Clinical laboratory methods; a manual of technique and morphology designed for the use of students and practitioners of medicine

Front matter

flass l?S5 7

Book Mj-

CopightN

Preface

Xo attempt has been made to include within the present volume a multiplicity of methods; in fact, the aim of author has been to select one method or more of proved value. Nor have the more exact, time-consuming methods of physiological chemistry been drawn upon; in his daily work the average clinician has not the time, if he has the ability, to employ them.

Free use has been made of the following works: Emerson's " Clinical Diagnosis." Wood's "Chemical and Microscopical Diagnosis," Simon's "Clinical Diagnosis," Sahli's ■'Klinische Untersuehiingsinethoden. " Hoppe-Seyler's "Handbueh der chemischen Analyse." Hammarsten's "'LHkrbueh der physiologischen Chemie." Xeubauer-Huppert 's "Analyse des Haras." Schmidt and Strasburger's "Die Faezes des Mensehen." Braun's '"Thierische Parasiten des Menschen." Blanc-hard's -'Traite de Zoologie Medieale." Cabot's "Clinical Pathology of the Blood." Xaegeli's "Blutkrankheiten und Blutdiagnostik. " and Turk's "Vorlesungen ueber klinische Haematologie." Other authors have been consulted less freely. To the more recent literature direct reference has been given in the form of footnotes ; in all instances the writer has endeavored to give proper credit to authors.

It is hoped that the volume will prove helpful to medical students who have completed a course in Clinical Pathology and to practitioners of medicine, or that it may serve as a supplement to a course of laboratory lectures.

For the original illustrations in black and white the author is indebted to Dr. James S. Brotherhood. It is a pleasure, also. to acknowledge his indebtedness to his wife and to his mother for assistance in the preparation of the index and in other ways.

Roger Sylvester Morris.

St. Louis.

Chapter I The Urine Page

Collection of the Urine ....... 1

Preservation of the Urine 2

Macroscopic Examination of the Urine .... 3

Color of Urine 4

Quantity of Urine . . . 4

Reaction of Urine 4

Quantitative determination of urinary acidity: Folin's

method . 5

Specific Gravity 6

Hiifner's hypobromite method ...... 8

Uric Acid 9

Qualitative determination of uric acid .... 9 Quantitative determination of uric acid: Method of

Folin and Shaffer 10

Quantitative determination of ammonia: Folin's method — the vacuum distillation method — the formalin titration method — Shaffer's modification of Schlosing's

method 13-19

Nitrogen 20

Kjeldahl's method for determination of total nitrogen . 20

Chlorids 23

Qualitative test . 23

Quantitative determination of chlorids: Harvey's modification of Volhard's method 24

ix x CONTENTS

PAGE

Sulphates 26

Obermayer's test . .27

Jaffe's test ■ 27

Qualitative tests: Heat and acetic acid test — Heat and nitric acid test — Heller's test — Potassium ferrocyanid and acetic acid test ....... 28-34

Quantitative determination of albumin : Tsuchiya 's modification of the Esbach method — Removal of albumin from the urine ........ 35-36

Bence- Jones' Body 36

Albumose • 37

Qualitative tests: Trommer's test — Fehling's test — Almen-Nylander 's test — The fermentation test — Cippolina's modification of the phenylhydrazin test . .38-42 Quantitative estimation of glucose: Benedict's first method — Benedict's second method — Polariscopic determination — Robert's specific gravity method — Measurement of carbon dioxid gas formed during fermentation 44-51

Levulose 53

Seliwanoff's test, as modified by Borchardt ... 53

The phenylhydrazin test 55

Maltose . . 56

Lactose 56

Saccharose 57

Pentose 57

The phloroglucin test 57

The orcin test ......... 58

Bial's modification of the orcin test .... 58

Glycuronic Acid 59

B. Tollens' test 60

Alkaptonuria 61

Qualitative tests: Gunning's test — Lieben's test — Legal's

test — Lange's test 62-64

Diacetic Acid 65

Gerhardt's test .65

CONTENTS xi

PAGE

Diacetic Acid (Continued)

ft Oxybutyric Acid 67

Black's test 67

Hart's test 68

Urobilinogen 70

Ehrlich's alclehyd test 70

Spectroscopic determination ...... 71

Schlesinger 's test 71

Jaffe's test 72

Bile Pigments 73

Qualitative tests: Foam test — Gmelin's test — Rosenbach's modification of Gmelin's test — Huppert's test — Hammarsten's test ....... .73-75

