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This document summarizes key information about seminal fluid and synovial fluid. It describes the composition and functions of seminal fluid and the processes of sperm production and ejaculation. It also outlines normal ranges for semen analysis tests. For synovial fluid, it discusses the collection and examination of samples, including appearance, cell counts, and testing for crystals. Both fluids are complex body fluids that are analyzed to evaluate health and diagnose conditions.
This document summarizes key information about seminal fluid and synovial fluid. It describes the composition and functions of seminal fluid and the processes of sperm production and ejaculation. It also outlines normal ranges for semen analysis tests. For synovial fluid, it discusses the collection and examination of samples, including appearance, cell counts, and testing for crystals. Both fluids are complex body fluids that are analyzed to evaluate health and diagnose conditions.
This document summarizes key information about seminal fluid and synovial fluid. It describes the composition and functions of seminal fluid and the processes of sperm production and ejaculation. It also outlines normal ranges for semen analysis tests. For synovial fluid, it discusses the collection and examination of samples, including appearance, cell counts, and testing for crystals. Both fluids are complex body fluids that are analyzed to evaluate health and diagnose conditions.
Seminal fluid or semen complex body fluid used to transport sperm or spermatozoa - analyzed routinely to evaluate fertility and to follow up after a vasectomy to ensure its effectiveness - evaluation of quality for donation purposes and forensic application - composed primarily of secretions from the testes, epididymis, seminal vesicles, and prostate gland, with a small amount derived from bulbourethral glands - sperm production is regulated by Sertoli cells (in seminiferous tubules) which functions as a barrier
Testes exocrine function (secretion of sperm) - endocrine function (secretion of testosterone)
Epididymis tubular network wherein the luminal fluid from sertoli cells are carried for the maturation of the immotile and immature sperm - serves in the concentration of the sperm by absorption of lumen fluid and their storage until ejaculation
Seminal vesicles and prostate gland serves as accessory gland of the male reproductive system - produce and store fluids that provide the principal transport medium for sperm
Specimen collection It should be collected in a plastic or glass container , or in a special type of condom known as a collection condom or through masturbation. It should be performed after a 48- 72hr absence of sexual activity so it contains an accurate account of sperm count and viability. The specimen should arrive at the laboratory as soon after collection as possible so that an accurate Liquefaction time can be reported. During transportation it should be kept near body temperature. The patient should be asked if any of the sample is lost since the highest concentration of sperm is in the first part of ejaculation.
MFG*ILMT MLS IV-B
Apperance of semen Normal semen: grayish white and opalescent If brown or red presence of blood If yellow associated with certain drugs If turbid with less translucent there is an increase in large numbers of leukocytes Odor: musty
Volume of semen Analysis of seminal fluid should take place immediately following liquefaction not more than 60 minutes after collection. A normal, complete ejaculate collection recovers 2 to 5mL of seminal fluid. Viscosity of semen After complete liquefaction, the viscosity of the semen is evaluated using Pasteur pipette and observing the droplets that form when the fluid is allowed to fall by gravity A normal specimen is watery and forms into discrete droplets. Abnormal viscosity or fluid thickness is indicated by the formation of a string or thread greater than 2cm length. Motility of semen Laboratory evaluation of sperm motility is performed by examining the undiluted specimen microscopically and determining the percentage of sperm showing active motility. Approximately 25 high-power fields should be examined. - A minimum motility of 50-60 percent with a quality of fair (2.0) is considered normal for specimens tested within the 3-hour time period. Motility Grading Criteria 0 immotile 1 motile, without forward progression 2 motile, with slow nonlinear or meandering progression 3 motile, with moderate linear (forward) progression 4 motile, with strong linear (forward) progression
