Hospital Corpsman 3 &
2: June 1989
Chapter 6: Clinical Laboratory
Naval Education and Training Command
Introduction
A basic knowledge of clinical laboratory procedures is
required of all hospital corpsmen, particularly those working at
small dispensaries and isolated duty stations without the
supervision of a medical officer. The patient's complaint may be
of little value by itself, but coupled with the findings of a few
easily completed laboratory studies, a diagnosis can usually be
surmised and treatment initiated.
Hospital corpsmen who can perform blood and urine tests
and interpret the results are better equipped to determine the
cause of illness or to request assistance, since they can give a
more complete clinical picture. Consequently, their patients can
get treated sooner.
In this chapter we will discuss blood collection, the
microscope, and step-by-step procedures for the complete blood
count and basic urinalysis.
Blood Collection
The two principal methods of obtaining blood samples are
finger puncture and venipuncture. Both methods have their
advantages and disadvantages, but for most clinical examinations,
blood is best obtained from a vein.
Finger Puncture
The finger puncture is used when a patient is burned
severely or is bandaged so that the veins are either covered or
inaccessible. It is also used when only a small amount of blood
is needed.
Equipment Required
Arrange your equipment in an orderly manner and
have it within easy reach. As with many other laboratory
procedures, wash your hands prior to the procedure.
Procedure
-
Using the middle or ring finger, massage or "milk"
the finger down towards the fingertip. Repeat this
"milking" five or six times.
-
Cleanse the fingertip with an alcohol pad or
Povidone-iodine solution and let dry. 3. Take a lancet
and make a quick deep stab on the side of the finger
(off-center). To obtain a large rounded drop, the
puncture should be across the striations of the fingertip
(fig. 6-1).
-
Wipe away the first drop of blood to avoid dilution
with tissue fluid. Avoid squeezing the fingertip to
accelerate bleeding as this tends to dilute the blood
with excess tissue fluid, but gentle pressure some
distance above the puncture site may be applied to obtain
a free flow of blood.
-
When the required blood has been obtained, apply a
pad of sterile gauze and instruct the patient to apply
pressure, then apply a bandage.
When dealing with infants and very small children,
the heel or great toe puncture is the best method to obtain
a blood specimen. It is performed in much the same
way.
Venipuncture (Vacutainer Method)
The collection of blood from a vein is called
venipuncture. For the convenience of technician and patient,
arm veins are best for obtaining a blood sample. If arm veins
cannot be used due to bandages, IV fluid therapy, thrombosed or
hardened veins, etc., consult your supervisor for instructions
on the use of hand or foot veins. DO NOT DRAW BLOOD FROM AN ARM
WITH IV FLUID RUNNING INTO IT. CHOOSE ANOTHER SITE. THE FLUID
ALTERS TEST RESULTS.
Equipment required
-
Sterile gauze pads (2 x 2)
-
70 percent isopropyl alcohol or Povidone-iodine
solution
-
Tourniquet
-
Vacutainer needles and holder
-
Vacutainer tubes appropriate for the test to be
performed
Position the patient so that the vein is easily
accessible and you are able to perform the venipuncture in a
comfortable position. Always have the patient either lying
in bed or sitting in a chair with the arm propped up. NEVER
PERFORM A VENIPUNCTURE WITH THE PATIENT STANDING UP, AND USE
CAUTION TO ENSURE THE PATIENT DOES NOT FALL FORWARD FROM HIS
OR HER SEAT.
Procedure
-
Wash hands.
-
Assemble equipment.
-
Explain the procedure to the patient.
-
Apply the tourniquet around the arm approximately 2
to 3 inches above the anticubital fossa with enough
tension so that the VEIN is compressed but not the
ARTERY. A sphymomanometer may be used instead of a
tourniquet if a patient is difficult to draw.
-
Position the patient's arm extended with little or no
flexion at the elbow.
-
Locate a prominent vein by palpation (feeling). If
the vein is difficult to find, it may be made more
prominent by massaging the arm with an upward motion to
force blood into the vein.
-
Cleanse the puncture site with a 70 percent alcohol
pad or Povidone-iodine solution and allow to dry.
CAUTION: After cleaning the puncture site, only
the sterile needle should be allowed to touch
it.
-
"Fix" or hold the vein taut. This is best
accomplished by placing the thumb under the puncture site
and exerting a slight downward pressure on the skin or
placing the thumb to the side of the site and pulling the
skin taut laterally. (See figure
6-2). 9. Using a smooth continuous motion, introduce
the needle into the side of the vein at about a 15 degree
angle with the skin (fig.
6-2). (Bevel of the needle should be up.)
-
Holding the vacutainer barrel with one hand, push the
tube into the holder with the other hand and watch for
the flow of blood into the tube until filling is
completed.
-
While holding the vacutainer with one hand, release
the tourniquet with the other.
-
Place a sterile gauze over the puncture site and
remove the needle with a quick, smooth motion.
-
Apply pressure to the puncture site and instruct the
patient to keep the arm in a straight position. Have the
patient hold pressure for at least 3 minutes.
-
Take this time to invert any tubes that need to have
anticoagulant mixed with the blood, then label the
specimens.
-
Reinspect the puncture site and apply a bandage.
The Microscope
Before any attempts are made to view blood smears,
urinary sediments, bacteria, parasites, etc., it is absolutely
essential that the beginner know the instrument with which he or
she will be spending considerable time-the microscope (fig.
6-3). The microscope is a precision instrument used repeatedly
in many areas of the medical laboratory to make visible those
objects that are too small to be seen by the unaided eye. This is
accomplished by means of a system of lenses of sufficient
magnification and resolving power (ability to show, separate, and
distinguish) so that small elements lying close together in a
specimen appear larger and distinctly separated. Most laboratories
are equipped with binocular (two-eye-piece) microscopes, but
monocular microscopes are also commonly used. The microscope most
often used in the laboratory is a compound microscope that
consists of the various pieces identified and discussed briefly
below:
Framework:
Base-structure on which the microscope rests.
Arm-structure that supports the magnification and
adjustment system; it is the handle by which the microscope is
carried.
Stage-platform on which a preparation is placed for
examination. In the center of the stage is the aperture or hole
to allow passage of light from the condenser. Mechanical
stage-means by which the preparation may be moved about on the
stage.
Illumination System:
Mirror-usually double, a flat surface on one side,
and a concave surface on the other side. The concave surface is
used in the absence of a condenser. Many microscopes have a
built-in light source instead of a lamp and mirror. Internal
light source-built into the base of the microscope, and
provides a more precise steady source of light into the
microscope.
Condenser-composed of a compact lens system located
between the mirror and stage. The condenser (usually an Abbe
condenser) concentrates (condenses) the light through the
aperture in the stage to the objective lens.
Iris diaphragm-controls the amount of light reaching
the condenser. The size of the iris diaphragm opening should
approximate that of the face of the objective lens. Thus, as a
general rule, the diaphragm is completely closed when
liquid.preparations are observed with the low-power objective,
and wide open when stained preparations are observed with the
oil-immersion lens using natural light.
Magnification System:
Revolving nosepiece-contains openings into which
objective lenses may be fitted and that may be revolved to
bring an objective into the desired position.