Hematoporphyrin . . . . . . . . .76

Spectroscopic determination . . . . 78

The guiac test ......... 81

Heller's test 82

Teichmann's hemin-crystal test ..... 82

The Diazo Reaction 83

Chyluria 84

Lipuria . . . 86

Ferments in the Urine ....... 86

Wohlgemuth 's method for the determination of diastase 87

Determination of lipase according to Hewlett . . 88

The unorganized sediments : The quadriurates of sodium and potassium — Uric acid — Calcium oxalate — Calcium sulphate— Monocalcium phosphate — Hippuric acid — Cholesterin — Xanthin — Hematoidin — Tyrosin — Leucin — Cystin — Tricalcium and trimagnesium phosphates — Calcium carbonate — Ammoniomagnesium phosphate — Ammonium biurate — Neutral magnesium phosphate — Neutral calcium phosphate ..... 91-99

Xll

CONTENTS

The Urinary Sediments (Continued)

The organized sediments : Epithelial cells — Pus — Blood — Casts — Mucous threads — ' ' Clap threads ' ' — Gonococcus — Treponema pallidum — Bacillus tuberculosis

Animal Parasites in Urinary Passages Trichomonas vaginalis Filaria bancrofti

Dioctophyrae renale .

Schistosoma hematobium Prostatic Fluid Functional Dl\gnosis of the Kidney

The phthalein test of Rovntree and Geraghty

Chapter II The Gasteic Juice

Test Breakfasts 125

Examination of the Fasting Stomach . . . .127 Macroscopic Examination of the Gastric Contents . . 128

Quantity 128

Odor 128

Mucus 128

Color 129

Food . 129

Chemical Examination of the Gastric Contents . '. 131

Hydrochloric acid: Qualitative tests for free hydrochloric acid (von den Yelden's methyl violet test: Giinzberg's test; Tropeolin test; Congo-paper test: Topfer's test; Sahli's desmoid test) — Quantitative determination of gastric acidity (Topfer's method for free hydrochloric acid; Other indicators; Titration of total acidity) — The

hydrochloric acid deficit 131-138

Lactic acid: Qualitative tests for lactic acid (Ueffelmann's test; Strauss' test; Kelling's test) . . 141-112 Butyric acid 112

CONTENTS xiii

Chemical Examination of the Gastric Contents (Continued)

PAGE

Acetic acid 143

Pepsin: Qualitative test for pepsin — Quantitative methods (Mette's method as modified by Nierenstein and

Eennin: Qualitative test for rennin .... 145

Eennin zymogen ........ 146

Pathological enzyme in the gastric contents . . . 146

Mucus . 148

Microscopic Examination of the Gastric Contents . . 149

CHAPTER III THE FECES Macroscopic Examination of the Feces . . . . 152

Form 153

Color .153

Mucus 154

Gallstones 155

Parasites 155

Intestinal Test Diet 155

Diet No. I 156

Diet No. II 156

Weight of Dried Feces 157

Chemical Examination of the Feces 157

Urobilin (Hydrobilirubin) : Schmidt's test — Schlesin-

ger's test — Spectroscopic determination .... 158

Bilirubin: Schmidt's test — Gmelin's test . . 158-159 Blood: Weber's test — The guiac test — Teichmann's

hemin crystal test 159-161

Trypsin: Method of Gross for the determination of

XIV

CONTEXTS

PAGE

Chemical Examination of the Feces (Continued)

Amylase: Wohlgemuth 's method for the determination

of amylase, as modified by Hawk ..... 163

Microscopic Examination of the Feces .... 166 Food remnants ........ 167

Bacteria 168

Cells 169

Crystals 170

Intestinal parasites: Protozoa, Rhizopoda — Flagellata — Infusoria — Nematodes — Trematodes — Cestodes . 172-194

Preservation of gross specimens of cestodes and other parasites : Permanent preparations of flat worms (Method of Mink and Ebling — Boggs' method — Creosote

Accidental contaminations ...... 203

CHAPTER IV THE SPUTUM Amount

.

