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Clinical MicroscopyWhat is Clinical Microscopy?- It is an analysis and examination that involves non-blood procedures.

- Involves Urinalysis, fecalysis (stool examination) and Body Fluids.HistoryLaboratory medicine began 6000 years ago with the analysis of human urine, which was called uroscopy until the 17th century and today is termed urinalysis. Today physicians use urine to diagnose selective conditions but from ancient times until the Victorian era, urine was used as the primary diagnostic tool.HistoryUroscopy, from the word 'uroscopia,' means 'scientific examination of urine.' The word is derived from the Greek 'ouron' meaning 'urine' and 'skopeo', meaning to 'behold, contemplate, examine, inspect'.HistorySanskrit medical works from 100 BC describe 20 different types of urine. Hindu cultures were aware that some people's urine tasted sweet, and that black ants were attracted to this sweet urine, a characteristic of the disease now known as diabetes mellitus.

HistoryIn the fourth century BC, Hippocrates (460355 BC) hypothesized that urine was a filtrate of the humors, which came from the blood and was filtered through the kidneys, a fairly accurate description.HistoryGalen (AD 129200) began his scientific findings in Rome. Galen refined Hippocrates' ideas, theorizing that urine represented, not a filtrate of the four humors and overall condition, but rather, a filtrate of the blood. HistoryDuring the renaissance, uroscopy entered the household through the bestselling bookFasiculus Medicinae, published in 1491 by Johannes de Ketham from Germany. De Ketham explained current theories and included a self-diagnostic color wheel, with which individuals could self-diagnose their condition.

HistoryIn 1800, Bichat (17711802), a young pathologist, published a book in which, for the first time, morbid anatomic and histopathologic changes of various organs of the body were discussed and illustrated. Soon thereafter the microscope became an indispensable laboratory tool at medical schools all around the world.

Urinalysis

What is Urinalysis?It is a physical, chemical, and microscopic examination of urine.

- It involves a number of tests to detect and measure various compounds that pass through the urine (Vorvick, 2011)Purpose of Urine testsTo screen for metabolic and kidney disorders and for urinary tract infections.

A urine test can determine whether or not a woman is ovulating or pregnant.

Timed Specimen:

24-hourQuantitative chemical tests, hormone studies

12-hourAddis count

2-hr PostprandialDiabetic monitoring

Afternoon SpecimenUrobilinogen determination

Glucose Tolerance TestAccompaniment to blood samples in GTT

Types of Urine Specimens

RandomRoutine screeningFirst MorningRoutine screeningPregnancy TestsOrthostatic ProteinuriaFasting/Second MorningDiabetic screening / monitoringCatheterizedBacterial cultureMidstream clean-catchRoutine screeningBacterial cultureSuprapubic aspirationBladder urine for bacterial cultureCytologyThree-glass collectionProstatic infectionDrug SpecimenDrug testing3 Stages of UrinalysisA complete urinalysis consists of three distinct testing phases:physical examination, which evaluates the urine's color, clarity, and concentration;chemical examination, which tests chemically for 9 substances that provide valuable information about health and disease; andmicroscopic examination, which identifies and counts the type of cells, casts, crystals, and other components, such as bacteria and mucus, that can be present in urine.

Physical Examination*Color. Many things affect urine color, including fluid balance, diet, medicines, and diseases. How dark or light the color is tells you how much water is in it. Vitamin B supplements can turn urine bright yellow. Some medicines, blackberries, beets, rhubarb, or blood in the urine can turn urine red-brown.

Clarity. Urine is normally clear. Bacteria, blood, sperm, crystals, or mucus can make urine look cloudy.

