Anna Lemira Gibson.

Clinical laboratory technic for nurses online

. (page 1 of 12)
Online LibraryAnna Lemira GibsonClinical laboratory technic for nurses → online text (page 1 of 12)
Font size
QR-code for this ebook


GIFT OF

Pacific Coast




fllOLOCY UBRftRY



CLINICAL
LABORATORY
FOR



BY

ANNA L. GIBSON, R.N.

Matron Superintendent of the Collis P. Huntington

Memorial Hospital^ Harvard Medical School^

Boston, Mass.



REVISED EDITION




WHITCOMB & BARROWS

BOSTON, 1922



5




COPYRIGHT 1916 AND 1922

BY
ANNA L. GIBSON



THOMAS TODD Co., PRINTERS
14 Beacon Street, Boston, Massachusetts

MADE IN U S. A.



TO

EMMA M. NICHOLS, R.N.

Formerly Superintendent of Nurses , The Boston City Hospital, Boston, Mass

IN APPRECIATION OF THE
INSPIRATION, KNOWLEDGE, AND TRAINING

WHICH I RECEIVED
FROM ASSOCIATION WITH HER



743540



PREFACE

THIS book owes its existence to the frequent request
on the part of graduate nurses, whom it has been my
pleasure to instruct in laboratory technic during the past
three years, for a simple, comprehensive text-book, that
the nurses might be able to grasp the principles of clinical
laboratory technic.

The book was originally compiled as a handbook for
practical clinical laboratory work, since no single text-
book covered the work.

The arrangement of the several chapters has worked
itself out from a series of lessons which give simple and
reliable methods. By these methods information may be
obtained without unnecessary detail which requires a
considerable knowledge of general chemistry and elab-
orate apparatus.

Standard works on Bacteriology, Chemistry, Hema-
tology, Histology, and Parasitology have been consulted
freely, and references are given at the end of each chapter
from which more detailed information may be obtained.

A great deal of the material and the drawings have
been taken from my notebook.

Grateful acknowledgment is made to Dr. Thomas
Ordway, Dean of Albany Medical College, with whom
I have had the privilege of working the past three
years at the Huntington Hospital ; to Dr. Ellis Kellert,
Director of the Bender Laboratory, Albany, New York,
who gave me valuable assistance the two years we
were associated at the Huntington Hospital ; and to
Dr. Ernest Tyzzer, Assistant Professor of Pathology,



vi Preface

Harvard Medical School, and Director of Huntington
Hospital, Harvard Medical School.

ANNA L. GIBSON, R. N.
BOSTON, MASSACHUSETTS.

September, 1915.

NEW PREFACE

WITH the aim of keeping this book abreast of present-
day laboratory technic, the entire work has been carefully
revised and considerable new matter has been added.
Hospital schools of nursing are being called upon to pro-
duce specialists in many fields of endeavor, and there
should be a broader and more systematic study of the
various branches relating to nursing.

There is no more important subject than clinical labo-
ratory technic, nor one more far-reaching in its influence
on the ultimate success or failure of the work of a nurse.

This book was not written with the thought of mak-
ing nurses diagnosticians, but to help them to have a more
intelligent and more thorough understanding of this sub-
ject in its relation to nursing.

I have had the advantage, as formerly, of the help and
advice of professors in Harvard Medical School. I am
especially indebted to Dr. Robert B. Greenough, Assistant
Professor of Surgery at Harvard Medical School and
Director of the Cancer Commission of Harvard Univer-
sity, for encouragement in my teaching of this subject;
to Dr. Francis Peabody and Dr. George Minot, Profes-
sors of Medicine, Harvard Medical School, for instruc-
tion in laboratory technic ; also to various authors whose
works I have consulted, and to publishers who have so
willingly granted me permission to use illustrations from
their books. ANNA L. GIBSON, R. N.

1922.