Objective lenses-usually a set of three consisting of
a low-power lens (approximate focus 16 mm, magnification 10X),
a highpower lens (approximate focus 4 mm, magnification 45X),
and an oil-immersion lens (approximate focus 1.8 mm,
magnification 100X). Numerical aperture (NA) refers to the
angle of the maximum cone of light that may enter the
objective. The greater the numerical aperture, the greater the
resolution, or ability of the microscope to separate small
details clearly.
The body tube-through which light passes from the
objective to the ocular lens. The ocular lenses
(eyepieces)-usually a 10X is provided: the number indicates the
magnification (in diameters) produces by the ocular of the
image formed by the objectives. Magnification is determined by
the ratio between the size of the virtual image and the real
size of the object. It is expressed in diameter multiples, for
example 100X. By multiplying the magnification engraved on the
objective by that engraved on the eyepiece, one can determine
the total magnification. The total magnification resulting from
the systems of lenses is determined by the combination of
objectives and oculars:
Objective lens
|
Color Code
|
l0X Ocular
|
Total Magnification
|
16 mm-l0X
|
green
|
l0x
|
lOOX
|
4 mm-45X
|
yellow
|
l0x
|
450X
|
1.8 mm-100X
|
red
|
l0X
|
1OOOX
|
Adjustment System
Composed of two parts, both of which
raise or lower the body tube together with the lens system.
Coarse adjustment-the larger and innermost knob; by
rotating the control knob, the image appears and is in
approximate focus.
Fine adjustment-the smaller and outermost knob; by
rotating this control knob, it renders the image clear and
well-defined.
Focusing the Microscope
The process of focusing consists of adjusting the
relations between the optical system of the microscope and the
object to be examined so that a clear image of the object is
obtained. The distance between the upper surface of a glass
slide on the microscope stage and the faces of the objective
lens varies according to which of the three objectives is in
focusing position. Thus, the intervening distance with the
low-power objective (l0X) is the greatest (16 mm), that for the
oil-immersion lens (l00X) is the smallest (1.8 mm), and that
for the high-power objective (45X) is intermediate (4 mm). As a
result, the focusing operation must be conducted with skill to
avoid damage to the objective lens, the specimen, or both. It
is good practice to obtain a focus with the low-power objective
first, then change to the higher objective required. Most
modern microscopes are equipped with parfocal objectives, which
means that if one objective is in focus, the others will be in
approximate focus when the nosepiece is revolved. With the
low-power objective in focusing position, observe the following
steps in focusing.
-
Seated behind the microscope, lower your head to one
side of the microscope until your eyes are approximately at
the level of the stage.
-
Using the coarse adjustment knob, lower the body tube
until the face of the objective is within 1/4 inch of the
object. Most microscopes are constructed in such a manner
that the low-power (green) objective cannot be lowered to
make contact with the object on the stage.
-
While looking through the ocular, use the coarse
adjustment knob to elevate the body tube until the image
becomes visible. Then use the fine adjustment knob to obtain
a clear and distinct image. Do not move the focusing knob
while changing lenses.
-
If the high-power objective (yellow) is to be used next,
bring it into position by revolving the nosepiece (a
distinct "click" indicates it is in proper alignment with
the body tube). Use the fine adjustment knob only to bring
the object into exact focus. Of course, light adjustment
must be made; open the iris diaphragm of the condenser to
accommodate more light.
-
The oil-immersion objective (red) is used for detailed
study of stained blood and bacterial smears. Remember that
the distance between objective lens and object is very
short, and great care must be employed. After focusing with
the highpower objective 'and scanning for well defined
cells, raise the objective, place a small drop of immersion
oil, free of bubbles, on the slide, centering the drop in
the circle of light coming through the condenser. Next,
revolve the nosepiece to bring the oil-immersion objective
into place, and by means of the coarse adjustment knob,
slowly lower the body tube until the lens just makes contact
with the drop of oil on the slide. The instant of contact is
indicated by a flash of light illuminating the oil. The
final step in focusing is done with the fine adjustment
knob. It is with this lens in particular that lighting is
important; the final focus, clear and well-defined, will be
obtained only when proper light adjustment is made.
Care of the Microscope
The microscope is an expensive and delicate
instrument that should be given proper care.
Moving or transporting the microscope should be done
by grasping the arm of the scope in one hand and supporting the
weight of the scope with the other hand. Avoid sudden jolts and
jars.
Make sure the microscope is kept clean at all times;
when not in use, enclose it in a dustproof cover or store in
its case. Remove dust with a camel hair brush. Lenses may be
wiped carefully with lens tissue.
When the oil immersion lens is not being used, remove
the oil with lens tissue. Use oil solvents, such as xylol, on
lenses only when required to remove dried oil and only in the
minimal amount necessary. Never use alcohol or similar solvent
to clean lenses.
Complete Blood Count
The complete blood count consists of:
-
Total red blood cell count (RBC)
-
Hemoglobin determination
-
Hematocrit reading
-
Total white blood cell count (WBC)
-
Differential white blood cell count
Red Blood Cell (Erythrocyte) Count
The red blood cell count is made to determine the
number of red cells in one cubic millimeter (mm3) of blood. The
normal red blood cell count is as follows:
-
adult male 5.4 ± .8 million/mm3
-
adult female 4.8 ± .6 million/mm3
-
newborn 5.1 ± .9 million/mm3
A lower count is usually a sign of
anemia.
A lower count is usually a sign of
anemia.
Manual Sahli Pipette Method
The materials listed below are required to perform
a red blood cell count using the manual Sahli pipette
method:
Procedure
-
Using well mixed anticoagulated blood or blood
directly from a fingerstick, fill the clean, dry, red
cell pipette (fig. 6-4)
exactly to the 0.5 mark with the aid of the suction tube.
Hold the pipette in a nearly horizontal position so that
the exact level of the blood can be seen. The curve of
the tip may rest on the skin, but the orifice must be
free to immerse in the drop of blood to avoid air
bubbles. If the blood level exceeds the 0.5 mark,
withdraw excess blood by touching the tip to the skin
surface. Do not touch it to gauze or cotton since these
material absorb the fluid portion of the blood, leaving
behind a much higher concentration of cells.
-
Wipe the blood from the outside of the pipette,
taking care not to touch the very tip. Immerse the tip in
the RBC diluting fluid and aspirate fluid exactly to the
101 mark, slightly rotate the pipette while doing so. It
is best to hold the pipette in an almost vertical
position to avoid formation of air bubbles in the bulb.
DO NOT DELAY BETWEEN STEPS 1 AND 2. IF THE BLOOD IS NOT
DILUTED PROMPTLY, IT WILL DRY IN THE PIPETTE. Start to
draw diluting fluid into the pipette as soon an the tip
of the pipette is immersed in fluid to avoid loss of
blood cells. Wipe the excess diluting fluid from the
pipette, taking care not to touch the very tip. Filter
the diluting fluid regularly to remove accidentally
introduced blood cells.
-
Remove the suction tube and shake the pipette vigorously for 3
minutes. DO NOT SHAKE IN THE DIRECTION OF THE LONG AXIS. (See figure
6-5.)
-
Discard the clear fluid (about three drops) from the
stem of the pipette. The counting chamber (figure
6-6) must be loaded with fluid from the pipette's
bulb.