Concentration and sperm count Normal concentration: 20-250 million per mL of sperm - determined by using a hemacytometer after preparing an appropriate dilution of 1:20 Sperm count= sperm concentratio n(sperm/mL) x Volume of ejaculate (mL)
Morphology Normal sperm: oval head measuring 3x5 m and a long tapering tail Abnormality: associated with poor ovum penetration include double heads, giant and morphous heads, pin heads, tapering heads, and constricted heads MFG*ILMT MLS IV-B
double or coiled tails impeded motility immature sperm (spermatids) may be present and must be differentiated from WBC
**Sperm morphology should be reported from a stained specimen examined under oil immersion, the recommended stain is the hematoxylin, crystal violet, or Giemsa stains. **Additional tests: sperm viability, seminal fluid fructose level and sperm agglutinins
Normal semen analysis Semen volume: 2ml or more Semen pH: 7.2-8.0 Liquefaction time: 20-30 mins after collection Sperm collection: 20 million spermatozoa per ejaculate or more Sperm count: 40 million spermatozoa per ejaculate or more Sperm morphology: more than 30% are normal Sperm motility: 50% show progressive movement; 25% or more with rapid progressive movement Sperm vitality: 75% or more live WBC: fewer than 1 million WBC/ml
Semen Biochemistry Acid phosphatase: marker for prostatic function Citric acid: can indicate prostatic function low levels may indicate dysfunction or a prostatic duct obstruction Zinc: marker for prostatic function colorimetric assay (WHO) Fructose: marker for seminal vesicle function, and is a substrate for sperm metabolism spectrophotometric assay (WHO) -Glucosidase: secreted exclusively by the epididymis and so is a marker for epididymal function spectrophotometric assay (WHO
SYNOVIAL FLUID
Synovial fluid - normally thick, straw-colored liquid in small amounts in joints, bursae (fluid-filled sacs in bones) and tendon sheaths - lubricates the joint and allows for ease movement - ultrafiltrate or dialysate of plasma and contains levels of glucose and uric acid
Specimen collection: After the joint area is cleaned, the health care provider inserts a sterile needle through the skin and into the joint space. Fluid is then drawn through the needle into a sterile syringe. The fluid sample is sent to the laboratory. The laboratory technichian: MFG*ILMT MLS IV-B
o checks the sample's color and clarity o places the sample under a microscope, counts the number of red and white blood cells, and looks for crystals (in the case of gout) or bacteria o measures glucose, proteins, uric acid, and lactic dehydrogenase (LDH) o cultures the fluid to see if any bacteria grow Normal synovial fluid: will not clot Abnormal synovial fluid: fluid from diseased joint may contain fibrinogen and form a clot **therefore, both anticoagulated and non-anticoagulated specimens should be collected
Recommended specimen containers: 1. EDTA tube (Lavender top tube) for cell counts, differentials and viscosity. 2. Heparanized tube (dark green top tube) for chemistry and immunologic III-47 tests 3. Plain sterile tube or syringe (with needle removed) for microbiologic testing and crystal examination
Appearance Color: should be evaluated in a clear glass tube against a white background colorless but is often pale yellow because of diapedesis of a few RBCs associated with even mild trauma straw to yellow colored (xanthochromia) seen in noninflammatory and inflammatory disorders
Clarity: relates to the number and types of particles within the synovia normal synovial fluid: transparent; newsprint is easily read through translucent fluid: obscures details but black and white areas can be distinguished opaque fluid: completely obscures background leukocytes are most commonly responsible for changes in clarity, however, very large numbers of crystals may produce an opaque, milky fluid without leukocytes shimmering, oil-appearing specimen abundance of cholesterol crystals which may grossly resemble pus increased turbidity less often due to concentration of fibrin, free-floating rice bodies, metal and plastic particles from patients with joint prostheses, or cartilage fragments in osteoarthritis
Viscosity: normal synovial fluid: viscid and does not clot difference from other fluids derived from plasma: high content of hyaluronic acid (mucin)
Total cell counts total leukocyte counts should be performed promptly o degenerative loss begins as soon as one hour following arthrocentesis cell counts are usually performed in a standard hemocytometer MFG*ILMT MLS IV-B