CONTENTS

CHAPTER PAGE

I. LABORATORY EQUIPMENT . . . . . . i

II. THE MICROSCOPE . . . . . . 16

III URINE . . . . - .'- . 20

IV. FECES . '. . 56

V. GASTRIC CONTENTS 70

VI. SPUTUM . . .. . . i v . . .84

VII. THE BLOOD . . . . . . . . 90

VIII. BACTERIA . 141

IX. CULTURE MEDIA . 166

X. BODY FLUIDS . . 180

XI. MILK 187

XII. PREPARATION OF TISSUE . . . , . 192

APPENDIX 197



vii



CHAPTER I
LABORATORY



Glassware { ^ *\*,, a rl J >0 '

It is very important that all glassware used in the
laboratory should be made of Bohemian or Jena glass.
Many test tubes are made of the ordinary glass, which is
silicate of calcium and sodium, and sometimes lead oxide
is used instead of calcium carbonate.

This glass is easily acted upon by chemical substances,
and should not be subjected to heat.

Tubes, flasks, and beakers should be made of potassium
carbonate glass Bohemian glass as this glass is char-
acterized by its great hardness, difficult fusibility, and by
its resistance to the action of chemical substances, sudden
changes of temperature, and high steam pressure.

The best glass tubing and rods are made of Jena
glass, as it is chemically superior to the Bohemian glass,
more resistant to acidulous fluids and sudden changes of
temperature.

Care of Glassware

All glassware used in the laboratory work must be
thoroughly clean before using. New glassware should
be placed in 0.5% solution of nitric acid to remove the
alkali frequently present; thoroughly rinse in running
water.

Glass slides are cleaned by immersing in cleaning solu-
tion, then washing in water; dry with a towel and flare



2 t Clinical Laboratory Technic

both sides over a Bunsen flame. Oil which has dried on
slides can be removed with xylol.

Old test tubes containing culture media should be re-
sterilized for one hour, or boiled for one hour in a 5 %
solution of soda; this destroys the bacteria and loosens
the material is? -the tubes.

Test tubes 'anil' flasks are dried in the autoclave, then
plugged with non-absorbent cotton and sterilized one-half
hour at 15 pounds pressure.

A good cleaning fluid is made as follows :

Potassium bichromate 60 c.c.

Concentrated sulphuric acid 300 c.c.

Water 400 c.c.

Dissolve the potassium bichromate in water with heat.

Cool, then add slowly the sulphuric acid.

Glass Droppers and Capillary Pipettes

Take a piece of tubing and heat it in the middle of a
Bunsen flame, revolving the tubing while heating; and
when it becomes soft in the center, remove from the flame
and with a steady pull separate the ends. Cool, file, and
break off. Flare the rough ends in the flame.

Glass Stirring Rods

Take a piece of glass rod, file off the desired length,
then round off the rough ends in the flame by constant
rotation.

Weights and Measures

The Analytical Balance. The poise in the ordinary
balance is not disturbed by slight variations of weight,
but in chemical analysis a more sensitive instrument is



Laboratory Equipment



necessary. The beam is made as light as possible, the
bearings sharp and hard, the adjustments capable of being
brought to the last degree of . refinement and provided
with appliances for arresting its action at will. It is in-
closed in a glass case for protection against dust, moisture,
and currents of air.

The beam is divided by notches into tenths, and carries
weights shaped as rid-
ers, and these riders
lessen in value as they
are moved towards the
center.

A rider weighing
.01 gram in the pan
weighs .09 gram at first
notch from the pan,
.08 gram at the second.

Large brass weights
equal grams; large
platinum weights, 0.5 gram ; small weights, .05 gram ;
and the rider, .01 gram.

The gram is the unit of weight and equals the weight
of i c.c. of distilled water at 4 C.

I kilogram = 1,000 grams = 100,000 centigrams =
1,000,000 milligrams.

I kilogram = 2.20462 pounds = 35.2739 ounces
15,432.35 grains.

Always lift the weights with the forceps provided.

A watch glass is used as a receptacle for reagents
weighed. First ascertain the weight of the glass and add
the amount to the required amount of the reagent.

Note. Objects to be weighed should be placed on the left-hand
pan, and the weights on the right-hand pan.




ANALYTICAL BALANCE



4



Clinical Laboratory Technic



A meter equals 39.37 inches.

A cubic meter is the unit of space for the number of
organisms in air. It contains 1,000 liters. It is equal
to 1.308 cubic yards or 35.316 cubic feet. 1,000 cubic feet,
the unit of space in disinfection, is equal to 28.3 plus cubic
meters.

