AUBF Flashcards
CSF is first recognized by ____ in ______
Catugno, 1764
Functions of CSF
Supply nutrient to nervous tissue
Remove metabolic waste
Cushion the brain & spinal cord against trauma
Normal adult volume of CSF
140-170
Normal volume of CSF neonates
10-60
Reduce the amount of CSF in the brain by draining it to the abdomen or directly into the chambers of the heart
Ventricular Shunt
helps reduce the pressure on the brain
Shunting of CSF
Hollow plastic tube that is placed in the ventricle of the brain
Shunt
Ventricular peritoneal shunt located from the
brain to abdomen
Ventricular atrial shunt is located from brain to _____
heart
Lines the brain and the spinal cord
Protective layer
Meninges
directly in contact with skull bones
outermost
dura mater
Web-like structure - middle layer of meninges
Arachnoid
space where CSF flows
Subarachnoid space
directly in contact with brain
Innermost
Pia Mater
Capillary networks that form the CSF
Choroid Plexus
light-fitting endothelial cells: separates blood tissues and brain tissue
Blood Brain Barrier
functional barrier; separates blood tissues and cerebrospinal fluid
Blood CSF Barrier
Normal volume production of CSF
20/mL per hour
Cells that help production of CSF
Ependymal cells
Specimen collection for CSF procedure:
Lumbar tap/Lumbar puncture between 3rd and 4th or 4th and 5th lumbar vertebrae
Opening pressure is measured _____ in aspiration of CSF
First
CSF collection tubes order of draw:
Tube 1: Chemistry or serology
Tube 2: Microbio
Tube 3: Hematology
*Tube 4 if possible: micro
Storage for CSF tube 1
Frozen (0 to frozen)
Storage for CSF tube 3
Refrigerated
Increase in blood CSF may indicate
traumatic tap
Storage for CSF tube 2
Room temperature (20 to 24 deg C)
Appearance of CSF: Cloudy, Milky, Turbid
Protein, Lipid, WBC
Appearance of CSF: Normal
Crystal Clear
Appearance of CSF: Bloody
Intracranial hemorrhage, Traumatic tap
Appearance of CSF: Xanthochromic
Hemoglobin, Bilirubin, Carotene, ↑Protein, Melanin
Appearance of CSF: Pink
Oxyhemoglobin
Appearance of CSF: Yellow
High bilirubin content
Appearance of CSF: Orange
combination of oxyhemoglobin and bilirubin
Appearance of CSF: Clotted
Clotting factors introduced by traumatic tap
Appearance of CSF: Pellicle
Tubercular meningitis, seen after overnight of refrigeration
Appearance of CSF: Oily
Radiographic Contrast Media
Traumatic tap indication:
Uneven blood distribution
Clot formation
Supernatant: Not Xanthochromic
Intracranial Hemorrhage indication:
Even distribution of blood
No clot formation
Supernatant: Xanthochromic
Additional: Erythrophagocytosis: microscopically- macrophages with ingested RBCs
(+): D-Dimer
Cell count that is routinely performed on CSF
Leukocyte/wbc count
When is Leukocyte/WBC count performed on CSF
Traumatic tap
_______ are usually determined only when a traumatic tap has occurred and a correction for leukocytes or protein is desired
RBC counts
RBC disintegrates within
1 hour
CSF: ↓ by 40% in WBC disintegrates after
2 hours
increase in the number of cells in CSF
Pleocytosis
Normal values of WBC in ADULT CSF
0-5 cells/uL
Predominant cells in ADULT CSF
lympthocytes
Predominant cells in neotates CSF
monocytes
Normal values of WBC in neonates
0-30 cells/uL
Manual method for counting cells in CSF
Neubauer Counting Chamber
Counting calls of CSF
4 larger corner squares and large center squares
Area of large square neubauer chamber:
1 mm^2
Total Cell Count:
Cell counted x Dilution / Area x Depth
Needed for the correction of WBC Count & CSF Protein Results
RBC Count
Diluent for CSF RBC count
NSS
Diluent for WBC count CSF:
3% Glacial Acetic Acid
Can be added to diluting fluid of WBC count
Methylene Blue
Correction for Contamination of CSF
WBC added = WBC Blood x RBC CSF
RBC Blood
Subtract 1 WBC for every ____ RBCs present in the CSF
700
CSF Electrophoresis primary purpose:
detection of OLIGOCLONAL bands
Can determine if a fluid is indeed CSF
CSF Electrophoresis
Most Common Method of CSF electrophoresis
Agarose Gel Electrophoresis
Indicates immunoglobulin production
Oligoclonal Bands
Oligoclonal bands are located in the ____ region of the protein electrophoresis
GAMMA
Oligoclonal bands in CSF only
Multiple Sclerosis; Neurologic Disorder, Encephalitis, Neurosyphilis, Guillain Barre Syndrome
Oligoclonal Bands in CSF and Serum
HIV infection
Oligoclonal bands in Serum only; CSF banding because of BBB leakage or blood contamination during lumbar tap.
