(Adapted from http://www.biologycorner.com/) A microscope is an instrument that magnifies an object so that it may be seen by the observer. Because cells are usually too small to see with the naked eye, a microscope is an essential tool in the field of biology. In addition to magnification, microscopes also provide resolution, which is the ability to distinguish two nearby objects as separate. A combination of magnification and resolution is necessary to clearly view specimens under the microscope. The light microscope bends a beam of light at the specimen using a series of lenses to provide a clear image of the specimen to the observer. In this lab, parts of the microscope will be reviewed. Students will learn the proper use and care of the microscope and observe samples from pond water.
Your microscope has 4 objective lenses: Scanning (4x), Low (10x), High (40x), and Oil Immersion (100x). In this lab, you will not use the oil immersion lens; it is for viewing microorganisms and requires technical instructions not covered in this procedure. In addition to the objective lenses, the ocular lens (eyepiece) has a magnification. The total magnification is determined by multiplying the magnification of the ocular and objective lenses.
1. Plug your microscope into the power supply and switch on the illuminator. 2. Always start with the stage as low as possible and using scanning objective (4x). Odds are, you will be able to see something on this setting (sometimes it’s only a color). Use the coarse knob to focus: the image may be small at this magnification, but you won't be able to find it on the higher powers without this first step. Move the mechanical stage until your focused image is also centered. 3. Once you've focused using the scanning objective, switch to the low power objective (10x). Use the coarse knob to refocus and move the mechanical stage to re-center your image. Again, if you haven't focused on this level, you will not be able to move to the next level. 4. Now switch to the high power objective (40x). At this point, ONLY use the fine adjustment knob to focus specimens. 5. If the specimen is too light or too dark, try adjusting the diaphragm.
1. Store microscope with the scanning objective in place and the stage in its lowest position. 2. Wrap cords around the microscope. 3. Replace slides to original slide tray.
A large part of the learning process of microscopy is getting used to the orientation of images viewed through the oculars as opposed to with the naked eye. A common mistake is moving the mechanical stage the wrong way to find the specimen. This procedure is merely practice designed to make new users more comfortable with using the microscope.
This portion of the procedure is another practice to demonstrate depth perception. Many new microscope users find it difficult to conceive that the specimen on the slide is in three dimensions. As the stage is moved up and down, different threads will be in focus.
Because it is too hard to find a specimen on high power. Easier to find them and focus them on low power and then increase the magnification and fine-tune the focus to get a better, closer look.
Updated April 17, 2018 By Ellen Murphy
When you change from low power to high power on a microscope, the high-power objective lens moves directly over the specimen, and the low-power objective lens rotates away from the specimen. This change alters the magnification of a specimen, the light intensity, area of the field of view, depth of field, working distance and resolution. The image should remain in focus if the lenses are of high quality.
Changing from low power to high power increases the magnification of a specimen. The amount an image is magnified is equal to the magnification of the ocular lens, or eyepiece, multiplied by the magnification of the objective lens. Usually, the ocular lens has a magnification of 10x. A typical lab-quality standard optical microscope will usually have four objective lenses, running from a low power of 4x to a high power of 100x. With an ocular power of 10x, that gives the standard optical microscope a range of overall magnification from 40x to 1000x.
The light intensity decreases as magnification increases. There is a fixed amount of light per area, and when you increase the magnification of an area, you look at a smaller area. So you see less light, and the image appears dimmer. Image brightness is inversely proportional to the magnification squared. Given a fourfold increase in magnification, the image will be 16 times dimmer.
Going to high power on a microscope decreases the area of the field of view. The field of view is inversely proportional to the magnification of the objective lens. For example, if the diameter of your field of view is 1.78 millimeters under 10x magnification, a 40x objective will be one-fourth as wide, or about 0.45 millimeters. The specimen appears larger with a higher magnification because a smaller area of the object is spread out to cover the field of view of your eye.
The depth of field is a measure of the thickness of a plane of focus. As the magnification increases, the depth of field decreases. At low magnification you might be able to see the entire volume of a paramecium, for example, but when you increase the magnification you may only be able to see one surface of the protozoan.
The working distance is the distance between the specimen and objective lens. The working distance decreases as you increase magnification. The high power objective lens has to be much closer to the specimen than the low-power objective lens in order to focus. Working distance is inversely proportional to magnification.
Microscopes magnify an object's appearance by bending light. Higher magnification means the light is bent more. At a certain point, the light is bent so much that it can't make it through the objective lens. At that point – usually around 100x for standard lab microscopes – you'll need to put a drop of oil between your specimen and the objective lens. The oil "unbends" the light to stretch out the working distance and make it possible to image at high magnifications. |
Why must the specimen be in the Centre of the field of view before you switch from low to high power?
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