Odor. Urine does not smell very strong, but has a slightly "nutty" odor. Some diseases cause a change in the odor of urine. For example, an infection with E. coli bacteria can cause a bad odor, while diabetes or starvation can cause a sweet, fruity odor.Chemical Examination*Specific gravity. This checks the amount of substances in the urine. It also shows how well the kidneys balance the amount of water in urine. The higher the specific gravity, the more solid material is in the urine.

pH. The pH is a measure of how acidic or alkaline (basic) the urine is. A urine pH of 4 is strongly acidic, 7 is neutral (neither acidic nor alkaline), and 9 is strongly alkaline. Sometimes the pH of urine is affected by certain treatments. Chemical Examination*Protein. The protein test pad measures the amount ofalbuminin the urine. Normally, there will not be detectable quantities. When urine protein is elevated, a person has a condition calledproteinuria; this can be an early sign ofkidney disease. Albumin is smaller than most other proteins and is typically the first protein that is seen in the urine when kidney dysfunction begins to develop.Chemical Examination*Glucose. Glucose is normally not present in urine. When glucose is present, the condition is called glucosuria. It results from either:An excessively high glucose concentration in the blood, such as may be seen with people who have uncontrolleddiabetes mellitusA reduction in the "renal threshold." Whenblood glucose levelsreach a certain concentration, the kidneys begin to excrete glucose into the urine to decrease blood concentrations. Chemical Examination*Ketones. Ketones are not normally found in the urine. They are intermediate products of fat metabolism. They can form when a person does not eat enough carbohydrates (for example, in cases of starvation or high-protein diets) or when a person's body cannot use carbohydrates properly. Chemical Examination*Blood (Hemoglobin). This test is used to detect hemoglobin in the urine (hemoglobinuria). Hemoglobin is an oxygen-transporting protein found inside red blood cells (RBCs). Its presence in the urine indicates blood in the urine (known ashematuria).

Bilirubin. Bilirubin is not present in the urine of normal, healthy individuals. Bilirubin is a waste product that is produced by the liver from the hemoglobin of RBCs that are removed from circulation. It becomes a component of bile, a fluid that is secreted into the intestines to aid in food digestion.Microscopic ExaminationRed Blood Cells (RBCs). Normally, a few RBCs are present in urine sediment.Inflammation, injury, or disease in the kidneys or elsewhere in the urinary tract, for example, in the bladder orurethra, can cause RBCs to leak out of the blood vessels into the urine. RBCs can also be a contaminant due to an improper sample collection and blood from hemorrhoids or menstruation.Microscopic ExaminationWhite Blood Cells (WBCs). The number of WBCs in urine sediment is normally low. When the number is high, it indicates an infection or inflammation somewhere in the urinary tract. WBCs can also be a contaminant, such as those from vaginal secretions.Microscopic ExaminationEpithelial Cells. Normally in men and women, a few epithelial cells from the bladder (transitional epithelial cells) or from the external urethra (squamous epithelial cells) can be found in the urine sediment. Cells from the kidney (kidney cells) are less common. Microscopic ExaminationMicroorganisms (bacteria, trichomonads, yeast). In health, the urinary tract is sterile; there will be nomicroorganismsseen in the urine sediment. Microorganisms are usually reported as "none," "few," "moderate," or "many" present per high power field (HPF).Microscopic ExaminationTrichomonads areparasitesthat may be found in the urine of women or men (rarely). As with yeast, the trichomonads are actually infecting the vaginal canal and their presence in urine is due to contamination.Microscopic ExaminationCasts. Casts are cylindrical particles sometimes found in urine that are formed from coagulated protein secreted by kidney cells. Under the microscope, they often look like the shape of a "hot dog" and in healthy people they appear nearly clear. This type of cast is called a "hyaline" cast.Microscopic ExaminationCrystals. Urine contains many dissolved substances (solutes) waste chemicals that the body needs to eliminate. These solutes can form crystals, solid forms of a particular substance, in the urine if:The urinepHis increasingly acidic or basic;The concentration of dissolved substances is increased; andThe urine temperature promotes their formation.Fecalysis

What is Fecalysis?Routine fecal examination includes macroscopic, microscopic, and chemical analyses for the detection of gastrointestinal bleeding, liver and biliary duct disorders, malabsorption syndromes, and the detection and identification of pathogenic bacteria and parasites.