A cubic centimeter is the unit of space for organisms



a
sq.m.m.





c.c



sq.inch.



METRIC ''SYSTEM 2

INCHES n 21 31

I I I I I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I 1 I I I I I I I I I I I I I I I I I I J J I I 1

COMPARISON OF INCHES AND CENTIMETERS

in water, milk, vaccines, etc. There are approximately
16 drops in i c.c.

Cubic millimeter is the unit of space for blood cells.
There are 1,000 cubic millimeters in i cubic centimeter,
and 1,000,000 cubic millimeters in i liter.

A liter is the unit of space for volumetric solutions.
It contains 1,000 cubic centimeters, and is equal to 1.0567
quarts or 33.8 ounces. A liter of distilled water equals
I kilogram.



Laboratory Equipment



Decimal Table




Length


1,000


kilometer


100


hectometer


10


decameter


I


Meter


O.I


decimeter


0.0 1


centimeter


O.OOI


millimeter



The arc is the unit of surface and is the square of ten
meters.



Weight Capacity

kilogram kiloliter

hectogram hectoliter

decagram decaliter

Gram Liter

decigram deciliter

centigram centiliter

milligram milliliter or cubic centimeter



Reagents

The reagent bottles should be made of Jena glass, which
is free from lead and other impurities, and these bottles
should be fitted with ground glass stoppers. All reagents
should be chemically pure (C. P.)-

Liquid Reagents

Nitric acid, C. P. (HNO 3 ) ; acetic acid, (HC 2 H 3 O 2 ) ;
sulphuric acid, C. P. (H 2 SO 4 ) ; hydrochloric acid, C. P.
(HC1) ; ammonic hydrate, (NH 4 OH) ; sodic hydrate,
(NaOH),U. S. P.

Solid Reagents

Cupric sulphate, caustic soda, sodium chloride, potas-
sium iodide, potassium chromate, ammonium sulphate,
magnesium sulphate, ammonium chloride, sodium acetate,
potassium ferrocyanide, potassium acetate, guaiac, ben-
zidin, potassium chlorate, picric acid, citric acid, lead
acetate, sulphanilic acid, sodium nitrite, sodium carbon-
ate, mercuric chloride, potassium bromide, sodium nitro-
prusside, alphol naphthol, phenylhydrazin hydrochlorate,



6 Clinical Laboratory Technic

di-methyl-amino-azo-benzine, di-methyl-paraphenylene-di-
amine, Eosin Gruber, w. g.

The different alcohols used in making up the various
reagents are: absolute alcohol, which contains not less
than 99% by weight of pure ethyl alcohol, C 2 H 5 OH ;
alcohol of a stated percentage, e. g., 50%, means a mix-
ture with water which contains the stated percentage,
e - > 5%> by volume of pure ethyl alcohol; methyl
alcohol, a pure substance, CH 3 OH, prepared by the puri-
fication of commercial wood spirit ; and methylated alcohol
or methylated spirit, which may be used instead of pure
ethyl alcohol in preparing solutions of various percent-
ages of alcohol. Methylated alcohol is a mixture of
19 parts of ethyl alcohol and i part commercial methyl
alcohol. Commercial methyl alcohol is impure and must
not be used in making Eosin-Azur, Louis Jenner's, Leish-
man's, Wright's, or Romanowsky stains.

The amounts of distilled water and absolute alcohol
required to produce saturated solutions of dyes in common
use are indicated in the following table :

Alcohol (ex.)

7

2-5

7

I
I

7
10

To Remove Glass Stoppers

If there is any difficulty in removing the glass stoppers
from the reagent bottles, they are easily loosened by
gently tapping the neck of the bottle with another piece
of glass.





Dye


Water


Bismarck Brown


I


7


Fuchsin (Basic)


I


10


Gentian Violet


I


7


Hematoxylin


I


2


Methyl Violet


I


5


Methylene Blue


I


7


Thionin Blue


I


5



Laboratory Equipment
Table of Equivalents



Liquids


Approximate


Accurate


i minim


= 0.06


c.c.


0.06 1


c.c.


16 "


== i.


c.c.






i fl. dram
i fl. ounce


= 4-
= 30.


c.c.
c.c.