leukocyte counts: > 10,000/UI o > 50,000/UI crystal induces arthritis chronic inflammatory arthritis septic arthritis o < 10,000/UI osteoarthritis osteochondritis dissicans trauma synovioma upper reference level for leukocytes: 150-200/uL
Recommended tests Routine Tests Gross examination (color,clarity) Total and differential leukocyte counts Gram stain and bacterial culture (aerobic and anaerobic) Crystal examination with polarizing microscope and compensator Crystal identification Types of Crystals Gout and pseudogout most common crystalline arthropathies - caused by deposition of monosodium urate (MSU) and calcium pyrophosphate dehydrate (CPPD) crystals normal joint fluid is negative for any crystals
- maybe seen following intraarticular injections - iatrogenic - strongly variably birefringent - polymorphic clumps, rods and rhomboids - may cause joint pain for several hours
Cholesterol crystals - may be present in chronic effusions from patients with osteoarthritis or RS - very large - flat and rectangular shaped - notched corners - varying birefringence - rheumatoid arthritis
Calcium phosphate (apatite) crystals - small chunky rods - seen in calcific periarthritis, osteoarthritis and inflammatory arthritis
Slide preparation A small amount of hyaluronidase should be added to synovial fluid prior to performing cell counts or preparing cytocentrifuge slides to liquefy the fluid
Mucin Clot Test - Ropes Test - estimation of the integrity of the hyaluronic acid-protein complex (mucin) - normal fluid forms a tight ropy clot upon the addition of acetic acid - good mucin clot: indicates good integrity of the hyaluronate - poor mucin clot: one that breaks up easily; associated with destruction or dilution of hyaluronate
Chemical Tests Glucosetypically a bit lower than blood glucose levels; may be significantly lower with joint inflammation and infection Proteinincreased with bacterial infection Lactate dehydrogenaseincreased LD (LDH) level may be seen in rheumatoid arthritis, infectious arthritis, or gout
Microbiologic Tests Normal synovial fluid: has small numbers of WBCs and RBCs but no microorganisms or crystals present Culture and susceptibility testing determine what type of microorganisms present
Infectious Disease Test MFG*ILMT MLS IV-B
- in addition to chemistry tests, other tests may be performed to look for microorganisms if infection is suspected
Serologic Tests Rheumatoid Factor (RF) antibody to immunoglobulins - present in the serum of patients with RS - false positive RF can result from other chronic inflammatory diseases
AMNIOTIC FLUID
Amniotic fluid found around the developing fetus, inside a membranous sac, called amnion - formed in the placenta - volume increases from: o 30ml at 10 weeks, o 450ml at 20 weeks o up to 800-1000 ml at 37 weeks - serves as cushion, protection and serves as the key role in the exchange of water and molecules between the fetus and the maternal circulation - the fluid absorbs jolts, prevent adherence of the embryo to the amnion and allows fetal movement
Hydramnios- increase in amniotic fluid volume (1,500-2,000ml) - caused by maternal diabetes, congenital malformations and gastrointestinal defects that prevents FETAL SWALLOWING.
Oligohydramnios- decreased amount of amniotic fluid (less than 400ml) - occur with premature rupture of the membranes and with congenital malformations.
The amniotic fluid uses to assess the status of the fetus Tests to diagnose genetic and congenital disorders before birth Test to detect fetal distress from HEMOLYTIC DISEASE OF THE NEWBORN or from infection Test to assess FETAL LUNG MATURITY To assess the ability of the fetus to survive early delivery
Amniocentesis obtained by needle aspiration 15-18 weeks of gestation for genetic studies 10-20ml (maximum 30ml) dispensed into sterile plastic specimen containers cell culture and chromosomal studies stored at body or room temp MFG*ILMT MLS IV-B
phospholipid analysis transported on ice and centrifuged at 500g
Appearance Normal amniotic fluid: colorless to pale yellow Slightly turbid due to fetal cells, vernix and hair Dark yellow or amber color associated with bilirubin Green color indicates meconium; newborn first fecal bowel movements Very dark red brown associated with fetal death
Difference of amniotic fluid to maternal urine Urine: higher levels of creatinine and urea Amniotic fluid: higher levels of glucose and protein
