The Microscope. Its History, Construction, and Application 15th ed. / Being a familiar introduction to the use of the instrument, and the study of microscopical science

by Jabez Hogg

Method of Employing the Achromatic Condenser to the Greatest Advantage.

Its Illumination.—Good daylight is the best for general work. The microscope should be placed near a window with a northern aspect. Direct sunlight should never be utilised; the best light is that reflected from a white cloud. A good paraffin lamp is the most serviceable artificial source of light, and it is quite under control. As an illuminant more often brought into requisition in the smoky atmosphere of towns, the paraffin lamp is on the whole the handiest and the most useful. If gas-light can be brought into use as suggested for micro-photography, with the incandescent mantle, it will be found to be the purest and best form of artificial illumination for the microscope. Among paraffin lamps those constructed by Baker and Swift are all that can be desired.[28]

Sectional view of the Optical Arrangement of the Aplanatic Bull’s-eye Lens, fitted in gymbal on the front of the lamp.

Fig. 148.—Baker’s Microscope Lamp.

As the chimneys of these lamps are made of metal, and blackened, no reflected light disturbs the eye. Care must be taken to have the wick evenly trimmed; the metal chimney has a glazed front, giving exit to the rays of light, the flat of the flame being used with low powers, and the image of the flame being reflected by a plane mirror to give equal illumination of the whole field. In working with high powers, the lamp is turned with the flame edge-wise, and at the same time the mirror must be dispensed with. By working, as it is termed, directly on the edge of the flame, the illumination is greatly increased, and a band of light can be concentrated on any part of the preparation it is desired to make a careful study of.

To obtain the best results, time and care must be given to the illumination of the object. The lamp and microscope having been placed in position, a low power is first used and the smallest diaphragm. On looking through the microscope it will probably be observed that the image of the diaphragm is not in the centre of the field; by moving the centring screw of the condenser this may be adjusted. The low power is then replaced by a high power, the largest diaphragm used, and the bacteria or diatom brought into focus. The diaphragm must now be replaced by one of medium size, and by racking the condenser up and down, a point will be arrived at when the image of the edge of the flame appears as an intensely bright band of light. If this is not exactly in the centre of the field the centring screw of the condenser must again be adjusted. With regard to the use of diaphragms, various sizes should be tried while focussing with the fine adjustment, at the same time using the correction colour; in this way we obtain the sharpest possible image. When the condenser has been accurately centred, it will still be necessary to focus it for each individual specimen, so as to correct for difference in the thickness of slides and the layers of mounting medium. Correction for different thickness of cover-glasses must be made by the aid of the collar adjustment in the following way: a high-power eye-piece is substituted for the ordinary eye-piece, and the faults in the image will thereby be intensified. By moving the collar completely round, first in one direction and then in the other, while carefully observing the effect of the image, it will be seen to become obviously worse whichever way the collar is turned. The collar must then be turned through gradually diminishing distances until an intermediate point is reached at which the best image results with the high-power eye-piece, and on replacing this by the low-power eye-piece the sharpest possible image will be obtained.

Effect of the Sub-stage Condenser.—The sub-stage condenser gives the most powerful illumination when it has been racked up until it almost touches the specimen. It produces a cone of rays of very short focus, and the apex of the cone should correspond with the particular bacterium or group of bacterias under observation. The effect of the condenser without a diaphragm is to obliterate what Koch has termed a structure picture. If the component parts of a tissue section were colourless and of the same refractive power as the medium in which the section is mounted, nothing would be visible under the microscope. As, however, the cells and their nuclei and the tissues do not differ in this respect, the rays which pass through them are diffracted, and an image of lines and shadows is developed. If in such a tissue there were minute coloured objects, and if it were possible to mount the tissue in a medium of exactly the same refractive power, the tissue being then invisible, the detection of the coloured objects would be much facilitated. This is exactly what is required in dealing with bacteria which has been stained with aniline dyes, and the desired result can be obtained by the use of the sub-stage condenser.

If we use the full aperture of the condenser the greatly converged rays play on the component parts of the tissue, light enters from all sides, the shadows disappear, and the structure picture is lost. If now a diaphragm is inserted, so that we are practically only dealing with parallel rays, the structure picture reappears. As the diaphragm is gradually increased in size the structure picture gradually becomes less and less distinct, while the colour picture, the image of the stained bacteria, becomes more and more intense. When, therefore, bacteria in the living condition and unstained tissues are examined, a diaphragm must be used, and when the attention is to be concentrated upon the stained bacteria in a section or in a cover-glass preparation the diaphragm must be removed and the field flooded with light—(Crookshank).

The wide-angle condenser, it will be understood, consists of a combination of lenses, which concentrate all the light entering them to a small point, and the condenser must be so accurately focussed that this brilliant cone of light, when it emerges from the upper lens of the condenser, falls upon the object from all directions, forming a wide-angle cone of light, at the apex of which the object must be placed (see [Fig. 149]). That is to say, the object is illuminated by a cone of rays passing through it in all directions.

Fig. 149.—Front Lens of Condenser.

There are, however, objects which require a fully illuminated field, when the lamp should be turned round and the Herschel lens condenser (shown in section, [Fig. 148]) should be used to collect the light and throw it upon the mirror. For moderate powers, as a four-tenth or one-fifth, the condenser should be used a little below the focus to give an even illumination over the whole field. Moreover, as to the use of the condenser for defining general objects, it must be borne in mind that to show different kinds of structure different apertures in the iris diaphragm are necessary, and that whereas some objects show their structure better with a large angle of light cut down in intensity by the use of blue glass, others show better with a small pencil of direct rays. For the resolution of diatoms it is often necessary to use oblique light only, and for this purpose diaphragms with central patches are used, the iris diaphragm being opened to its full extent. An annular ring of oblique light emerges from the condenser upon the object, and it is in this manner also that dark-ground illumination is obtained with moderate and low powers.