3.697
29-574


c.c.
c.c.


i pint
i gallon


= 500.
== 4,000.


c.c.
c.c.


473-197
3,785.


c.c.
c.c.


Solids










i kilogram


= 2.2


Ibs. av.






I gram


= is-


grains


15432


grains


I milligram


= A


grain


0.015.


\ grain



Solutions

Approximately correct solutions are made by using the
following method :

For i -i, ocx) (TO%) use T S grains to a liter.
For i-ioo (i%) use 5 grains to the ounce.
EXAMPLES, i. Make 2,500 c.c. of a 1-500 solution of
potassium permanganate. 15 grains, or i c.c., to 500 c.c.
is a 1-500 solution. If 15 grains, or i c.c., is used to every
500 c.c., for 2,500 c.c. we would use as many c.c. as there
are 500 in 2,500, or 5 c.c. (75 grains).

2. Make 500 c.c. of i -10,000 solution caustic potash.
If it takes 15 grains to 1,000 c.c. to make a 1-1,000 solu-
tion, to make a 1-10,000 solution, which is 10 times weaker,
take -^Q of 15 grains, or 1.5 grains, for 1,000 c.c. of a
1-10,000 solution, and for 500 c.c. of a i-io,ooc solution
take ^ of 15 grains, or 0.75 grain.

3. Make 75 c.c. of a \% solution of acetic acid. \%
equals 1-300. 75 c.c. -f- 300 c.c. = .25 c.c. If i c.c. is
used to every 300 c.c., to 75 c.c. we would use \ as much ;
therefore with a pipette measure 0.25 c.c.



8 Clinical Laboratory Technic

Saturated Solution

The solubility of many metallic salts, acids, alkalies,
sugars, and organic products is very great, yet there is
a limit beyond which it is not possible to dissolve a solid
in liquid. This limit is the point of saturation.

Concentrated Solution

When we speak of a concentrated solution we mean
the amount of the solute in a given quantity of the
solvent.

Standard Solution

A standard solution is one whose concentration is
known. This can be made of any strength. To obtain
a standard solution use a solution of sodium hydroxide
of known strength. This gives a standard of hydrogen
fixing power. Units of this solution will be equivalent
to definite amounts of acid in the solution neutralized.

;*

Normal Solution

A normal solution is a standard solution which con-
tains in one liter the hydrogen equivalent in grams of
the active reagent. A normal volumetric solution is
made by dissolving the hydrogen equivalent or atomic
weight divided by its valence in distilled water and
making its volume up to one liter. The molecular weight
in grams of a base, salt or acid, is divided by the valence,
and the valence of a base is the number of hydroxyls
combined with it ; the valence of an acid is the number
of replaceable hydrogen atoms which it contains.

If the molecule of the element is univalent, one liter
will contain the weight in grams equal to the molecular
weight of the element; if bivalent, a weight in grams



Laboratory Equipment 9

equal to one-half its molecular weight ; if trivalent, a
weight equal to one-third its molecular weight.

Common Elements



(1920) Symbols


Atomic


Symbols


Atomic






Weights






Weights


Aluminum


Al


27.1


Gold


Au


197.2


Antimony


Sb


120.2


Hydrogen


H


1.008


Arsenic


As


74.96


Iodine


I


126.92


Barium


Ba


137.37


Iron


Fe


55.84


Bismuth


Bi


208.


Lead


Pb


207.20


Boron


B


10.09


Magnesium


Mg


24.32


Bromine


Br


79.92


Mercury


Hg


200.6


Cadmium


Cd


II2.4


Nitrogen


N


14.008


Calcium


Ca


40.07


Oxygen





16.


Carbon


C


12.005


Phosphorus


P


31.04


Chlorine


Cl


3546


Potassium


K


39.10


Chromium


Cr


S 2 -


Silver


Ag


107.88


Cobalt


Co


58.97


Sodium


Na


23.00


Copper


Cu


63.57


Sulphur


S


32.06


Fluorine


F


10.









EXAMPLE. NaOH is univalent. Na = 23, O = 16,
H = 1.008. The sum of these atomic weights equals
40.008. Dissolve 40.008 grams of NaOH in distilled
water and make up to one liter.

