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)
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
1.URINE• Urine assist in the detection of malignancy arising from the urinary system.PROCEDURE1.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 mountedb)-Air dried smear are stained by Rowmanowsky stains e.g. -Giemsa -Field -Leishman
2.CSF• CEREBRAL SPINAL FLUID:Is commonly obtained by lumber puncher.• PROCEDUDE1)Centrifuge immediately after collection2)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
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.
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.
6.SPUTUMProcedure• 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.
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.
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.
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.
PROCEDURES MACROSCOPICALLY1. 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)
MICROSCOPICALLY4.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.
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
Staining• The stain techniques used to stain spermatozoa include Giemsa and Papanicolaou stains.• Stain characteristicsNucleus of head…...................Dark blueCytoplasm of head………………Pale blueMiddle piece and tail…….Pink-Red
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.
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.
Conclusion cont…NOTE Normal count is 20-60million per ml. Less than 20million per ml is-OLIGOSPERMIA. No spermatozoa at all-AZOOSPERMIA.