Testing for neural tube defects- alpha fetoprotein and acetylcholine esterase anencephaly spina bifida high levels of AFP AChE is more specific than AFP testing
Fetal distress testing 1. HDN or Erythroblastosis Fetalis when a mother develops an antibody to an antigen of fetal erythrocytes 2. Infection vaginal vaginosis and trichomoniasis have been linked to preterm delivery and spontaneous abortion 3. Respiratory distress syndrome most common death in premature and newborn immature fetal lungs, lack of surfactant (allows alveoli to function normally during inhalation and exhalation)
Fetal lung maturity test 1. Lecithin: Sphingomyelin Ration and Phosphatidylglycerol - levels of lecithin and sphingomyelin are relatively equal - after 34 weeks, levels of sphingomyelin decreases, while levels of lecithin increases - (L/S) ration of 2.0 or greater is associated with fetal pulmonary system maturity - Phosphatidylglycerol detectable only 35 week of gestation (not affected by blood and meconium) 2. AMNIOSTAT-FLM - uses antibodies against phosphatidyl glycerol to detect fetal lung surfactant. 3. FOAM STABILITY - screening test 4. MICROVISCOSITY FLOURESCENCE POLARIZATION ASSAY - this assay provides flourescence polarization surfactant: albumin ratio 5. LAMELLAR BODIES - provide reliable estimate of fetal lung maturity
Blood-brain barrier - tight junction between capillary endothelial cells and epithelial cells in the choroid - prevent some substances to enter CSF - small molecules and lipid soluble substances pass through easily
Function: cushion of the brain and spinal column serves as a nutrient and metabolic waste exchange fluid adjust its volume in response to changes in cerebral vessel changes
Specimen collection Lumbar puncture: Adults intervertebral space between L3 and L4 Small children and infants intervertebral space between L4 and L5
Physical characteristics clear colorless viscosity similar to water
Traumatic Tap Blood distribution: there is significant difference in the amount of blood present between first and last tube Clot formation: may form clot with bloody fluid
Hemorrhage Blood distribution: homogenous amount of blood present in all tubes Clot formation: may form clot but with the absence of bloody fluid
Cell Count Specimen used: well-mixed, undiluted specimen Counted in Neubauer counting chamber Cells normally found o PMNs o Monocytes MFG*ILMT MLS IV-B
Lymphocytic pleocytosis Later stage of viral, tubercular, fungal, syphilitic meningitis Increased number of leukocytes in Guillian-Barr syndrome
Plasmacytes Multiple sclerosis
Eosinophils Parasitic and fungal infections Allergic reactions
Mixed pleocytosis Chronic bacterial meningitis Meningitis of fungal or tuberculin origin Rupture of cerebral abscess
Other cells in normal CSF: Ependymal cells Choroidal cells PAM cells MFG*ILMT MLS IV-B
Chemical analysis 1. Protein - Total protein: 15-45 mg/dL - comprised by low molecular protein Methods: dye-binding immunochemistry modified biuret methods turbidimetric methods - increased in endocrine disorders, traumatic tap and infections - decreased in hyperthyroidism o Albumin derived from transport across the blood-brain barrier o Protein electrophoresis presence of oligoclonal bands may establish diagnosis 2. Glucose - 60-70% of plasma in normal adults - normal range: 50-80 mg/dL - increased in hyperglycemia and traumatic tap - decreased in CNS infections, hypoglycemia, impaired glucose transport and metastatic carcinoma 3. CSF Lactate - present in CSF due to CNS anaerobic metablosim - levels are independent from plasma lactate levels - used to differentiate viral meningitis (30mg/dL) from other forms of meningitis (>35 mg/dL) - increased levels usually reflect CNS tissue hypoxia 4. CSF Glutamine - produced from ammonia and -ketoglutarate by the brain cells - serves to remove the toxic metabolic waste product ammonia from CNS - elevated levels are associated with liver disorders (Reye Syndrome)
Microbiology procedures meningitis is the most serious diagnosis the most sterile tube collected is used
Sediment of centrifuged CSF is inoculated into thioglycolate broth and plates of blood agar, chocolate, and MacConkey agar If Haemophilus is suspected, Strips of X-V may be applied to the blood agar plate If fungal meningitis is suspected, Sabouraud dextrose agar should be inoculated If Mycobacteria is suspected, inoculate Middlebrook broth and agar NOTE: It is important to note that if antibiotic therapy was administered prior to the collection of CSF for culture, the recovery of microorganisms may be significantly reduced.