Leukemia, Lymphoma, Viral Infections
Indicates recent destruction of the myelin sheath that protects the axon of the neurons
Myelin Basic Protein
Myelin Basic Protein can be used to monitor ________-
Multiple sclerosis
Myelin Basic Protein theorized that the inflammatory response in multiple sclerosis is triggered by ________
molecular mimicry
Normal volume of CSF Glucose:
60-70% of plasma glucose
Blood glucose should be drawn ________ prior to lumbar tap (CSF)
2 hours
Can be used to determine the cause of meningitis
CSF glucose
Increased levels of CSF glucose is always a result of
plasma elevations
Decreased levels of CSF glucose
caused by alteration in transport in BBB ↑ used by braincells
Destruction of tissue within the CNS due to hypoxia causes the production of
Increased CSF lactate
Aid in the diagnosis and management of meningitis cases
CSF Lactate
Elevation of CSF lactate is consistent with:
Bacterial, Tubercular, and Fungal Meningitis
Level of CSF lactate when treatment is successful
declines rapidly
Used to monitor severe head injuries
CSF Lactate
Falsely elevated CSF lactate is due to
xanthochromic or hemolyzed samples
Preferred over direct measurement of CSF ammonia
CSF Glutamine
CSF Glutamine normal values
8-18 mg/dL
Elevated levels of CSF glutamine is associated with
liver disorder that result in blood and CSF ammonia
CSF glutamine is produced from ____ and ____ by the brain cell
Ammonia & α-ketoglutarate
______ is an indirect test for the presence of excess ammonia in CSF
Glutamine
Frequently requested procedure for patients with coma of unknown origin
CSF Glutamine
CSF glutamine is ↑ in approximately 75% of children with
Reye Syndrome
Bacterial meningitis is recovered after ____
24 hours
Tubercular meningitis is recovered after ___
6 weeks
CSF centrifuged at _____ for ___ minutes
1500 g for 15 minutes
used for slides and culture of CSF
sediments
Confirmatory rather than Diagnostic Procedure
CSF Culture
Microbiology laboratory test for a preliminary diagnosis of CSF
Gram Stain
Acid Fast Stain
India Ink Preparation
Latex Agglutination Test
Predominant cell in bacterial meningitis
Neutrophil
Predominant cell in Viral meningitis
Lymphocyte
Predominant cell in Tubercular
Lymphocyte, Monocyte
Predominant cell in Fungal meningitis (LEM)
Lymphocytes, Monocytes, Eosinophil
↑ Protein: Marked
Glucose: Markedly ↓
Bacterial meningitis
Increased protein: Moderate
Glucose: Normal
Viral meningitis
↑ Protein: moderate to marked
Glucose: decreased
Tubercular meningitis
↑ Protein: moderate to marked
Glucose: normal to ↓
Fungal meningitis
Lactate: > 35 mg/dL
Bacterial meningitis
Lactate: normal
Viral meningitis
Lactate: >25 mg/dL
Tubercular or Fungal meningitis
Routinely performed on CSF from all suspected cases of meningitis
Gram stain
CSF specimen uses _______ that provides highly concentrated specimens
Cytocentrifuge
Common causes of bacterial meningitis for infants
S.agalactiae
E. Coli K1 strain,
L. monocytogenes
Common causes of bacterial meningitis for adolescent/adults
N. meningitides
H. Influenza type B,
S. pneumoniae
Common causes of bacterial meningitis for elderly
S. pneumoniae
N. Meningitides
L. monocytogenes
one the frequently occuring complications of AIDS
Cryptococcal meningitis
Cryptococcus neoformans india ink result:
Thick encapsulated organism
Cryptococcus neoformans gram stain result:
Starburst pattern (may be seen more often than a positive india ink)
Cryptococcus neoformans is associated with increased _______
eosinophils
Diagnosis of meningitis caused by Gram negative bacteria
Limulus lysate test
reacts with bacterial endotoxin of Gram (-) bacteria
Limulus amebocyte
Reagent for limulus lysate test
blood cells of horseshoe crab (limulus polyphemus)
(+) Result of limulus lysate test
Coagulation within 1 hour of incubation at 37 deg C
Rapid Test for CSF:
Latex
ELISA method
Bacterial Antigen Test
Performed to detect the presence of neurosyphilis
Serologic Testing
Procedure recommended by the CDC to diagnose Neurosyphillis
Venereal Disease Research Laboratories (VDRL)
More sensitive than VDRL
Fluorescent Treponemal Antibody-Absroption (FTA-ABS) Test
Prevent contamination with blood because the FTA-ABS remains —— in the serum of treated cases of syphilis (Treponema pallidum)
positive
Semen - composed of four fractions that are contributed by the: (TSPB)
Testes (Epididymis)
Seminal vesicles
Prostate gland
Bulbourethral gland
paired glands in the scrotum that contain the seminiferous tubules
Testes
production of spermatozoa takes place
seminiferous tubules
storage of sperm
epididymis
Seminal Vesicles produces most fluid present in the semen (percentage)
60-70%
Seminal fluid contains high concentration of
fructose
use by sperm for energy needed for motility
fructose
responsible for gray appearance color of the sperm
flavin
aids in propelling the sperm through the urethra by contractions during ejaculation
Prostate Gland
percentage of acidic fluid produced by the prostate gland
20-30%