-
Place the coverglass on the counting chamber, making
sure both are clean and grease-free. (Fingerprints must
be completely removed.) Load the counting chamber by
touching the tip of the pipette against the edge of the
coverglass and the surface of the counting chamber
(figure 6-7). A properly
loaded counting chamber should have a thin, even film of
fluid under the coverglass. Allow 3 minutes for cells to
settle. If fluid flows into the grooves (moats) at the
edges of the chamber or if air bubbles are seen in the
field, the chamber is flooded and must be cleaned with
distilled water, dried with lens tissue, and reloaded. If
the chamber is underloaded, carefully add additional
fluid until properly loaded.
-
Place the hemacytometer (figure
6-8) on the microscope. Use the low-power lens to
locate the five small fields (1, 2, 3, 4, and 5) in the
large center square bounded by the double or triple
lines. Each field measures 1/25 mm2, 1/10 mm in depth,
and is divided into 16 smaller squares. These smaller
squares form a grid that makes accurate counting
possible.
-
Switch to the high-power lens and count the number of
cells in field 1. Move the hemacytometer until field 2 is
in focus and repeat the counting procedure. Continue
until the cells in all five fields have been counted.
Note that the fields are numbered clockwise around the
chamber, with field 5 being in the center. Count the
fields in this order. To count the cells in each field,
start in the upper left small square and follow the
pattern indicated by the arrow in figure
6-8. Count all of the cells within each square,
including cells touching the lines at the top and on the
left. DO NOT COUNT ANY OF THE CELLS TOUCHING THE LINES ON
THE RIGHT AND AT THE BOTTOM.
-
Total the number of cells counted in all five fields
and multiply by 10,000 to arrive at the number of red
cells per cubic millimeter of blood. The number of cells
counted in each field should not vary by more than 20. A
greater variation may indicate poor distribution of the
cells in the fluid, resulting in an inaccurate count.
-
Immediately after completing the count, clean the
counting chambers with distilled water and dry it with
lens tissue. Rinse pipettes first with cold water, then
with acetone. Draw air through the pipette until it is
dry. The pellet should move freely in the bulb if the
pipette is dry.
Some common sources of error are:
-
Improper dilution-not drawing blood exactly to the
0.5 mark or using too much diluting fluid
-
Dirty equipment-diluting fluid unfiltered; greasy
glassware; dirty microscope; wet pipettes
-
Poor mixing or not discarding first few drops of
fluid
-
Poorly loaded counting chamber
-
Chipped pipettes. Discard pipettes with chipped or
broken tips.
-
Use of gauze, cotton, or filter paper to remove
excess blood from the pipette.
Uniopette Method
The Uniopette disposable diluting pipette system
for the red blood cell count provides a convenient, precise,
and accurate method for obtaining a red blood cell count.
The disposable kit consists of a shielded capillary pipette
(10 microliter (ul) capacity) and a plastic reservoir
containing a premeasured volume of diluent (1:200
dilution).
Procedure
-
Using the shield on the capillary pipette, puncture
the diaphragm in the neck of the reservoir with the tip
of the capillary shield.
-
After obtaining free-flowing blood from a lancet
puncture of the finger, remove the protective plastic
shield from the capillary. Holding the capillary slightly
above the horizontal, touch the tip to the blood source.
Capillary action will fill the tube until blood
collection stops automatically, e.g., when the proper
amount (10 ul) has been obtained. Wipe blood off the
outside of the capillary tube, making sure none is
removed from inside the capillary tube. An alternative
source of blood is a thoroughly mixed fresh venous blood
sample obtained by venipuncture.
-
Squeezing the reservoir slightly, cover the upper
opening of the capillary overflow chamber with the index
finger and seat the capillary tube holder in the
reservoir neck. Release pressure on the reservoir and
remove the finger from the overflow chamber opening.
Suction will draw the blood into the diluent in the
reservoir.
-
Squeeze the reservoir gently two or three times to
rinse the capillary tube, forcing diluent into-but not
out of-the overflow chamber, releasing pressure each time
to return diluent to the reservoir. Close the upper
opening with your index finger and invert the unit
several times to mix the blood sample and the
diluent.
-
For specimen storage, cover the overflow chamber of
the capillary tube with the capillary shield.
-
Immediately prior to cell counting, mix adequately
again by gentle inversion, taking care to cover the hole
with your index finger.
-
Remove the pipette from the reservoir. Squeeze the
reservoir and reseat the pipette in the reverse position,
releasing pressure to draw any fluid in the capillary
tube into the reservoir. Invert and fill the capillary
tube by gentle pressure on the reservoir. After
discarding the first 3 drops, charge the counting chamber
of the hemacytometer by gently squeezing the
reservoir.
-
Using the high power objective, count the red blood
cells in the red blood cell counting areas.
-
Calculations: Multiply the number of cells counted by
10,000 to obtain the red cell count.
Example:
|
The number of cell in the 5 counting squares
was 423.
|
|
|
|
|
|
The cell count
|
=
|
423 x 10,000
|
|
|
=
|
4,230,000
|
Hemoglobin Determination
Of the many methods of hemoglobin estimation, the
most accurate is reading of hemoglobin as oxyhemoglobin in the
photometer, after dilution of the blood with a weak alkali. The
Haden-Hausse, Sahli-Hellige, and Newcomer tests, based on acid
hematin formed by the action of hydrochloric acid on
hemoglobin, are sufficiently accurate for routine examination,
provided they are properly done. Since relatively few ships and
stations are equipped with a photometer, we will discuss the
Sahli-Hellige method.
Materials Required for Sahli-Hellige Test
Procedure
-
With a medicine dropper, place 5 drops of the 0.1N
HCl in the bottom of the graduated tube. Place the tube
in the color comparator.
-
Using well-mixed venous blood or fingertip blood,
fill the pipette to the 20 mm3 mark.
-
Wipe blood from the outside of the pipette. Transfer
blood to the Sahli tube. Note the time.
-
Aspirate distilled water into the pipette two or
three times and transfer these washings to the tube.
-
Shake until the blood is well mixed and the tube is a
uniform color.
-
Add distilled water, drop by drop, each time mixing
the solution with the stirring rod. Keep adding water and
mixing until the color of the solution matches the
standards on either side. Remove the stirring rod from
the tube each time before comparing. Natural light makes
more accurate readings possible.
-
Five minutes after time noted, read the result from
the scale on the tube by noting the graduation mark at
the lower edge of the meniscus. Read and report both scales.
Reporting. Findings are reported both in grams per
100 ml of whole blood and in percentages of normal values.
There are a number of modifications of the Sahli-Hellige
method, and 100 percent may be equal to from 13.8 g to 17.3
g. In the sets usually used in the Navy, however, 100
percent is equal to 14.5 g of hemoglobin per 100 ml of whole
blood. After reading the percentage on the scale, turn the
tube and read from the other side to get the equivalent
reading in grams.
If either scale is hard to read, remember that
100% / 14.5 g = 6.9, so one gram of hemoglobin is equal to
6.9 percent. If only one scale can be read, the other
reading can be computed.
Caution: Equipment must be clean and dry before
determination is started. Wipe all blood from the outside of
the pipette before you insert it into the tube. Twenty cubic
millimeters is a small volume, and a few blood cells
clinging to the outside of the pipette can cause a
significant error in findings.
