How Does a Compound Light Microscope Work?

By: Spencer Lambert March 14, 2013

This document will serve to show the different parts of a compound light microscope, and how they work to magnify specimens. The document will be focused towards high school seniors or college freshmen who are being introduced to light microscopy. The audience should have a basic understanding of biology and simple light microscopes. They should also understand the principles of magnification and illumination. It will serve as a guide to help them understand the instrument they are using.

The invention of the light microscope is credited to spectacle-makers Hans and Zacharias Janssen, who first invented the device in the 1590s. The light microscope, however, was not greatly popular until it was popularized by Anton van Leeuwenhoek in the early 1700s. Van Leeuwenhoek was the first person to view bacteria, yeast, and blood circulation. Another great achievement in light microscopy was the description of eukaryotic cells by Robert Hooke, who sectioned plant cork tissue and viewed it under a light microscope. These early achievements were, however, achieved with relatively primitive single lens microscopes. Modern compound microscopes use multiple lenses to collect light and a separate set of lenses to focus the light into the eye. These microscopes feature more

reliable light sources and lenses of greater magnification and clarity than earlier single lens microscopes, letting the most complex light microscopes resolve objects up to 0.275 microns in size.

The following section outlines the parts of a modern compound light microscope, and how they work together to make the device work. They will be presented by following the light path from light source to eyepiece. Please refer to Image 1 for a visual of the parts

Image 1. Parts of a compound light microscope labeled with light path

a.
While many different light sources may be used in a compound light microscope, the most common light source is a lamp backed by a reflector that reflects light through the collector lens. Most microscopes will have an intensity knob to adjust the light emitted from the light source to allow for optimal clarity of the subject.

b.

The mirror reflects the light emitted by the light source upwards at a 90 degree angle into the field diaphragm. It allows for a more compact design in modern compound light microscopes with the light source housed within the base of the microscope.

c.
The field diaphragm controls the diameter of the light beam that is emitted onto the sample. It allows the user to control how much of the specimen is viewable, and a fine tune of the intensity of the light

d.
The field lens serves to focus light towards the condenser. It is not found on all light microscopes.

e.
The condenser is a special lens designed to focus light from the field lens or diaphragm on the sample. The condenser is fully adjustable, which allows the user to view different layers of thin samples by manipulating the condenser. The condenser can be moved up or down, but the default position is up.

f.
The mechanical stage is the part of the microscope that houses the sample. It will usually contain a clip or vise to hold the specimen in place, and, in higher end models, can be moved horizontally by an adjustment knob on the microscope. When loading a sample onto the stage, the stage should always be completely down, and the lowest objective should always be used first when raising the stage.

g.
The course and fine focusing knobs are used to focus the light coming from the sample into the objective. These knobs physically move the stage up and down. The course adjustment knob, the wider of the two knobs, should only be used to bring the sample into view on the lowest objective. Use of the course knob on higher objectives could cause collision between the sample and objective and potential damage to the microscope. The fine adjustment knob is used to fine tune the focus of the sample on any magnification.

h.
The revolving nosepiece is a rotating structure that houses the microscopes multiple objective lenses. Light microscopes typically have between three and four objective lenses, depending on whether or not it has an oil immersion lens. The objectives are a number of single or multielement compound lenses housed in a cylindrical structure. They focus to collect light from the sample and to focus it into a real image in the tube of the microscope. It is important to never touch the actual objectives, but to use the revolving nosepiece to move from objective to objective. The oil immersion lens should never be used without oil. It can severely damage the microscope.

i.
A microscope with a trinocular head has three potential exits for the light coming through the objectives. Two exits go to the binocular eyepiece for viewing, and a third allows the light to exit into a camera adapter at the top of the microscope. The camera adapter allows for the use of digital photography to photograph samples. You will most likely not use any digital photography in any entry level courses requiring light microscopy.

j.
The eyepiece houses the ocular lens. In the case of your microscope, the eyepiece contains two ocular lenses, allowing for viewing of the specimen with both eyes. The ocular lens bring the real image produced by the objectives into focus for the human eye. The top piece of the left side of the eyepiece can be turned to allow users with glasses to focus on samples without wearing their glasses.

k.
The stand and arm make up the body of the microscope. When moving the microscope, you should always grasp it by the arm and stand, in an upright position. The ocular lenses are not attached to the eyepiece and can fall out of the microscope is not held properly.

The light microscope has been a fixture of the biological sciences for over 400 years, and while compound light microscopes use the same principles as simple microscopes, they are able to achieve much greater clarity and magnification. Modern compound light microscopes have features such as the condenser and multiple objectives that allow them to resolve extremely small objects. These technological breakthroughs have helped students and researchers alike view wonders that could never be seen by the naked eye.

http://www.leica-microsystems.com/uploads/tx_templavoila/light-microscopes-940x310.jpg http://zeiss-campus.magnet.fsu.edu/tutorials/basics/fielddiaphragm/index.html (image 1modified from this image) http://www.microbehunter.com/2008/12/31/parts-of-a-compound-microscope/

http://www.sonlight.com/uploads/9sm-course-and-fine-focus-k.jpg (image modified from this image)

http://resource.rockyview.ab.ca/rvlc/science8/Microscope%20Objective%20Lenses.jpg