Elementary technique in histology and bacteriology

UNIVERSITY OF CALIFORNIA
AT LOS ANGELES

GIFT OF CAPT. AND MRS.
PAUL MCBRIDE PERIGORD

ELEMENTARY TECHNIQUE

IN

HISTOLOGY AND BACTERIOLOGY

BY

ERNEST B. HOAG, A. B., B. S.

Instructor in Zoology and Physiology,

THROOP POLYTECHNIC INSTITUTE,

Pasadena, Cal.

H. KAHN, PHAR. M.

Assistant Demonstrator in Bacteriology,

NORTHWESTERN UNIVERSITY MEDICAL SCHOOL,

Chicago.

CHICAGO:

THE W. T. KEENER CO.
1896.

136516

Entered according to Act of Congress, in the year 1895, by

Ernest B. Hoag and H. Kahn,
in the Office of the Librarian of Congress, at Washington.

5.5-. 5-

PREFACE.

This guide in elementary technique is intended for
the use of medical and other students beginning
work in either Histology or Bacteriology.

ERRATA.

Page 12. 19, :23, for "creasote" read "creosote."

Page 15, line 3, for "tissues" read "tissue."

Page 40, for "Shafer" read -'Sehafer."

Page 16, 17, 20 for "flxitive" read "fixative."

Page 71, eighth line from bottom, for "or covered" read "are covered."

For "liquifying" read ''liquefying."

are usuany taxen ror granted, ana wmcn tneretore, often"
greatly perplex the student.

It is not expected that the book will furnish the ex-
perienced student with much information; it is written
to help the beginner and while it is thought that it will
be sufficiently complete for the requirements of all such,
we especially hope that it will serve as an introduction
to more advanced works, and that the result will be,
a clear understanding of the elementary methods, and
therefore a much more intelligent use of the advanced
books on Histology and Bacteriology.

It will be noted, that no attempt has been made to
introduce descriptive Histology and scarcely any de-
scriptive Bacteriology, but that the book is confined to
Technique.

S5". 5"

PREFACE.

This guide in elementary technique is intended for
the use of medical and other students beginning
work in either Histology or Bacteriology.

In its main features it is a new arrangement
of what is found in other books on these subjects, and
in many instances direct quotations are made.

The absence of any concise, and yet sufficiently full,
account of methods which will enable the beginner to
form from the first, clear ideas of the various steps is
the only apology offered for adding another book to the
many excellent ones now to be found.

It is hoped that this little book will answer that com -
mon question of the beginner, "What shall I do next:"'
We have tried to make plain those simple points which
are usually taken for granted, and which therefore, often
greatly perplex the student.

It is not expected that the book will furnish the ex-
perienced student with much information; it is written
to help the beginner and while it is thought that it will
be sufficiently complete for the requirements of all such,
we especially hope that it will serve as an introduction
to more advanced works, and that the result will be,
a clear understanding of the elementary methods, and
therefore a much more intelligent use of the advanced
books on Histology and Bacteriology.

It will be noted, that no attempt has been made to
introduce descriptive Histology and scarcely any de-
scriptive Bacteriology, but that the book is confined to
Technique.

PREFACE.

We are particularly indebted for valuable assistance
received from Dr. Stanley P. Black, Pathologist at
Mercy Hospital. Dr. F. X. Walls, and Dr. E.G. Conklin
of Northwestern University, Dr. A. C. Eychleshymer
of Chicago University, and Dr Adolph Gehrmann of
College of Physicians and Surgeons, Chicago, have also
rendered us much assistance.

Among the books consulted, we have drawn more
particularly for Part I from Lee's Vade Mecum, Stir-
ling's Histology, Schafer's Histology, Von Kahlden's
Pathological Histology, and Lehrbuch der Histologie
und Mikroskopischen Techink. (Bohm und Davidoff),
and for Part II from Migula, Sternberg, Schenk, Novy,
and Praenkel. Suggestions and criticisms from those
interested in the material here presented, will be greatly
appreciated.

September, 1895.

PART I.

HISTOLOGY.

HISTOLOGY AND BACTERIOLOGY. 5

2nd. Hardening the elements of a tissue so that their
structure will remain as nearly normal in appearance as
possible, after the various reagents for their preparation
have been used.

The fixing of a tissue is of the greatest importance,
and final success or failure with the sections depends
very largely upon this step.

One must know what fixing agent to use for a par-
ticular tissue; how long to allow it to act; with what to
wash out the fluid, and what strength of alcohol to use
after washing.

Secure perfectly fresh tissue if possible. Use
small pieces, and 15 to 20 times their volume of the
fixing fluid.

There are many good fixing agents and the choice of
one depends upon the kind of tissue and the result de-
sired. For most purposes the following will be found
satisfactory fixing fluids, and among them, corrosive
sublimate, absolute alchohol, Flemming's fluid and Per-
enyi's fluid are particularly recommended.

FIXING AGENTS IN GENERAL USE.

1. PERENYI'S FLUID.— Formula on page 32.

Objects are left in the fluid from 3 to 5 hours for small
embryos and 4 to 12 hours for the tissues of vertebrates,
and then transferred directly to 70-75 per cent, alcohol
for at least 24 hours. They may then be placed in 80
per cent alcohol and left in this until wanted, or they
may be dehydrated at once by carrying them through
the alcohols, including absolute. This is a very valua-
ble fluid for both embryonic and adult tissues. No seri-
ous results follow when a tissue is left in the fluid for a
number of hours. Borax carmine may be added to 70
per cent alcohol so that the hardening and staining is
done at the same time.

HISTOLOGY AND BACTERIOLOGY. 5

2nd. Hardening the elements of a tissue so that their
structure will remain as nearly normal in appearance as
possible, after the various reagents for their preparation
have been used.

The fixing of a tissue is of the greatest importance,
and final success or failure with the sections depends
very largely upon this step.

One must know what fixing agent to use for a par-
ticular tissue; how long to allow it to act; with what to
wash out the fluid, and what strength of alcohol to use
after washing.

Secure perfectly fresh tissue if possible. Use
small pieces, and 15 to 20 times their volume of the
fixing fluid.

There are many good fixing agents and the choice of
one depends upon the kind of tissue and the result de-
sired. For most purposes the following will be found
satisfactory fixing fluids, and among them, corrosive
sublimate, absolute alchohol, Flemming's fluid and Per-
enyi's fluid are particularly recommended.

FIXING AGENTS IN GENERAL USE.

1. PERENYI'S FLUID. — Formula on page 32.

Objects are left in the fluid from 3 to 5 hours for small
embryos and 4 to 12 hours for the tissues of vertebrates,
and then transferred directly to 70-75 per cent, alcohol
for at least 24 hours. They may then be placed in 80
per cent alcohol and left in this until wanted, or they
may be dehydrated at once by carrying them through
the alcohols, including absolute. This is a very valua-
ble fluid for both embryonic and adult tissues. No seri-
ous results follow when a tissue is left in the fluid for a
number of hours. Borax carmine may be added to 70
per cent alcohol so that the hardening and staining is
done at the same time.