Hematocrit (Packed Cell Volume)
Determination
Hematocrit is the volume of erythrocytes expressed as
a percentage of the volume of whole blood in a sample. The
venous hematocrit agrees closely with the hematocrit obtained
from a skin puncture; both are greater than the total body
hematocrit. Dried heparin, balanced oxalate, or EDTA is
satisfactory as an anticoagulant.
Although the microhematocrit method is not available
at all duty stations, it is the most accurate means of
determining blood volume and should be used whenever feasible.
This test is rapidly replacing the red cell count for general
purposes since it is easier, quicker, and more accurate. The
method described here is the microhematocrit method.
Normal Values. The normal hematocrit for males is 42
to 50 percent, for females, 40 to 48 percent. A value below an
individual's normal range for sex and age indicates anemia.
Materials Required
-
Capillary tubes, plain or heparinized
-
Modeling clay sealant or microburner
-
Microhematocrit centrifuge
-
Microhematocrit reader
Procedures
-
Fill the capillary tube two-thirds to three- quarters
full with well-mixed, oxalated venous blood or fingertip
blood (for fingertip blood use heparinized tubes, and
invert several times to mix).
-
Seal one end of the tube with clay or seal the empty
end of the tube in a small flame of a microburner.
-
Place the filled tube in the microhematocrit
centrifuge, with the plugged end away from the center of
the centrifuge.
-
Centrifuge at a preset speed of 10,000 to 12,000 rpm
for 5 minutes. If the hematocrit exceeds 50 percent,
centrifuge for an additional 3 minutes.
-
Place the tube in the microhematocrit reader. Read
the hematocrit by following the manufacturer's
instruction on the microhematocrit reading device.
White Blood Cell (Leukocyte) Count
The total white cell count
determines the number of
white cells per cubic millimeter of blood. A great deal of
information can be derived from white cell studies. The white
cell count and the differential count are common laboratory
tests and almost a necessity in determining the nature and
severity of systemic infections.
Normal Values. The normal range is 4,000 to 11,000
cells per cubic millimeter of whole blood.
Abnormal White Blood Cell Count
-
Leukocytosis (abnormally high count). This may be
caused by:
-
Systemic or local infections (usually due
to bacteria). These counts are highly variable and not
diagnostic. Some infections and representative white
cell counts are:
-
pneumonia-20,000 to 30,000/mm3
-
meningitis-20,000 to 30,000/mm3
-
appendicitis-10,000 to 30,000/ mm3
-
Dyscrasia of blood-forming tissues. This is not
caused by any known bacteria, but is due to a
malfunctioning of the marrow and lymph tissues,
resulting in extremely high white cell counts, which
sometimes exceed 1,000,000/mm3. This is commonly known
as leukemia, or blood cancer.
-
Physiologic conditions, with counts as high as
15,000/mm3. Some of these may occur as follows:
-
in the newborn
-
during late pregnancy
-
during labor
-
accompanying severe pain
-
after exercise or meals
-
after cold baths
-
during severe emotional upset
-
Leukopenia (abnormally low count). This may be caused
by:
-
Severe or advanced bacterial infections,
such as typhoid, paratyphoid, and sometimes tularemia,
or when the bacterial infection has been undetected
for a period of time as with chronic beta
streptococcal infections of the throat.
-
Infections caused by viruses and rickettsiae, such
as measles, rubella, smallpox (until the 4th day),
infectious hepatitis, psittacosis, dengue,
tsusugamushi fever, and influenza (when it may fall to
1,5000/mm3, or shift to leukocytosis if complications
develop).
-
Protozoal infections (such as malaria) and
helminthic infections (such as Trichinosis). In
malaria, slight leukocytosis may develop for a short
time during paroxysm, but shortly after its onset
leukopenia ensues. With trichinosis there may be
leukocytosis with an increase in eosinophils (as high
as 50 to 70 percent).
-
Overwhelming infections when the body's defense
mechanism breaks down
-
Anaphylactic shock
-
Radiation
Manual Sahli Pipette Method
The materials listed below are required to perform
a white blood cell count using the manual Sahli pipette
method:
The gentian violet is not necessary, but by
staining the nucleus, it makes the cells more refractile
and helps to make an accurate count.
Procedure
-
Draw well-mixed anticoagulated venous blood or
fingertip blood to the 0.5 mark on the white cell
pipette.
-
Observing the same precautions as for red cell count,
draw diluting fluid to the 11.0 mark.
-
Shake the pipette for 3 minutes. Do not shake along
the long axis. (See figure
6-5.)
-
Load the counting chamber, using the same technique
as for the red cell count.
-
Count the white cells with the low-power lens in each
of the four large corner fields (A, B, C, and D, in figure 6-8). Use subdued
lighting. Go clockwise around the counting chamber; that
is, from field A to field B to field C to field D. For
convenience each field is divided into 16 smaller
squares. Starting with the small square in the upper left
corner of the field, count the cells in each square in
the top row, moving across the field to the right, then
drop down one row of squares and work back to the left,
as indicated by the arrow in figure 6-8. Remember the
rule for counting cells: COUNT THE CELLS TOUCHING THE
BORDER LINES AT THE TOP AND ON THE LEFT. DO NOT COUNT THE
CELLS TOUCHING THE LINES ON THE RIGHT AND AT THE
BOTTOM.
-
When all the cells in the four fields have been
counted, multiply the count by 50 for the total white
cell count.
-
Immediately after completing the count, clean the
counting chamber with distilled water and dry it with
lens tissue. Rinse pipettes first with cold water, then
with acetone. Draw air through the pipette until it is
dry. The pellet should move freely in the bulb if the
pipette is dry. Sources of error. The errors are
generally caused by the same mistakes as described for
red cell counts.
Uniopette Method
The Uniopette disposable kit for doing a white
blood cell count consists of a shielded capillary pipette
(20 ul capacity) and a plastic reservoir containing a
premeasured volume of diluent (1:100 dilution).
Procedure
-
Using the shield on the capillary pipette, puncture
the diaphragm in the neck of the reservoir with the tip
of the capillary shield.
-
After obtaining free-flowing blood from a lancet
puncture of the finger, remove the protective plastic
shield from the capillary. Holding the capillary slightly
above the horizontal, touch the tip to the blood source.
Capillary action will fill the tube until the blood
collection stops automatically, e.g., when the proper
amount (20 ul) has been obtained. Wipe blood off the
outside of the capillary tube, making sure none is
removed from inside the capillary tube. An alternative
source of blood is a thoroughly mixed fresh venous blood
sample obtained by venipuncture.
-
Squeezing the reservoir slightly, cover the upper
opening of the capillary overflow chamber with your index
finger and seat the capillary tube holder in the
reservoir neck. Release pressure on the reservoir and
remove your finger from the overflow chamber opening.
Suction will draw blood into the diluent in the
reservoir.
-
Squeeze the reservoir gently two or three times to
rinse the capillary tube, forcing diluent into-but not
out of-the overflow chamber, releasing pressure each time
to return diluent to the reservoir. Close the upper
opening with your index finger and invert the unit
several times to mix the blood sample and diluent.
-
For specimen storage, cover the overflow chamber of
the capillary tube with the capillary shield.
-
Immediately prior to cell counting, mix adequately
again by gentle inversion, taking care to cover the hole
with your index finger.
