on the eyepiece the total magnification of the microscope is obtained. By the magnification of the objective is meant the ratio of the distance of distinct vision to the focal length of the objective. As powerful achromatic objectives show differences of chromatic magnification in the same way as apochromats, compensation eyepieces can be used in combination with these objectives.
Illuminating Systems
Most microscopic observations are made with transmitted light; an illuminating arrangement is therefore necessary, and as the plane of the object is nearly always horizontal or only slightly inclined, the illuminating rays must be directed along the optical axis of the microscope.
To fully utilize the aperture of the system all dispersing rays in the object-space of the objective must be retained in the image-space of the illuminating system. When this occurs the greatest brightness will be obtained if the corresponding diaphragms of the two systems coincide; ie. the field-diaphragm on the image-side of the observing system with object-side of the illuminating system, and the exit pupil of the illuminating system with the entrance pupil of the objective.
For slight magnifications a revolving plane mirror fixed below the object for altering the direction of the rays suffices. For this mirror to illuminate all the points of the objective so that the rays fill up the objective it must not be too small, and should be as near as possible to the stage plate, and the source of light must be considerably extended (fig. 40). Diffused daylight is very suitable. If the aperture of the objective is increased, the diameter of the illuminating surface must also be increased so that the system is quite filled up, from which it follows that this method of illuminating soon fails. The possibilities of illuminating with a concave mirror seem a little more favourable. As a rule a concave mirror of similar aperture is fitted on the other side of the plane mirror. With the concave mirror an image of the source of light can be thrown upon the object. The distance of the concave mirror from the stage plate is about equal to its focal length. This is also the most suitable distance when diffused daylight is used, but it is too short with artificial light; the distance between the stage plate and the mirror should then be increased, so that an image of the source of light can be thrown upon the object. It is simpler to place an illuminating lens in front of the source of light so that the source falls approximately at the front focus of this lens and consequently is represented at infinity through the illuminating lens. By a correct choice of the focal length of the illuminating lens in relation to the focal length of the mirror, it is possible to choose the size of the image of the source of light so that the whole object-field is uniformly lighted.
Too much light is useless for observing delicately coloured or colourless preparations, whose parts only become visible as a result of slight differences of diffraction. Then it is necessary to use powerfully concentrated cones of light. The apparatus must be such that the apertures of the illuminating rays can easily be altered, e.g. by inserting diaphragms in the course of the rays of the illuminating cone below the stage plate (fig. 40, PP). This concentration is most easily produced by sliding or revolving diaphragms. A series of holes of different sizes perforate a revolving disk below the stage plate at an equal radial distance from the axis of the disk, so that the holes can be brought under the preparation in turn, the centre of the diaphragms always being a continuation of the optical axis of the microscope.
Fig. 41.—Cylinder Diaphragm (Zeiss). | |
| Fig. 40.—Mirror Illumination. | |
| M1＝plane-, M2＝curved mirror. | |
| O＝object; L1＝front lens of microscope; | |
| PP＝diaphragm. |
The so-called cylinder diaphragms (fig. 41) are used especially in German microscopes. A changeable diaphragm is placed at the upper end of a short tube which can be moved in a case below the stage in the direction of the optical axis. By bringing the diaphragm nearer the object the aperture of the rays is increased; if the diaphragm is removed farther from the object the cone of rays is diminished (cf. fig. 40). These diaphragms are sometimes fitted in a slide, so that it is possible to move the diaphragm sideways and give oblique illumination (see below).
With very powerful objectives these methods are insufficient; and a condenser is fitted below the stage plate. As a rule an iris diaphragm, which can be moved sideways, is now fitted below this condenser; below is the mirror which can be moved in all directions. The Abbe apparatus consists of a condenser, movable iris diaphragm, and mirror (fig. 42).
Fig. 42.—Abbe Illuminating Apparatus with Ordinary Condenser (Zeiss).
The whole apparatus can be focused by a rack and the button s. The iris diaphragm can be regulated by the lever p; it can also be turned to one side round the pivot z, so that the condenser k can be removed or changed. The correct direction can be given to the illuminating cone by the mirror m. It is often desirable to pass from direct to oblique lighting. The Abbe apparatus makes this easy. The iris diaphragm i is pushed to the side by the rack and pinion t n. The chief cone of rays then enters obliquely into the objective, the angle between the direct cone of rays and the diffraction spectrum of the first order can then become as large again as with direct lighting, and still be taken up in the objective. Oblique lighting, however, can only be in an azimuth, so that the object must be turned in order that the details may be observed. Hence a condenser, for lighting with very oblique cones, must have about the same aperture as the objective, and therefore be of very wide aperture; they therefore closely resemble microscope objectives in construction. Especially powerful achromatic condensers are really only magnified microscope objectives, with the difference that they are not corrected for the thickness of the cover slip, but for the thickness of the glass on which the object is placed. For exceptionally accurate work microscope objectives are sometimes used as condenser systems. When using immersion objectives, an immersion condenser must also be used if rays of extreme obliquity are wanted, for, in consequence of the total reflections, rays can only come from the upper plane surface of the condenser, which have not a larger inclination to the axis than about 41°, varying according to the refractive index of the glass. In order to let highly inclined rays pass out from the condenser, some immersion liquid must be placed between the upper surface of the condenser and the object slide. Condensers are for this reason also constructed with apertures up to 1·40.
Vertical Illuminators.—Opaque objects can only be seen by reflected light. With low magnifying systems and a large free object distance, ordinary good daylight is sufficient. If the objects have a low reflecting power, or if a slightly higher magnification is needed, the lighting can be improved by optical system.
To examine small opaque objects with a high magnification the Lieberkühn mirror, so named after its inventor, was formerly much used. This was a concave mirror, pierced in the middle, fixed
to the objective, and directed towards the object and with such a