.

Character

Odor

Consistence .

.

Air Bubbles .

.

Dittrich's Plugs

.

Bronchial Casts

.

Ccrschm ann's Spirals

.

Layer Formation .

.

.

Examination of fresh spu turn

.

Yellow elastic tissue

.

Curschmann 's spirals

.

Alveolar epithelial cells

Dust cells

' ' Heart-failure sells'

CONTENTS xv

PAGE

Microscopic Examination {Continued)

Red blood corpuscles 210

Pus cells 211

Eosinophilic leukocytes , 211

Charcot-Leyden crystals . . . . . . .211

Microorganisms in sputa: Bacillus tuberculosis (Ziehl- Neelsen method; Antiformin method for the detection of tubercle bacilli) — Diplococcus pneumoniae (Gram's method of staining; Welch's capsule stain) — Bacillus influenzae — Bacillus diphtherias (Neisser's staining method; Beall's method) — Actinomyces bovis — Streptothrix ep-

pengeri — Blastomycetes 212-223

Animal parasites in the sputum: Entamoeba histolytica — Entamoeba tetragena — Trichomonads — Paragonimus westermanii — Echinococcus cyst .... 224-225

CHAPTER V Obtaining Blood for Examination 226

Blood Stickers 226

The hemocytometer ........ 227

Procedure in counting the erythrocytes : Diluting fluids — Filling the pipette — Filling the counting chamber — The enumeration of the cells — Calculation of the result —

Cleaning the apparatus 229-234

Counting the leukocytes: Diluting fluid — Filling the pipette — Filling the counting chamber — The enumeration of the leukocytes — Calculation of the result — Biirker's modification of the Thoma counting chamber . 236-239 Counting the eosinophilic leukocytes: Dunger's method 241 Counting the blood platelets: Method of Wright and Kinnicutt 242

XVI

CONTENTS

PAGE

Tallqvist's method . 244

Sahli's hemometer 245

The Fleischl-Miescher hemoglobinometer . . . 249 Haldane's hemoglobinometer ...... 252

Dare's hemoglobinometer 252

Sulphhemoglobinemia 252

Methemoglobinemia 252

Color Index 253

Volume Index . . 253

Measuring the Diameter of Cells 255

Viscosity of the Blood and Other Fluids . . . 256

Method of Hess 256

The Specific Gravity of the Blood 259

The Coagulation Time of the Blood 260

Method of Brodie and Russell, as modified by Boggs . 260 Milian's method, as modified by Hinman and Sladen . 263 The Resistance of the Red Blood Corpuscles . . . 264 The Examination of Fresh and Stained Preparations of

Blood 265

The cleaning of cover glasses and slides .... 265 Examination of the fresh blood : Sealing the fresh specimen — The preparation of dry (permanent) blood smears

— Fixation of blood smears 265-271

Staining the blood: Vital staining of the blood — Vaughan's method; Method of Widal, Abrami, and Brule ; The ' ' dry ' ' method of vital staining ; The staining of dried blood films — Methylene blue ; Eosin ; Hematoxylin; Carbol-thionin. Staining mixtures of two or more stains — Ehrlich 's triacid stain ; The Romanowsky stains (Wilson's stain, Leishman's stain, Giemsa's stain) ; Jenner's stain; Methyl green-pyronin mixture of Pappenheim; The iodin reaction of the leukocytes

273-295 Differential counting of the leukocytes: Normal leukocytes — Pathological leukocytes .... 296-299