SEROUS FLUID Serous fluid fluid between the membranes which provides lubrication as the surfaces move against each other
Formed under the influence of Hydrostatic pressure Osmotic pressure Capillary permeability
Pleural Fluid - fluid in the lung cavity - it is about 3- 20 ml under normal conditions. - drained by the lymphatic system - normally its a clear or pale yellow fluid. Turbidity: white blood cells and microorganisms Blood: traumatic injury, malignancy, and traumatic tap Milky: chylous or pseudochylous material Neutrophils: bacterial infection Lymphocytes: tuberculosis and malignancy Low glucose: tuberculosis, rheumatoid inflammation and malignancy Low pH: tuberculosis, malignancy and esophageal rupture Elevated amylase: pancreatitis
Pleural Effusion - an abnormal amount of fluid around the lung. - most pleural effusions are not serious by themselves, but some require treatment to avoid problems - causes: o Congestive heart failure o Pneumonia MFG*ILMT MLS IV-B
o Liver disease (cirrhosis) o End-stage renal disease o Nephrotic syndrome o Cancer o Pulmonary embolism o Lupus and other autoimmune conditions - symptoms: o Shortness of breath o Chest pain, especially on breathing in deeply (pleurisy, or pleuritic pain) o Fever o Cough - diagnosis: o Chest X-ray film o Computed tomography (CT scan) o Ultrasound Pericardial Fluid - fluid surrounding the heart - clear and pale yellow - volume 10- 15 ml Milky: lymphatic drainage Tirbidity: infection and malignancy Blood: tuberculosis, tumor and cardiac puncture Neutrophils: bacterial endocarditis Low glucose: bacterial infection and malignancy
Pericardial Effusion - the presence of excessive pericardial fluid within the potential space of pericardium. - rapid accumulation of pericardial fluid may cause elevated intrapericardial pressures with as little as 80 mL of fluid, while slowly progressing effusions can grow to 2 L without symptoms
Cardiac tamponade - occurs when the heart is squeezed by fluid that collects inside the sac that surrounds it - three principal features o Elevation of intracardiac pressures o Limitation of ventricular filling o Reduction of cardiac output - causes: o any condition lead to pericarditis can lead to pericardial effusion . o the most common cause are: Neoplastic disease MFG*ILMT MLS IV-B
Idiopathic pericarditis Uremia Following cardiac operation Trauma - signs and symptoms: o shortness of breath o weakness and fatigue o anxiety o tachycardia o jugular vein engorged o cyanosis - diagnosis: o physical examination o electrocardiogram (ECG) o chest X-ray film
Peritoneal Fluid - fluid in the abdominal cavity - Clear or pale yellow - > 50 ml Turbidity: peritonitis and cirrhosis Blood: trauma Neutrophils: peritonitis Low glucose: tubercular peritonitis and malignancy Elevated amylase: pancreatitis, gastrointestinal perforation Elevated alkaline phosphatase: intestinal perforation Elevated urea or creatinine: ruptures bladder
Peritoneal Effusion - an accumulation of fluid in the peritoneal cavity and is also known as ascites. - other name: hydroperitoneum and abdominal dropsy
Transudate and exudate Transudate - produced as a result of disruption of fluid production and regulation between the serous membranes Exudate - caused by conditions producing damage to the serous membranes
MFG*ILMT MLS IV-B
Ascites Common causes: Transudate 1. Cirrhosis 2. Congestive heart failure 3. Hypoalbuminemia Exudate Metastatic ovarian cancer and infective peritonitis
Signs and symptoms: abdomen related everted umbilicus flank fullness lank dullness( if absent this means that there is < 10% chance of having Ascites) there is at least 1.5 liters of Ascites if dullness is present], shifting dullness fluid thrill
Diagnosis: USG : confirm the diagnosis of minimal amount of ascites Paracentesis
General Laboratory Procedures Routine fluid examination including: - classification as a transudate or exudate - Appearance - cell count (differential) o cell counts are usually performed manually using the Neubauer counting chamber o differential counts are performed on (Wright stained smears) o any suspicious cells been on the differential should be referred to the cytology laboratory or the pathologist - chemistry - microbiology procedures