H 2 SO 4 is bivalent. II 2 = 2.016, S = 31.83, O 4 =
63.52. The sum of these atomic weights equals 97.366.
As it has two replaceable H atoms, it would contain in
one liter of water one-half this number, or 48.683 grams
of absolute H 2 SO 4 .

Solutions of this strength are designated by the capital
letter N ; of twice this strength, by 2N ; one-half or one-
tenth, by N/2 or N/io. Equal volumes of normal solu-
tions react together completely. One liter of normal
NaOH or KOH will neutralize one liter of normal
H 2 SO 4 or HNO 3 .



io Clinical Laboratory Technic

METHOD. Clean volumetric flasks and beakers with
cleaning solution; rinse thoroughly in running water,
twice with distilled water, and then with a small portion
of the solution they are to contain.

Pick out pure crystals of oxalic acid, 1 place in a watch
glass, and weigh with analytical balances, the weight of
the glass having been previously ascertained.

The molecular weight of oxalic acid is 126.048. As it
is diabasic, divide by two. 63.024 grams are necessary to
make one liter of N solution. To make N/io, take 6.3024
grams.

Pour the crystals into a dry beaker, rinsing off the
watch glass with distilled water ; stir with a glass rod
until dissolved.

Place the glass rod in the neck of a liter volumetric
flask and carefully pour the oxalic solution from the
beaker. Rinse the beaker several times with distilled
water, then wash off the rod and fill the flask from a wash
bottle of distilled water up to the mark; the last few
drops can be blown from the wash bottle.

As the inside neck of the flask is wet from the addition
of water, a filter paper which has been rolled up is in-
serted and rotated until the neck is dry, being careful not
to touch the fluid.

Stopper and thoroughly mix.

Having N/io oxalic acid, N/io sodium hydroxide is
prepared as follows: Sodium hydroxide is very hygro-
scopic, therefore a normal solution cannot be accurately
prepared by weight. Weigh out 5 grams and dissolve in
1,100 c.c. of water. Titrate with the oxalic solution.

If accurately measured should give N/io solution in which the
error is less than i %.



Laboratory Equipment



n





B



A. Graduated burette and support.

B. Graduated pipette.

C. Volumetric flask.

D. Erlenmeyer flask.



Indicators are substances which assume a deep color
in the presence of certain other substances, or change
sharply from one deep color to another.

EXAMPLES. Phenolphthalein, a very feeble acid, is
colorless in the presence of acids (i.e., hydrion), and red
in the presence of alkalies (i.e., hydroxidion). Litmus
is red with acids, and blue with alkalies. The change of
color depends upon a chemical interaction.



12 Clinical Laboratory Technic

Titration

A definite amount of oxalic acid solution is measured
with a graduated pipette and placed in a flask. Add
50 c.c. of distilled water, then 3 drops of alizarin or
phenolphthalein, as an indicator that the point of neutral-
ization or end of the reaction can be accurately deter-
mined. Phenolphthalein is used when neither ammonia
nor bicarbonates are titrated. In titrating acid and alkali,
always run the alkali into the acid.

EXAMPLE. Pipette 10 c.c. of the oxalic acid solution
into an Erlenmeyer flask; do not blow out the amount
left in the end of the pipette. Pour 50 c.c. of distilled
water into the flask and add 3 drops of phenolphthalein.
Rinse out a burette with distilled water and with the
sodium hydroxide solution, then fill with the solution
until it rises above the zero mark. A few c.c. are run out
until the mark is reached. Place the flask of oxalic solu-
tion on a filter paper for a white background and run in
the sodium hydroxide solution, a few c.c. at a time, shak-
ing the flask gently. Add the sodium solution until there
is a distinct pinkish color, which is the end of the reaction.

If 9.8 c.c. of the sodium hydroxide solution were re-
quired to produce a distinct pink color, it is stronger than
the N/io oxalic acid solution, as only 10 c.c. would have
been necessary if the sodium solution had been N/io.
Therefore 9.8 c.c. of the sodium solution are equivalent
to 10 c.c. of the N/io oxalic solution. The sodium hy-
droxide must be diluted in the proportion of 9.8 to 10.
Measure exactly 1,000 c.c. of the too concentrated sodium
hydroxide solution and add 20 c.c. of distilled water.