Milky acidic fluid from prostate gland contains:
It is also responsible for both coagulation and liquefaction of semen following ejaculation
Acid phosphatase
Citric acid
Zinc
Proteolytic enzymes
Bulbourethral gland contributes about ___ of the fluid volume
5%
Sexual abstinence for semen analysis:
at least 2 days but nor more than 7 days
Prolong abstinence
Higher volume and decreased motility
Fertility testing according to WHO:
2 or 3 sample be collected not less than ___ or more than ___
7 days or more than 3 weeks
Abnormal semen analysis result according to WHO
2 abnormal samples are considered significant
Specimen for semen analysis should be kept at _____ temp and delivered within ____
room temperature
(20-24) within 1 hour
Specimen awaiting for semen analysis should be kept
37 deg C
Container needed for semen analysis
only non-lubricant-containing rubber or polyurethane condoms should be used
Ordinary condom contains:
spermicides
Reference value for semen analysis: Volume
2-5 mL
Reference value for semen analysis: Viscosity
pours in droplets
Reference value for semen analysis: pH
7.2 - 8.0
Reference value for semen analysis: Sperm concentration
> 20 million/mL
Reference value for semen analysis: Sperm count
> 40 million/ejaculate
Reference value for semen analysis: motility
> 50% within 1 hour
Reference value for semen analysis: quality
> 2.0 or a,b,c
Reference value for semen analysis: morphology (strict criteria)
> 14% normal forms
Reference value for semen analysis: morphology routine
> 30% normal forms
Reference value for semen analysis: round cells
<1.0 million/mL
Sperm Motility Grading: 4.0
Rapid, straight-line motility
Sperm Motility Grading: 3.0
Slower speed, some lateral movement
Sperm Motility Grading: 2.0
Slow forward progression, noticeable lateral movement
Sperm Motility Grading: 1.0
no forward progression
Sperm Motility Grading: 0
no movement
Alternative sperm motility grading criteria:
Progressive Motility
Sperm moving linearly or in a large circle
Alternative sperm motility grading criteria:
Non-progressive (NP)
Sperm moving with an absence of progression
Alternative sperm motility grading criteria:
Immotility
No movement
Normal semen appearance:
gray-white color, appears translucent, has characteristic of musty odor/bleach like odor; opacity (due to flavin)
Semen appearing almost clear
Low sperm concentration
Increase turbidity of semen indicates
presence of WBCs and infection within the reproductive tract
Useful in differentiating presence of WBC compared to immature sperm (spermatids)
Leukocyte esterase
Leukocyte esterase test positive result indicates
WBC
Leukocyte Esterase negative result
spermatids
Red coloration of semen is due to
presence of RBC
Yellow coloration of semen
urine contamination (toxic to sperm), prolonged abstinence, and medication
Fresh semen specimen is clotted and liquefy within ______ after collection
30 to 60 minutes
Failure of liquefaction within 60 minutes: caused by deficiency in
prostatic enzymes
If after 2 hours the specimen has not liquefied: an equal volume of physiologic ______ may be added to induce liquefaction and allow the rest of the analysis to be performed
Dulbecco’s phosphate-buffered saline or proteolytic enzymes (alphachymotrypsin or bromelain)
Normal semen volume
2-5 mL
Increased semen volume is due to
prolonged abstinence
Decreased volume of semen is associated with
infertility
incomplete specimen collection must be considered
Refers to the consistency of the fluid and may be related to specimen liquefaction
Viscosity
Viscosity: Incomplete liquefied specimen
clumped and highly viscous
Normal viscosity of semen:
easily drawn into a pipette and form small discrete droplets, do not appear clumped or stingy
Droplets that form longer than 2cm of semen
highly viscous, abnormal (problem with enzyme)
Normal pH of semen
7.2 to 8.0 (alkaline)
Increased pH of semen indicates:
infection within the reproductive tract
Decreased pH of semen:
increased prostatic fluid, ejaculatory duct obstruction, poorly developed seminal vesicles
Normal Sperm Concentration
greater than 20- 250 million sperm per milliliter
Borderline Sperm Concentration
10-20 million sperm per milliliter
Computation for sperm count:
Sperm count: Sperm concentration x Specimen Volume
Dilution of semen to diluting fluid:
Dilution: 1:20
Sperm should be capable of ______, progressive motility
forward
Motility should be assessed within
1 hour of collection
Head of sperm morphology:
oval shape approximately 5 um long and 3um wide
enzyme containing critical to ovum penetration;
(encompasses half of the head and cover approximately two-thirds of the sperm nucleus)
acrosomal cap
Enzyme present in acrosomal cap:
Hyaluronidase & Acrosin
attaches the head to the tail and midpiece
neckpiece
approximately 7 um long, thickest part of the tail (surrounded by mitochondrial sheath that produces energy required by tail for motility)
Midpiece
inherited only from the mother. Used for patient identification.
