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PREFACE.

This book is designed especially for practical use in pathological laboratories, both as a guide to beginners and as a source of reference for the advanced. We believe that the book will also meet the wants of practitioners who have more or less opportunity to do general pathological work.

Every autopsy presents for solution a problem which may be simple or complex. The known quantities are certain clinical symptoms and physical signs; the unknown quantities are not only the gross and microscopic lesions that may or may not have given rise to the symptoms and signs, but also the etiology of the lesions and the order of their sequence. The solution of the problem often requires the highest skill in post-mortem, bacteriological, and histological technique, but in its solution lies the fascination of pathological work.

It has seemed advisable to us to present, so far as possible, a consecutive statement of the methods employed in solving the various problems that arise, so as to avoid the repetitions that necessarily occur when the three usual divisions of the subject are separately considered by different writers. It is hoped that this method of presenting the subject will bring the student to the realization that the mechanical performance of a post-mortem examination and the inspection of the gross lesions constitute usually only the beginning of the solution of the problem, which should be investigated

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bacteriologically, histologically, and chemically as far as our present knowledge will permit.

We should particularly advise the routine bacteriological and histological examination of the more important organs in all suitable cases. Naturally, the autopsies in which the lesions are due to a single etiological factor are the most valuable and instructive for a clear understanding of the pathological processes present.

Besides the methods of post-mortem examinations and of bacteriological and histological investigations connected with autopsies, we have added the special methods employed in clinical bacteriology and pathology.

In the parts devoted to Bacteriology and to Pathological Histology we have not endeavored to make an exhaustive collection of methods and formulæ, but rather to select those which have been found of the greatest service in practical work.

To Dr. A. H. Wentworth, Assistant in Children's Diseases in the Harvard University Medical School, we are indebted for the sections on the Blood and on Malaria, and for the notes in regard to Lumbar Puncture.

Boston, August, 1897.

IV. THE METHODS OF STUDYING BACTERIA IN CULTURES.

1. Cover-glass Preparations from Cultures, 98.—The Staining of

Spores, 100.-.

- - The Staining of Flagella, 100: Löffler's Method, 102; Pitfield's

Method, 103; Bunge's Method, 103; Van Ermengem's Method, 104.-2.

Methods of Obtaining Pure Cultures, 105.—Method of Isolation of a

Bacterium in Pure Culture from a Mixed Growth, 107: The Plate Method of

Petri, 110; Esmarch's Method of Roll-cultures, 110.–The Determination of

the Motility of Bacteria, 111.-3. The Inoculation of Animals, 112.-

Guinea-pigs, 112.—Rabbits, 113.—Mice, 115.— The Care of Animals, 117.-

4. Cultivation without Oxygen (Anaërobic Cultures): Method of

Liborius, 117; Method of Buchner, 119; Esmarch's Method, 119; Bouillon

Cultures under Hydrogen, 119.

V. BACTERIOLOGICAL DIAGNOSIS.

Staphylococcus Pyogenes Aureus, 121.-

:- Staphylococcus Pyogenes Albus and

Citreus, 124.-Staphylococcus Epidermidis Albus, 124.—Staphylococcus Cereus

Albus and Flavus, 124.–Streptococcus Pyogenes, 124.—Pneumococcus, 128. —

Gonococcus, 130: Special Culture-media, 131; Serum Agar-agar, 131 ; Urine-

Serum-Agar-agar, 132.—Micrococcus Tetragenus, 133.—Diplococcus Intracel.

lularis Meningitidis, 135.—Bacillus Diphtheriæ, 137.- Bacillus of Typhoid

Fever, 141: Differential Diagnosis between the Bacillus of Typhoid Fever and

the Bacillus Coli Communis, 143.—Bacillus Coli Communis, 145.—Bacillus

Tuberculosis, 148.-Spirillum of Asiatic Cholera (Comma Bacillus), 152.—

Bacillus of Anthrax, 156.-Bacillus Pyocyaneus (Bacillus of Green Pus), 160.

-Bacillus of Influenza, 162.-- Bacillus of Glanders (Bacillus Mallei), 164.—

Bacillus Proteus (Proteus Vulgaris), 167.—Bacillus Mucosus Capsulatus, 168.-

Bacillus of Tetanus, 171.- Bacillus Aērogenes Capsulatus, 173.—Bacillus of

Malignant Edema, 175.-Actinomyces, 176.

Introduction, 204.–Laboratory Outfit: Microscopes, 204.-Freezing Micro-

tome, 206.—Celloidin Microtome, 209.—Paraffin Microtome, 210.—Paraffin

Bath, 210.–Vulcanized Fiber, 201.-Knives, 212.-Running Water, 213.-

Slides, 213.-Cover-slips, 214.–Staining Dishes, 214.—Metal Iustruments, 215.

- Bottles, 216.-Examination of Fresh Material, 216.-Indifferent Fluids,

217.—Macerating Fluids, 218.-Examination of Fluids, 218.-Injections,

218.—Cold Injection-masses, 219.–Warm Injection-masses, 219.--Fixing Re-

agents, 220.--Alcohol, 222.—Zenker's Fluid, 223.-Orth's Fluid, 224.-

Flemming's Solution, 224.—Hermann's Solution, 225.—Pianese's Solution,

225.—Rabl's Chromo-formic Acid Solution, 225.-Corrosive Sublimate, 225.—

Formaldehyde, 226.—Boiling, 227.-Müller's Fluid, 227.-Marchi's Fluid, 228.

-Erlicki's Fluid, 228.—Decalcification, 228.—Directions for Using Nitric

Acid, 229.—Phloroglucin and Nitric Acid, 229.—Picric Acid, 230.- Trichlor-

acetic Acid, 230.-Imbedding Processes, 230.—Celloidin, 231.—Imbedding

in Celloidin, 231.—Imbedding in Paraffin, 234.—Serial Sections by the Celloidin

Method, 237.--Serial Sections by the Paraffin Method, 239.--Staining Solu-

tions : Hematoxylin and Hematein Stains, 239.-Carmine Stains, 244.--Aniline

Dyes, 245.—Diffuse Stains, 249.—Combination Stains, 250.— Pianese's Staining

Solutions and Staining Methods, 250.—Orcein, 253.- Iodin, 253.—Lugol's Solu-

tion, 254.-Acid Alcohol, 254.—Aniline Water, 254.-Carbolic-acid Water, 255.

-Mayer's Glycerin-albumin Mixture, 255.—Clearing Reagents, 255.— Mount-

ing Reagents, 258.-Metallic Stains or Impregnations, 258.—Silver, 258.

Gold, 260.-Osmic Acid, 261.-Staining Methods, 262.-Nuclear Stains,

265.—Alum-hematoxylin Stains, 266.— Aqueous Alum-hematoxylin; Delafield's

Hematoxylin; Ehrlich's Acid Hematoxylin, 267.—Mayer's Hemalum, 268.-

Heidenhain's Hematoxylin Stain, 268.--Carmine Stains, 268.—Aniline Dyes as

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