CONTENTS xvii

PAGE

The Examination of Fresh and Stained Preparations op Blood (Continued) The normal and pathological red blood corpuscles: Erythrocytes — Erythroblasts — Abnormalities in the staining of the red corpuscles .... 301-302 Demonstration of protozoa in the blood: Malarial parasites . 305

Examination of the blood for animal parasites: Filaria bancrofti — Trichinella spiralis ...» 308-310

CHAPTER VI Specific Gravity 312

Albumin Content 312

Incoagulable Nitrogen 313

A Protein Precipitable in the Cold by Dilute Acetic

Acid 315

Cytology 315

Cerebrospinal Fluid 318

Cells of the cerebrospinal fluid 318

Method of counting the cells 318

Bacteriology of the cerebrospinal fluid .... 319

Globulin content: Method of Noguchi — Method of Ross and Jones ........ 319-321

The Wassermann reaction in cerebrospinal fluid . . 321

Index 323

LIST OF ILLUSTRATIONS

FIG. PAGE

1. — Apparatus for the quantitative determination of ammonia according to Folin 14

2. — Apparatus for the determination of ammonia according

to Shaffer . . . 16

3. — Digesting rack for the Kjeldahl nitrogen determination 21 4. — Distilling apparatus for the Kjeldahl nitrogen determination 22

5. — Lohnstein's areometer ....... 51

6. — Lohnstein's fermentation saccharometer for undiluted

7. — Absorption spectra 79

8. — The Sydenham sedimenting glass 89

9. — Treponema pallidum; Spirochaeta refringens . . 112

10. — Embryo of Filaria bancrofti ...... 117

11. — Ova of Dioctophyme renale ...... 118

12. — The Autenrieth-Konigsberger colorimeter as modified by Rowntree and Geraghty for the determination of

phenolsulphonephthalein ...... 123

13. — Parasitic amebaa 173

14. — Trichomonas intestinalis 177

15. — Lamblia intestinalis 178

16. — Balantidium coli 179

17. — Ovum of Necator americanus . . . . . .. 181

18. — Necator americanus . . . . . . . 185

19. — The rhabditiform embryo of Strongyloides stercoralis . 186

20. — Ovum of Strongyloides stercoralis 187

21. — The rhabditiform embryo of Strongyloides stercoralis

and the embryo of the hookworm .... 188

22. — Ovum of Oxyuris vermicularis 188

23. — Ovum of Trichuris trichiura 189 xix xx LIST OF ILLUSTRATIONS

FIG. PAGE

24. — Ovum of Ascaris lumbricoides : tlie same under high

focus, showing the albuminous coating . . . 190

25. — Unfertilized ovum of Ascaris lumbricoides . . . 191

26. — Ovum of Schistosoma haematobium .... 193

27. — Ovum of Schistosoma japonicnm 193

28. — Gravid proglottis of Tenia saginata : ovum of Tenia

saginata ; gravid proglottis of Tenia solium . . 195

29. — Ovum of Dibothriocephalus latus 196

30. — Gravid proglottis of Dibothriocephalus latus . . . 197

31. — Ovum of Hynienolepis nana ...... 197

32. — Ovum of Hynienolepis diminuta ..... 198

33. — Dipylidium caninum. showing an egg capsule and a

free ovum 199

34. — Tyroglyphus siro. the cheese-mite and ovum . . . 2'~>2 35. — Actinomyces hominis. showing club-shaped extremities

to the rays 222

36. — Blastomycetes in sputum 223

37. — Ovum of Paragonimus westermanii from the sputum . 224

38. — Sediment from echinococcus cyst 225

39. — The Thoma-Zeiss hemocytometer ..... 227

■40. — The Xeubauer ruling of the hemocytometer . . . 22 S

41. — Biirker's hemocytometer 239

42.— The Sahli hemometer 245

43. — The Fleischl-Miescher hemoglobinometer . . .250 44. — The viscosimeter of Hess . . . . . .257

45. — Boggs' modification of the coagulometer of Brodie and

Russell 261

46. — Diagram to illustrate the movement of the cells during

coagulation 262

CLINICAL LABORATORY METHODS