2.CSF(cerebral spinal fluid)
• 3.JOINT FLUID
• 4.ENDOSCOPY SPECIMENS
• 5.SPUTUM
• 6.OTHER BODY FLUIDS e.g. pleural fluid, peritoneal fluid and
pericardium.
• 7.SEMEN

4. 1.URINE
• Urine assist in the detection of malignancy arising from the
urinary system.
PROCEDURE
1.Specimen is centrifuged, supernatant poured out and the
deposit is smeared on the clean dry slides and fixed
immediately in ether/ 95% alcohol for at least 15min.
a)-The smear(wet) is stained by papanicolauous method and
mounted
b)-Air dried smear are stained by Rowmanowsky stains e.g.
-Giemsa
-Field
-Leishman

5. 2.CSF
• CEREBRAL SPINAL FLUID:Is commonly obtained by lumber
puncher.
• PROCEDUDE
1)Centrifuge immediately after collection
2)Pour the supernatant and smear the deposit on the clean glass
slides and fix immediately in ether/95% alcohol for at least
15min and stain by
- Papanicolauous (wet)
-Air dry and stain with Rowmanowsky stain

6. 3.JOINT FLUID
• Synovial fluid from joints may also be processed and
examined for infectious pathogens or malignancy.
• Specimen is centrifuged supernatant poured out and the
deposit is smeared on the clean dry slides and fixed
immediately in ether/95% alcohol for at least 15min and
stained by Papanicolauous method or make the smear, air dry,
and stain with Rowmanowsky stains.

7. 5.ENDOSCOPY SPECIMENS
• Cytological specimens can also be obtained during endoscopy
of the stomach or bronchioles.
• Specimen is centrifuged, supernatant poured out and the
deposit smeared on the clean dry slide and fixed immediately
in ether/ 95% alcohol for at least 15mn and stained by
Papanicolauous method or make smear air dry and stain with
Romanowsky stains.

8. 6.SPUTUM
Procedure
• Add about 15ml of 18% HCl to the specimen and shake to
loosen it from the sides of container.
• Leave it overnight at 20-25˚C. If the specimen can not be dealt
within 24hours use 4% HCl.
• The following morning pour the content which is now liquid
into a plastic universal container, centrifuge,pour off
supernatant, wash by scott tape water, pour off by wire loop
and fix immediately in ether/95% alcohol for at least 15min
and stain by either Papanicolaous method or make smear air
dry and stain with Rowmanowisk stains.

9. 7.OTHER BODY FLUIDS
• It includes
-Pleural fluid(lung)
-Peritoneal (stomach)
-Pericardium(heart)
-ascitic fluid (peritonial cavity)
• Body fluids clots due to protein contents unless anticoagulant is added e.g.
sodium citrate, heparin and EDTA.
• Specimen is centrifuged supernatant poured out and the deposit smeared
on the clean dry slides and fixed immediately in ether/95%alcohol for at
least 15min and stained by Papanicolauous method.
• Or make smear air dry and stain with Rowmanowsky stains.

10. 8.SEMEN
• Is a cytological specimen only obtained from men.
• Semen consists of
-spermatozoa
-seminal fluid(watery part of the semen)
• Examination of semen is called SEMEN ANALYSIS.

11. NOTE.
• For a semen analysis a fresh semen sample is needed after
sexual abstinence for at least 3days.
• The sample can be obtained/collected by different methods,
some of them are
• 1.Masturbation
• 2.Coitus interrupts( withdraw method)
• 3.Prostate massage
The entire sample should be directed into a clean,dry,wide
glass or plastic container.
The patient should bring the specimen to the lab within 1hour.

12. PROCEDURES
MACROSCOPICALLY
1. NOTE THE MACROSCOPIC APPEARANCE
To see whether it is normal(milky/creamy looking)or
abnormal.
2. VOLUME
Measure the volume and record the amount(normal range
is 1.5-5.5ml)
3. VISCOSITY
a)Normal-(string test is 1cm-2cm)
b)Decreased-(stringing less than 1cm)
c)Increased-(stringing greater than 2cm)

13. MICROSCOPICALLY
4.MOTILITY
-Quantitative-estimate % motile vs. immotile
-Qualitative-Grade the forward progression made by the largest
number of spermatozoa.
-The grades are:
-None
-Poor
-Good
-Excellent
Normal motility is greater than or equal to 60% motile
spermatozoa with good to excellent forward progression.
Also we can see other cells like leukocytes ,epithelial cells, Tv and
bacteria.

14. 5.MORPHOLOGY.
Normal morphology-oval head, middle piece
and long-thin tail.
Abnormal morphology of spermatozoa include
-Blunt head
-Multiple tails
-Multiple heads
-Enlarged middle piece
-Very short or very long tail

15. Staining
• The stain techniques used to stain
spermatozoa include Giemsa and
Papanicolaou stains.
• Stain characteristics
Nucleus of head…...................Dark blue
Cytoplasm of head………………Pale blue
Middle piece and tail…….Pink-Red

19. 5.COUNT
• Dilute the semen 1:20 with diluent which is composed of
sodium bicarbonate-formalin solution
• Mix well by taking 1ml of semen and 19mls of the diluent,
leave the mixture to stay for at least 5 minutes,
• Charge the neubauer counting chamber and count at X20
objective.

20. First large square counted
© Oozoa Biomedical Inc, April 2005

21. CONCLUSIONS.
• Good semen according to motility, count and morphology(if
all parameters are good)
• Poor semen according to motility, count and morphology(if all
parameters are not good)
• Good motility, good count but poor morphology, or otherwise
depends on which parameter is good and which is poor.

22. Conclusion cont…
NOTE
 Normal count is 20-60million per ml.
 Less than 20million per ml is-OLIGOSPERMIA.
 No spermatozoa at all-AZOOSPERMIA.

23. THANK YOU FOR LISTENING

24. PREPARED BY
• JOSEPH KITUKULU- RIGHT
• MICHAEL PETER- MIDDLE
• KAUNALA AZIZI-LEFT
LABORATORY SCIENTISTS AT BUGANDO
HOSPITAL, MWANZA TANZANIA
josephkitukulu@yahoo.com