say, five
minutes.

Fix, stain, etc., wet, as under (a), and examine.

(B) ~Fixed.--2.~ In ~stained preparations~.

(a) To determine points in _morphology_:

_Shape_ (_vide_ classification, page 131).

_Size_:

(a) Prepare cover-slip film preparations at the various ages, and fix
by exposure to a temperature of 115 deg. C. for twenty minutes in hot-air
oven.

(b) Stain the preparations by Gram's method (if applicable) or with
dilute carbol-fuchsin, and mount in the usual way.

(c) Measure (_vide_ page 66) some twenty-five individuals in each film
by means of the Ramsden's or the stage micrometer and average the
result.

_Pleomorphism_; If noted, record--

The predominant character of the variant forms.
On what medium or media they are observed.
At what period of development.

(b) To demonstrate details of _structure_:

_Flagella_: If noted, record--

Method of staining (_vide_ page 101).
Position and arrangement (_vide_ page 136).
Number.

_Spores_: If noted, record--

Method of staining.
Shape.
Size.
Position within the parent cell.
Condition, as to shape, of the parent cell (_vide_
page 139).
Optimum medium and temperature.
Age of cultivation.
Conditions of environment as to temperature,
atmosphere.
Method of germination (_vide_ page 140).

_Involution Forms_: If noted, record--

Method of staining.
Character (e. g., if living or dead).
Shape.
On what medium they are observed.
Age of medium.
Environment.

_Metachromatic Granules_: If noted, record--

Method of staining.
Character of granules.
Number of granules.
Colour of granules.

~3. Staining Reactions.~--

1. _Gram's Method._--Positive or negative.

2. _Neisser's Method._--If granules are noted, record--

1. Position.
2. Number.

3. _Ziehl-Neelsen's Method._--Acid-fast or decolourised.

4. _Simple Aniline Dyes._--(Noting those giving the best results, with
details of staining processes.)

Methylene-blue }
Fuchsin } and their modifications.
Gentian violet }
Thionine blue }

BY BIOCHEMICAL METHODS.

Test cultivations of the organism for the presence of--

Soluble enzymes--proteolytic, diastatic, invertase.

Organic acids--(a) quantitatively--i. e., estimate the total acid
production; (b) qualitatively for formic, acetic, propionic, butyric,
lactic.

Ammonia.

Neutr