-
Remove the pipette from the reservoir. Squeeze the
reservoir and reseat the pipette in the reverse position,
releasing pressure to draw any fluid in the capillary
tube into the reservoir. Invert and fill the capillary
tube by gentle pressure on the reservoir. After
discarding the first 3 drops, charge the counting chamber
of the hemacytometer by gently squeezing the
reservoir.
-
Using the high power objective, count the white blood
cells in the nine large squares
-
Calculation: Add 10 percent to the number of cells
counted in the nine large squares and multiply by 100 to
obtain the white cell count.
Example: The number of cells in 9 large
squares was 90.
|
|
|
|
|
|
The cell count
|
=
|
[90 + (0.1 x 90)] x 100
|
|
|
|
|
|
|
=
|
[90 + 9] x 100
|
|
|
=
|
99 X 100 = 9900
|
Differential White Blood Cell
Count
The total white cell count is not necessarily
indicative of the severity of a disease, since some serious
ailments may show a low white cell count. However, the
percentage distribution of the different types of leukocytes in
the blood often provides more helpful information in
determining the severity and extent of the infection than any
other single procedure used in the examination of the blood.
The differential count gives these percentages.
Normal Values. The normal percentages of the
different leukocytes are:
Eosinophils
|
(Eos)
|
2-4 percent
|
Basophils
|
(Basos)
|
0-2 percent
|
Lymphocytes
|
(Lymphs)
|
21-35 percent
|
Monocytes
|
(Monos)
|
4-8 percent
|
Neutrophils
|
(Neuts)
|
|
Metamyelocytes
|
(Metas)
|
0 percent
|
Bands or Stab forms
|
(Bands)
|
0-10 percent
|
Segmented
|
(Segs or Polys)
|
51-67 percent
|
Most hospital corpsmen have heard the expression
"shift to the left" and "shift to the right." These terms are
often loosely used in refer best be explained by the following
diagram:
Percentage Distribution of the Different
Leukocytes
|
|
EOS
|
BASOS
|
METAS
|
BANDS
|
SEGS
|
LYMPHS
|
MONOS
|
|
|
|
|
|
|
|
|
Normal Percent
|
2-4
|
0-2
|
0
|
0-10
|
51-67
|
21-35
|
4-8
|
The metamyelocytes, bands, and segmented neutrophils
constitute the neutrophilic cells. When the cells to the left
of the segs are increases, it is a "shift to the left." If the
segmented neutrophils increase, it is a "shift to the right."
The true "right shift" will show numerous hypersegmented
(having six or more lobes) neutrophils.
General Interpretations of Leukocytic Changes
The severity of an infection may be determined by
the total white cell count and the differential count.
-
Leukocytosis (abnormally high white cell count) with
an increase in the percentage of neutrophil indicates a
severe infection with a good response of the bone marrow.
The primary phagocytes (bacteria-destroying cells) are
the neutrophils, and the bone marrow should supply large
numbers of these to combat the infection. The greater the
"shift to the left" (increase in immature neutrophils),
the more severe the infection. The appearance of numerous
juvenile cells (metamyelocytes) indicates irritation of
the bone marrow with regeneration. If the infection
continues and the patient's resistance declines, the
shift advances further to the left. If improvement
ensues, the shift declines and recedes to normal.
-
A falling white cell count with the number and
maturity of neutrophils progressing toward normal
indicates recovery.
-
A continued "shift to the left" with a falling total
white cell count indicates a breakdown of the body's
defense mechanism and is a poor prognosis.
-
The percentage of eosinophils, lymphocytes, and
monocytes generally decreases in acute infections.
-
In tuberculosis an increase in monocytes
(monocytosis) indicates activity in the infected area. An
increase in lymphocytes (lymphocytosis) indicates
healing.
-
Eosinophils increase in parasitic infections and
allergic conditions.
Materials Required for Differential Count
-
Four plain glass microscope slides, clean and
dry.
-
Wright's stain, powder or tablet form. Prepare this
stain as follows.
-
Tablets: Use one tablet for each 10 ml of reagent
grade methyl alcohol. Dissolve tablets after crushing
them with the aid of a clean, dry mortar and pestle.
Pour the solution in a stopper bottle and store for 30
days in a dark place, shaking it periodically during
this period. Filter before use. Fill a dropper bottle
for use at the lab bench.
-
Powder (preferred): Use 0.1 g of powder for each
60 ml of reagent grade methyl alcohol. Prepare in the
same manner as the tablets. Fill a dropper
bottle.
-
Buffering solution. This may be either:
-
Distilled water with a pH of 7. Use from a
dropper bottle.
-
A prepared buffer solution with a pH of 6.4.
Prepare this by dissolving 6.63 g monobasic
potassium phosphate and 3.20 g of dibasic sodium
phosphate in 1,000 ml of distilled water.
-
Staining rack
-
Mouthpiece and tube as used in blood counting
pipettes
-
Microscope and lamp
-
Immersion oil
-
Blood cell counter
Technique for Making Smears
-
Cleanse finger with 70 percent isopropyl alcohol and
puncture it as described previously.
-
Wipe off the first drop of blood with dry, sterile
gauze.
-
When a second drop forms, touch it lightly with a
clean, grease-free slide. Place the slide on a flat
surface with the drop of blood up. A smear can also be
made with a drop of blood from a needle or collecting
tube.
-
Hold a second slide between thumb and forefinger and
place the edge at a 45 degree angle against the top of
the slide, holding the drop of blood. Back the second
slide down until it touches the drop of blood. The blood
will distribute itself along the edge of the slide in a
formed angle.
-
Push the second slide along the surface of the other
slide, drawing the blood across the surface in a thin,
even smear. If this is done with uniform rapidity and
without wobbling the slide, a good smear will result. Try
to keep the blood from reaching the extreme edges of the
slides. Large cells have a tendency to stack up on the
perimeter of the smear, and letting the smear reach the
edges of the slide will aggravate this tendency. The
smear should show no wavy lines or blank spots (see figure 6-9).
-
Let the smear air-dry.
Technique for Staining Smears
-
Place the smear on a staining rack. Flood it with
about 1 ml of Wright's stain and allow it to stand for 2
minutes.
-
Add an equal quantity of buffer. There should be no
run-off of fluid from the slide. A little experimentation
will show just how much stain and buffer the slide will
hold. Mix the buffer and stain by blowing air through the
rubber pipette tube and directing the current of air
about the surface of the slide. Mix until a metallic
(copper looking) film appears. Let stand for 3
minutes.
-
Wash with tap water, provided pH testing has shown
the water to be neutral. If the tap water is not neutral,
wash the slide with distilled water. The stain and the
metallic film must be floated off to prevent streaking,
so keep the slide flat and horizontal in the stream of
water. If the slide is tilted, the metallic film will
settle to the surface of the smear and remain there.
-
Wipe the stain from the bottom of the slide. Let it
air-dry. A good smear should be thin and evenly
distributed, and it must be dry before staining. Always
make at least two smears for each patient, as the
additional smear should be examined to verify any
abnormal findings.
Staining times will vary with different batches of
Wright's stain. One batch may give best results with 3
minutes of staining and 2 minutes of buffering, another may
give better results with 2 minutes of staining and 3 minutes
of buffering. The only way to determine the best time
interval for a particular stain is by experimentation.