Laboratory Equipment



Equivalent Fahrenheit and Centigrade Tables



AUTOCLAVE TEMPERATURE

Sterile dressings, media, disin-
fection of spore-bearing bac-
terial contamination



C.

125
1 20

H5
no

105

100



F.
258
250
240
230

220

212



FREEZING TEMPERATURE

Preserving biological products ;
post-mortem material

C. F.



40



4
3

2

I




BODY TEMPERATURE

Growth of important patho-
genic organisms

C. F.



40

39
38
37



104



98.2



ROOM TEMPERATURE

Culturing gelatin

(melting point, 25 C.)

as in water work



C.

14
23

22
21
20



F.

75



69.8
68



PARAFFIN AND PASTEURIZING

Bacterial vaccines and
paraffin bath

C. F.

80 176

75

70 158

65

60 140

Fahrenheit scale is divided into equal divisions or
degrees, the lowest of which is a mixture of equal parts
of sal-ammoniac and snow, and the highest, the boiling



14 Clinical Laboratory Te clinic

point of pure water; the freezing point of water on this

scale is 32.

Centigrade scale is divided into 100 equal parts, or
degrees, the space of expansion from the
freezing point to the boiling point of pure



joo_:



aio-

water. The number of degrees between
the boiling point and freezing point in Centi-
grade is 100, and in Fahrenheit it is 212 32
or 180. EXAMPLE. 100:1 80:: degree to be
g converted :X. By division with 20, 5:9::
degree to be converted :X; i.e., the degree
g to be converted is multiplied with 9, the re-
g suit divided with 5, and 32 added to the
result.



H

u



p Centigrade X

-^^ - \- 32 = Fahrenheit.

< . c '



I Fahrenheit 32 X 5 ^ ,. -,
g _^ ^ == Centigrade.

es Q



Laboratory Rules

1. All possible cleanliness should be ob-
j_ served in the care of apparatus.

2. The hands should be washed with a
disinfectant after working with pathogenic
bacteria, and then with soap and water.

3. Pencils and labels should never be moistened with
the lips.

4. Discarded cultures should be covered with a disin-
fectant, then resterilized.

5. Culture media should be put in receptacles provided
for that purpose, and not in the sink.



Laboratory Equipment 15

6. Pipettes which have been used to handle infectious
material should be placed in a glass receptacle containing
cleaning fluid.

7. Any infectious material dropped on the table or floor
should be immediately wiped with a disinfectant.

8. All bottles should be plainly labeled.

9. All cultures and tissue specimens should be labeled
with the patient's name, source, and date.

10. Sterilize the platinum loop before and after use.

11. When using the autoclave, see that there is suffi-
cient water before turning on the pressure.

12. Whenever material is placed in the centrifuge
tubes, see that they balance evenly.



CHAPTER II



THE MICROSCOPE

One of the most important pieces of apparatus used in
the laboratory is the microscope.

i. The microscope consists of a tube 160 millimeters
(6.4 in.) long, having two systems
of lenses, which conduct the rays as
they pass from the objective to the
oculars.

2. The Ocular, or eyepiece, is at
the upper end of the tube. There are
various oculars and they are num-
bered from one to ten, the magnify-
ing power of the ocular increasing
as the number advances.

3. The Objective is a system of
converging lenses at the lower end
of the tube, which forms a magnified
inverted image of the object.

4. The Stage, with clips to hold a
slide in position during examination.

5. The Reflector or small mirror has two sides, a con-
cave and a plane mirror. The reflector directs the rays
of light upward through the object in the optical axis of
the microscope.

6. The Sub-Stage Condenser is a system of lenses
between the stage and the mirror. These lenses collect
and condense the rays coming from the reflector so that

16




MICROSCOPE



The Microscope 17

they are focused upon the object, thus giving a brilliant
illumination.

7. The Iris Diaphragm controls the intensity of the
illumination and is just below the sub-stage condenser.
The gradations of light are obtained by means of a small
lever.

8. The Coarse Adjustment is a rack and pinion mech-
anism which rapidly raises and lowers the barrel and its


1 3 4 5 6 7 8 9 10 11 12

Online LibraryAnna Lemira GibsonClinical laboratory technic for nurses → online text (page 1 of 12)