mtDNA
Tail - approximately ___ long
45 um
Problem with sperm tail
decreased motility
Problem with sperm head
ovum penetration
stain for sperm vitality by counting the number of dead cells in 100 sperm using bright field or phase contrast microscopy
Eosin nigrosin
Normal sperm concentration with markedly decreased motility
decreased sperm vitality
remains bluish white in eosin nigrosin stain for sperm
living cells
stain red against purple background in eosin nigrosin stain for sperm
dead cells
Normal Vitality for sperm
50% or more living cells
Normal seminal fluid fructose
≥13 umol per ejaculate
Low seminal fluid fructose indicates:
abnormalities of the seminal vesicle
bilateral congenital absence of the vas deferens
obstruction of the ejaculatory duct
partial retrograde ejaculation and androgen deficiency
Seminal fluid fructose should be tested within ___ or ___ to prevent fructolysis
2 hours or frozen to prevent fructolysis
present in both men and women
Anti-sperm antibodies
anti-sperm antibodies can be detected through:
semen, cervical mucosa or serum
detects the presence of IgG antibodies of anti-sperm antibodies
Mixed agglutination reaction test
more specific procedure that detects IgG, IgM, and IgA antibodies of anti-sperm antibodies
Immunobead test
(semen analysis) Microbial and Chemical Testing: indication of infection is detected through the presence of
> 1.0 million/mL (leukocytes)
Microbial testing for semen analysis involves routine aerobic and anaerobic cultures and tests that usually detects:
chlamydia tracomatis
mycoplasma hominis
and ureaplasma urealyticum
Reference value of chemical testing for semen analysis:
Neutral-a-glucosidase
> =20 uml/ejaculate
Reference value of chemical testing for semen analysis: Zinc
> = 2.4 umol/ejaculate
Reference value of chemical testing for semen analysis: Citric acid
> = 52 umol/ejaculate
Reference value of chemical testing for semen analysis:
Acid ejaculate
Glutamyl Transpeptidase
≥ 200 units/ ejaculate
Reference value of chemical testing for semen analysis:
Fructose
≥ 13 umol/ejaculate
Indication (↓ levels) of chemical testing for semen analysis:
Neutral-a-glucosidase
Glycerophosphocoline
L-carnitine
Disorder of epididymis
Indication (↓ levels) of chemical testing for semen analysis:
Zinc
Citric Acid
Acid phosphatase
Glutamyl transpeptidase
Lack of prostatic fluid
Indication (↓ levels) of chemical testing for semen analysis:
Fructose
lack of seminal fluid
Cutting of vas deferens
vasectomy
Post Vasectomy Semen Analysis detects:
presence or absence of spermatozoa
Sperm function test
Hamster Egg Penetration
Cervical Mucus Penetration
Hypo-Osmotic Swelling
In Vitro Acrosome Reaction
Sperm are incubated with species nonspecific hamster eggs and penetration is observed microscopically
Hamster egg penetration
Observation of sperm’s ability to penetrate partner’s midcycle cervical mucus
Cervical Mucus Penetration
Sperm exposed to low sodium concentrations are evaluated for membrane integrity and sperm viability
Hypo-osmotic Swelling
Hypoosmotic swelling indication of normal cell morphology
Swelling = normal
Evaluation of Acrosomoe to produce enzymes essential for ovum penetration
In Vitro Acrosome Reaction