Next to incorrect time intervals, the most common
cause of poor results with Wright's stain is incorrect pH of
the staining fluid. If the stain is too acid, the red cells
in the smear will stain a bright pink or may even be
colorless, while an alkaline stain will cause the red cells
to appear blue-gray, with poor color definition. In either
case, the pH of the buffering solution should be
checked.
Technique for Differential Count
-
Place the stained slide on the microscope with a drop
of immersion oil and adjust the ocular lenses.
-
Using the oil immersion lens (red) (highest power),
scan the fields for areas where the red cells just touch.
In this area the blood smear is thinner, and
consequently, the white cells are easier to
identify.
-
Count 100 consecutive white cells (see figure
6-10 for identification), pressing the correct key on
the cell counter for each type of white cell
identified.
-
If you count 20 lymphocytes among the 100 cells, then
the patient has 20 percent lymphocytes. If an absolute
count has been requested, the total leukocyte count is
multiplied by individual percentages.
-
Example:
Patient has a white count of 8,000.
Differential count shows 20 percent
lymphocytes.
20 percent x 8,000 = 1,600
The patient has 1,600 lymphocytes per cubic
millimeter of blood.
Cell Identification
The ability to identify the different types of
white cells is not difficult to develop, but it does require
a thorough knowledge of staining characteristics and
morphology that can be gained only by extensive, supervised
practice. The beginner is likely to encounter some
difficulty in learning to differentiate between monocytes
and large lymphocytes, or between monocytes and atypical
(not typical) lymphocytes. It is possible also to confuse
eosinophils with basophils, since an alkaline stain may
cause the orange granules in an eosinophil to appear deep
blue or purple. If this happens, the reddish cast in the
granules can often be detected by judicious use of the minor
focusing knob. Also, the granulations of an eosinophil are
generally much finer than those of a basophil, and they tend
to concentrate in the cytoplasm.
The neutrophils are subclassified according to
age, and the age is indicated by the nucleus. If there is
any doubt as to the identity of the neutrophil, always
classify it with the older stage. For instance, if a single
irregularity in a horseshoe-shaped nucleus appears as a
break or a concentration of color, classify it as a
segmented neutrophil, not as a neutrophilic band. Practice
is the key. And remember-even experienced technicians often
disagree as to the identity of a particular cell.
If it is desirable to save a smear for
reexamination, remove the immersion oil by placing a piece
of lens tissue over the slide and moistening the tissue with
xylol. Draw the damp tissue across the slide and dry the
smear with another piece of lens paper.
A good smear (thin and evenly distributed) is
essential for accurate identification and counting.
The cytoplasm of an eosinophil contains numerous
coarse, reddish-orange granules, which are lighter colored
than the nucleus.
Scattered large, dark-blue granules, which are
darker than the nucleus, characterize the cell as a
basophil. Granules may overlay the nucleus as well as the
cytoplasm.
The cytoplasm of a lymphocyte is clear sky blue,
scanty, with few unevenly distributed, azurophilic granules
with a halo around them. The nucleus is generally round or
oval or slightly indented, and the chromatin is lumpy and
condensed at the periphery.
The largest of the normal white blood cells is the
monocyte. Its color resembles that of a lymphocyte, but its
cytoplasm is a muddy gray-blue. The nucleus is lobulated,
deeply indented or horseshoe-shaped, and has a relatively
fine chromatin structure. Occasionally the cytoplasm is more
abundant than in the lymphocyte.
The cytoplasm of a neutrophil has numerous fine
lilac-colored granules, which sometimes are hardly visible.
The nucleus is dark purple or reddish purple, and it may be
oval, horseshoe- or S- shaped, or segmented (lobulated). The
neutrophil is further subclassified according to age as:
-
Metamyelocyte (also called "juvenile"). This
is the youngest neutrophil generally reported. The
nucleus is fat, indented, and is usually "bean" shaped or
"cashew nut" shaped.
-
Band (sometimes called stab). This is the older or
intermediate neutrophil. The nucleus has started to
elongate and has curved itself into a horseshoe- or
S-shape. As the band ages, it progresses to:
-
Segmented. The nucleus is separated into two, three,
four, or five segments or lobes.
-
Hypersegmented. The nucleus is divided into six or
more segments or lobes.
Urinalysis
The physical and chemical properties of normal urine are
markedly constant; any abnormalities are easily detected. The use
of simple tests provides the physician with helpful information
concerning the diagnosis and management of many diseases.
This section will deal with the routine and microscopic
examination of urine specimens, some of the principles involved,
and some of the simpler interpretations of the findings.
Urine Specimens
Urine specimens for routine examinations must be
collected in aseptically clean containers. Unless circumstances
warrant, catheterization should be avoided because it may cause
urinary tract infections. Specimens of female patients are
likely to be contaminated with albumin and blood from menstrual
discharge, or with albumin and pus from vaginal discharge. For
bacteriologic studies, care must be taken to ensure that the
external genitalia have been thoroughly cleansed with soap and
water. The patient must then void the initial stream of urine
into the toilet or a suitable container and the remainder
directly into a sterile container. All urine specimens should
be examined when freshly voided or should be refrigerated to
prevent decomposition of urinary constituents and to limit
bacterial growth.
Random Specimen
This is a sample of urine voided without regard to
the time of day or fasting state. This sample is
satisfactory for most routine urinalyses. It is the least
valid specimen, since tests results may reflect a particular
meal or fluid intake.
First Morning Specimen
This is the first specimen of urine voided upon
rising. It is the best sample for routine urinalysis because
it is usually concentrated and more likely to reveal
abnormalities. If positive results are obtained from the
first morning specimen, the physician may order a 24-hour
specimen for quantitative studies.
Twenty-four Hour Specimen
This specimen measures the exact output of a
specific substance over a 24-hour period. To collect this
specimen:
-
Have the patient empty his or her bladder at 0800.
Discard this urine.
-
Collect all urine voided during the next 24
hours.
-
At 0800 the following day (end of 24-hour period),
instruct the patient to empty his or her bladder. Add
this urine to the pooled specimen.
-
Specimen must be refrigerated during collection.
-
Preservatives will be added to the first specimen
voided, according to the types of tests being
ordered.
Preservation of Specimens
To delay decomposition, use
-
Refrigeration. All specimens not being examined
immediately should be refrigerated.
-
Toluene. Simply add enough toluene to form a thin film
on the surface of the specimen. This film will prevent air
from reaching the urine. False positives are seldom
encountered with this preservative. Remember that the
toluene is on the surface, and all test samples must be
pipetted from BENEATH the surface.
-
Thymol. A small lump, floating on the surface, will
preserve a urine specimen for several days. Enough thymol
may dissolve to produce false positives for albumin. Do not
use more that 0.1g of thymol per 100 ml of urine.
Other common preservatives are formaldehyde, boric
acid, hydrochloric acid, and chloroform. The preservative used
must be identified on the label of the container. If no
preservative is used, it should be so stated.
Routine Examination
Volume (For 24-Hour Specimen or When Requested)
The normal daily urine volume for adults ranges
from 800 to 2000 ml, averaging about 1,500 ml. The amount of
urine excreted in 24 hours varies with fluid intake and the
amount of water lost through perspiration, respiration, and
bowel activity. Diarrhea or profuse sweating will reduce
urinary output; a high protein diet tends to increase it.
Daytime urine output is normally two to four times greater
than nighttime output.