Specimen CollectionAdultPassed out in a well rinsed bed panGet a pea size stool from the middle partPlace in a transport vialAvoid mixing the specimen with urine or water from the toiletURINE has harmful effect to protozoans; alter the pH, ammonia, and chloride contentWATER FROM TOILET may contain chemical disinfectantsPreservation of Stool SpecimenPHYSICALFreezingRefrigerationCHEMICAL1 part stool: 3 parts preservativeFormalin40% Ethanol20% glycerin in NSSMerthiolate Iodine FormaldehydePolyvinyl AlcoholPhysical ExaminationAppearanceCauseSoft to Well FormedNormalSmall, hard, dark ballConstipationVoluminous, odorous, floatingMalabsorption of fats or proteinsLoose with mucusVillous adenomaLoose with blood and mucusInflammatory bowel syndromeSticky, black, tarryUpper GIT hemorrhagePhysical ExaminationColor normally light brown to dark brownSubstances responsible for the color of the stoolStercobilinUrobilinHydrobilinPhysical ExaminationpH/ReactionNormal neutralMucusTransluscent gelatinous mucus clinging to the surface of formed stoolBloodIndicates bleedingHEMOFECIA presence of blood in stoolChemical ExaminationOccult Blood hidden bloodScreening test for carcinomaPatient will undergo 3-4 days fish and meat free dietGUIAC TEST routine testGuiac testBlue color indicates gastrointestinal bleeding

NEGATIVEPOSITIVEChemical ExaminationBilirubin Gmelins concentrated HNO3POSITIVE: GREENUrobilinogen Ehrlichs reagentPOSITIVE: CHERRY REDUrobilin Schlesingers testPOSITIVE: GREENISH FLUORESCENCE

Microscopic ExaminationCellsEpithelial cellsPus cellsRBCMacrophage

Food RemnantsBacteriaMucus threads

CrystalsCalcium oxalateTriple phosphateCharcot-leyden

Microscopic ExaminationWhite blood cellsNeutrophils are seen in the feces in conditions that affect the intestinal wall, such as ulcerative colitis and infection with invasive bacterial pathogensParasitesPresence of a parasite or worm-like infection of the intestine, or ova.

Semen Analysis

What is Semen Analysis?Asemenanalysis measures the amount of semen a man produces and determines the number and quality ofspermin the semen sample. A semen analysis is usually one of the first tests done to help determine whether a man has a problem fathering a child (infertility).Tests that may be done during asemen analysisinclude:Volume.This is a measure of how much semen is present in one ejaculation.

Liquefaction time.Semen is a thick gel at the time of ejaculation and normally becomes liquid within 20 minutes after ejaculation. Liquefaction time is a measure of the time it takes for the semen to liquefy.

Sperm count.This is a count of the number of sperm present permilliliter (mL)of semen in one ejaculation.Tests that may be done during asemen analysisinclude:Sperm morphology.This is a measure of the percentage of sperm that have a normal shape.

Sperm motility.This is a measure of the percentage of sperm that can move forward normally. The number of sperm that show normal forward movement in a certain amount of semen can also be measured (motile density).

Tests that may be done during asemen analysisinclude:pH.This is a measure of the acidity (lowpH) or alkalinity (high pH) of the semen.

Whitebloodcell count.White blood cells are not normally present in semen.

Fructose level.This is a measure of the amount of a sugar called fructose in the semen. The fructose provides energy for the sperm.Equipment

Urine Strips

Equipment

Automatic Urine AnalyzerEquipmentMicroscope

EquipmentMicroscope slide(s)

EquipmentUrine Sample Container

EquipmentStool specimen container

Referenceshttp://www.annclinlabsci.org/content/32/3/309.fullhttp://pioneerlab.ph/Clinical_Microscopy_(P).htmlhttp://www.nature.com/ki/journal/v71/n5/full/5002057a.htmlhttp://www.bd.com/vacutainer/labnotes/Volume14Number2/http://labtestsonline.org/understanding/analytes/urinalysis/ui-exams/start/2http://labtestsonline.org/understanding/analytes/urinalysis/ui-exams/start/1http://www.webmd.com/infertility-and-reproduction/guide/semen-analysis

Thank you!Presented by Group 3:David, FlorielleGamboa, Lois DanielleLacson, Maria MichaelaLusung, Ashley JaneQuiambao, Mark RaysonSotto, Kristel JoyVelayo, Erika