Color
The normal color of urine varies from straw to
light amber. Diluted urines are generally pale; concentrated
urines tend to be darker. The terms used to describe the
color of urine are:
-
Colorless
-
Light straw
-
Straw
-
Dark straw
-
Light amber
-
Amber
-
Dark amber
-
Red
The color of urine may be changed by the presence
of blood, drugs, or diagnostic dyes. Examples are:
-
Red or red-brown (smokey appearance), due to the
presence of blood.
-
Yellow or brown (turning greenish with yellow foam
when shaken), due to the presence of bile.
-
Olive green to brown-black, caused by phenols.
-
Milky appearance, caused by chyle.
-
Dark orange, due to treatment with Pyridium.
-
Blue-green, due to methylene blue.
Transparency
Urine may be reported as clear, hazy, slightly
cloudy, cloudy, or very cloudy. Some physicians prefer the
term "turbidity" to "transparency," but both terms are
acceptable.
Freshly passed urine is usually clear or
transparent. In certain conditions it may be cloudy due to
the presence of blood, phosphates, crystals, pus, bacteria,
etc. A report of transparency is of value only if the
specimen is fresh. After standing, all urine becomes cloudy
due to decomposition, salts, and the action of bacteria.
Upon standing and cooling, all urine specimens will develop
a faint cloud composed of mucus, leukocytes, and epithelial
cells. This cloud settles to the bottom and is of no
significance.
Reaction
Normal urine is slightly acid but will become more
alkaline upon standing. The pH ranges from 4.6 to 8.0. The
acidity of urine is influenced by many factors, such as a
diet high in protein or fat, fasting and starvation, and
acid therapy. Alkaline urine may be produced by cystitis,
pyelonephritis, and sulfonamide therapy.
It is essential that an alkaline urine be
maintained during treatment with sulfonamides, since these
compounds are precipitated as crystals in acid solution. The
crystals will cause damage to the uriniferous tubules.
Sodium bicarbonate is generally used as an alkalizer.
Reaction to pH, protein, glucose, ketones,
bilirubin, blood, nitrite, and urobilinogen in urine may be
determined by the use of the Multistix and Color Chart. This
is a specially prepared multitest strip that is simply
dipped into the urine specimen and then compared with the
color values for the various tests on the accompanying
chart. The color chart indicates pH values, and the
numerical value should be reported.
Specific Gravity
The specific gravity of the specimen is the weight
of the specimen as compared to an equal volume of distilled
water. The specific gravity varies directly with the amount
of solids dissolved in the urine and normally is from 1.015
to 1.030 during a 24-hour period.
The first morning specimen of urine is more
concentrated and will have a higher specific gravity than a
specimen passed during the day. A high fluid intake may
reduce the specific gravity to below 1.010. In disease the
specific gravity of a 24-hour specimen may vary from as low
as 1.001 to as high as 1.060.
The specific gravity may be measured with the
urinometer or the index refractometer, available as standard
equipment at most duty stations. The refractometer may be
held manually (fig. 6-11) or
mounted on a stand like a microscope. The specific gravity
is determined by the index of light refraction through solid
material.
Measurement with Urinometer
-
Pour urine into the cylinder until it reaches a point
approximately 1 1/2 inches below the top of the cylinder.
Insert the urinometer, making sure that it is floating
freely. The cylinder should not overflow when the
urinometer is immersed. Read the bottom of the
meniscus.
-
If the specimen is cloudy, the urine should be
centrifuged before specific gravity readings are taken.
Cloudy urine tends to show low (and invalid) specific
gravity.
Measurement with Index Refractometer
-
Hold the index refractometer in one hand, and with
the other hand and two applicator sticks, place a drop of
urine on the glass section beneath the coverglass.
-
Hold the refractometer so that the light reflects on
the glass section and look into the ocular end. Read the
number that appears where the light and dark lines meet.
This is the specific gravity.
Glucosuria
Glucosuria is the presence of an abnormal amount of
glucose in the urine. Traces of sugar are often found in normal
urine, but are not enough to react to any of the routine tests
for glucose. However, different sugars in the urine are of
various clinical significance. When performing routine
urinalysis, we can test for glucose, and any trace of glucose
in the test suggests that something is wrong with the patient's
carbohydrate metabolism.
Methods for Measuring Glucose
-
Test Strip, Glucose and Blood. This test is specific
for glucose. Other sugars do not interfere.
-
Remove strip from container.
-
Dip into specimen. Remove and wait 30
seconds.
-
Immediately compare the test strip against the
color chart.
-
Record results. Normal urine is negative for
glucose.
-
If there is a positive reaction for glucose, a
Clinitest is done for both confirmation and
quantitation.
-
Urine-Sugar Test Tablets (Clinitest Tablets)
-
Place 5 drops of urine in a large test
tube.
-
Add 10 drops of water and mix well.
-
Add one Clinitest tablet.
-
Put the tube in a rack. Let stand until the
reaction is completed (foaming action has
stopped).
-
Compare the color with the color chart and record
the results.
-
If an orange color appears and then disappears,
run the test again, diluted in half.
-
Be careful not to touch the end of the tube as it
is extremely hot.
Albuminuria
This is the presence of albumin in the urine. Albumin
is a protein, consisting of serum albumin and serum globulin
that has been eliminated from the blood plasma. It contains
carbon, hydrogen, nitrogen, oxygen, and sulfur. Its exact
composition has not been determined.
Urinary albumin does not necessarily indicate
diseased kidneys; it may reflect reactions to toxic and
nontoxic substances originating within the body. Albuminuria is
frequently found in young men who have no other signs of
disease. This condition is usually transitory. However,
albuminuria usually is of clinical significance and generally
requires further examination.
Methods for Measuring Albumin in Urine
The test is accomplished by means of test strips.
Since practically all urine is tested for both glucose and
albumin, the tests are combined in the multitest strips. If
the strips are unavailable, or positive for albumin, the
sulfosalicylic acid method of albumin determination may be
used.
Sulfosalicylic Acid Method of Albumin
Determination
-
If the fresh specimen is clear, the test may be run
without centrifuging. If the specimen is cloudy,
centrifuge about 15 ml at 2,000 rpm for 5 minutes.
-
Pour 2.5 ml of clear urine into a test tube measuring
16 mm x 150 mm.
-
Add 7.5 ml of 3 percent sulfosalicylic acid to the
urine.
-
Mix by inversion and let stand 10 minutes before
reading
-
A white turbidity indicates albuminuria. Compare the
specimen with the standards and report as indicated,
i.e., 1, 5, etc.
CAUTION: The centrifuge is a carefully balanced
machine, and efforts should be made to maintain that
balance. Specimens should be placed directly opposite each
other in the machine. If only one urine specimen is being
centrifuged, place a tube containing an equivalent weight of
water directly opposite the urine.
Microscopic Examination of Urine
Sediment
Usually performed in addition to routine procedures,
this examination requires a degree of skill that can be
acquired only through practice under the immediate supervision
of a competent technician. The specimen should be as fresh as
possible, since red cells and many formed solids tend to
disintegrate upon standing, particularly if the specimen is
warm or alkaline.
Procedure
-
Mix the specimen well.
-
Pour 15 ml of urine into a conical centrifuge tube
and centrifuge at 1,500 rpm for 5 minutes.
-
Invert the centrifuge tube and allow all of the
excess urine to drain out. DO NOT SHAKE THE TUBE WHILE
INVERTED. Enough urine will remain in the tube to
resuspend the sediment. Too much urine remaining will
cause diluting of the sediment and difficulty in
reading.
-
Resuspend the sediment by tapping the bottom of the
tube.
-
With a medicine dropper, mount one drop of the
suspension on a slide and cover it with a coverslip.
-
Place the slide under the microscope and scan with
the low-power objective and subdued lighting.
-
Switch to the high-power objective for detailed
examination of a minimum of 10 to 15 fields.
Clinically Significant Findings
Leukocytes - Normally, 0 to 3 leukocytes
per high-power field will be seen on microscopic
examination. More than 3 cells per high-power field probably
indicates disease somewhere in the urinary tract. Estimate
the number of leukocytes present per high-power field and
report it as the "estimated number per high-power
field."
Erythrocytes - These cells are not usually
present in normal urines. If erythrocytes are found,
estimate their number per high-power field and report it.
Erythrocytes may be differentiated from white cells in
several ways:
-
White cells are larger than red cells.
-
When focusing in with the high-power lens, the red
cells show a distinct circle; the white cells tend to
appear granular and the nucleus may be visible.
-
The addition of one drop of 5 percent acetic acid to
the urine sediment will disintegrate any red cells but
will not affect the white cells except to make the nuclei
more distinct.
Casts - These urinary sediments are formed
by coagulation of albuminous material in the kidney tubules.
They are cylindrical and vary in diameter depending on the
size of the renal tubule or duct of their origin. The sides
are parallel, and the ends are usually rounded. Casts in the
urine always indicate some form of kidney disorder and
should always be reported. If casts are present in large
numbers, the urine is almost sure to be positive for
albumin. Casts containing organized structures are:
-
RBC casts
-
WBC casts
-
Epithelial casts
Types of casts:
-
hyaline
-
waxy
-
granular
-
fatty
Cylindroids - Resemble hyaline casts but
are more slender and have a slender tail that is often
twisted or curled. They frequently are seen along with
hyaline casts and have the same significance.
Other microscopic structures found in urine
are:
These are not generally pathologic unless present
in very large numbers. Certain types of crystals are
pathologic; therefore, all crystals seen should be
reported.
The Hospital Corpsman and Clinical Laboratory
Techniques
As mentioned in the beginning of this chapter, hospital corpsmen
are required to have a basic knowledge of laboratory procedures. It
is not expected that all hospital corpsmen be proficient in all
phases of this field, but it is essential that they know how to
perform the tests mentioned in this chapter, since they are eligible
for duty independent of a medical officer.
The hospital corpsman is not expected to make diagnoses from test
findings or to institute definitive treatment based upon them;
however, with the availability of modern communications facilities,
the results of these tests will greatly assist him or her in giving a
clearer clinical picture to the supporting medical officer.
Needless to say, accuracy, neatness, and attention to detail are
essential to obtain optimum test results. Remember also that these
tests are only aids to diagnosis-many other clinical factors must be
taken into consideration before treatment can be started.
Administrative Responsibilities in the
Laboratory
The ability to perform clinical laboratory tests is a
commendable attribute of the hospital corpsman. However, the
entire effort can come to naught if proper recording and filing
practices are ignored and the test results go astray.
Since the test results are a part of the patient's
clinical picture, their accuracy and veracity are vital. Since
they have a bearing upon the immediate and future medical history,
they are made part of the medical record. Erroneous and inaccurate
laboratory results have been known to cause extensive
embarrassment and medical complications.
As a hospital corpsman, it is your responsibility to
assure effective administration of all laboratory reports in your
department and to make sure that they are properly filed.
Patient Identification
When accepting laboratory requests and specimens,
make absolutely certain that the patient is adequately
identified. Proper identification can prevent a great number of
errors.
Specimen Identification
Make sure that the specimen is in fact that of the
patient submitting it. You need not stand over the patient
while it is being collected; however, keep in mind that there
are instances when it would be advantageous for persons to
substitute specimens.
Use of Proper Forms
The Armed Forces have gone to great lengths to
produce workable and effective laboratory forms to serve their
purpose with a minimum of confusion and chance for error. These
forms are standard forms of the 500 series. Their primary
purpose is to request, report on, and record clinical
laboratory tests. With the exception of the SF-545, Laboratory
Report Sheet, they are multicopy, precarbonized for
convenience. The original eventually is filed in the patient's
clinical record, and the carbon becomes part of the
laboratory's master file. For a complete listing of these forms
and their purposes, refer to chapter 23 of the Manual of the
Medical Department.
Use of Laboratory Forms
It goes without saying that a separate form is used
for each patient and test. The patient's name, rank, Social
Security number, and unit identification will be entered on
each request in sufficient detail to assure proper
identification.
Since the results of the requested laboratory test
are usually closely associated with the patient's health and
treatment, the requesting physician's name and location shall
be clearly stated. This assures that the report will get back
to the physician as expeditiously as possible. There is nothing
more aggravating to both patient and physician than a lost or
misplaced laboratory report.
Since the data requested, the date reported, and the
time of specimen collection are usually important in support of
the clinical picture, these facts should be clearly written on
the request in the areas provided for them.
The type of tests requested should be clearly marked
to eliminate all misunderstanding.
Filing the Laboratory Requests
After the physician has seen the results of the
laboratory tests, the forms must be filed in the clinical
record of inpatients. SF-545, Laboratory Report Sheet, is
provided for this. The originals of the test forms are to be
stapled on this sheet IN CHRONOLOGICAL ORDER and neatly spaced,
as directed on the form. Each sheet will accommodate a certain
number of laboratory reports. DO NOT OVERCROWD with more
reports-use additional sheets if necessary.
The results of the laboratory tests performed on
active duty outpatients will be placed on the SF-545 in the
health (medical) treatment record.
Ethics in the Laboratory
The nature of laboratory tests and their results must be
treated as a confidential matter between the patient, physician,
and the performing technician. It is good practice to prevent
unauthorized access to these reports, to leave interpretation of
the test results to the attending physician, and to refrain from
discussing them with the patient.
References
-
Clinical Diagnosis and Management by Laboratory Methods, ed
16. W. B. Saunders Co, 1979
-
Laboratory Medicine: Hematology, ed 6. C. V. Mosby Co, 1982
-
Hematology for Medical Technologists, ed 5. Lea & Febiger
Co, 1983
-
Hematology: Principles and Procedures, ed 2. Lea & Febiger
Co, 1976
-
Modern Urine Chemistry, Ames Division, Miles Laboratories, Inc
-
A Handbook of Routine Urinalysis, J. P. Lippincott Co,
1983
Approved for public release; Distribution is unlimited.
The listing of any non-Federal product in this CD is not an
endorsement of the product itself, but simply an acknowledgement of the source.
Operational Medicine 2001
Health Care in Military Settings
Bureau of Medicine and Surgery
Department of the Navy
2300 E Street NW
Washington, D.C
20372-5300 |
Operational Medicine
Health Care in Military Settings
CAPT Michael John Hughey, MC, USNR
NAVMED P-5139
January 1, 2001 |
United States Special Operations Command
7701 Tampa Point Blvd.
MacDill AFB, Florida
33621-5323 |
This web version is provided by
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