The Microscope

Owen G. Sunga, MD Ryan Barrow 2008

Owen G. Sunga, MD Ryan Barrow 2008

Objectives
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Know the Basics

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Parts and Functions Use Types and Differences

Owen G. Sunga, MD Ryan Barrow 2008

Historical Timeline
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1609 Galileo Galilei develops a compound microscope with a convex and a concave les. 1665 Robert Hooke publishes Micrographia and coins the word cell for structures he discovers in cork bark 1674 Anton van Leeuwenhoek improves on a simple microscope for viewing biological specimens 1931 Ernst Ruska starts to build the first electron microscope

Owen G. Sunga, MD Ryan Barrow 2008

Types of Microscopes

Owen G. Sunga, MD Ryan Barrow 2008

Types of Microscopes I
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Compound Microscope
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Light microscope that has two converging lens systems: the objective and the eyepiece

Owen G. Sunga, MD Ryan Barrow 2008

Types of Microscopes II
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Electron Microscope
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Any class of microscopes that use electrons instead of light to form images of very small objects such as individual parts of small living things

Owen G. Sunga, MD Ryan Barrow 2008

Types of Microscopes III

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Transmission Electron Microscope

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Microscope used to reveal the internal structure of microbial and cell types. The electron beam is scanned, or swept over a metal coating to form a three-dimensional image. Can produce three-dimensional images of individual molecule and atoms, as well as movies

Scanning Electron Microscope

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Scanning Tunneling Microscope

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Owen G. Sunga, MD Ryan Barrow 2008

The Compound Microscope

Owen G. Sunga, MD Ryan Barrow 2008

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Owen G. Sunga, MD Ryan Barrow 2008

The Compound Microscope

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Base
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Supporting structure that generally contains the source Converges light beams to pass through the specimen Controls the amount of light passing through the specimen

Condenser
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Iris Diaphragm
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Owen G. Sunga, MD Ryan Barrow 2008

Compound Microscope II
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Objective Lens
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Magnifies image Conveys light to the ocular lens Magnifies the image from the objective. A microscope with one ocular lens (eyepiece) is monocular; a microscope with two ocular lenses is binocular

Body Tube (Arm)

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Ocular Lens (Eye piece)

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Owen G. Sunga, MD Ryan Barrow 2008

Compound Microscope III

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Mechanical Stage
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Allows precise control in moving the slide Knob used to locate specimen Knob used to bring the specimen into sharp focus

Coarse Adjustment
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Fine Adjustment
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Owen G. Sunga, MD Ryan Barrow 2008

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Owen G. Sunga, MD Ryan Barrow 2008

Compound Microscope IV
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Total magnification
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Calculated by multiplying the magnifying power of the objective lens by the magnifying power of the ocular lens. Increased magnification is no value unless good resolution can also be maintained.

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Owen G. Sunga, MD Ryan Barrow 2008

Properties of Light

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Owen G. Sunga, MD Ryan Barrow 2008

Properties of Light
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Reflection
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Light strikes an object and bounces back Passage of light through an object
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Transmission
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Must be one of these 2 for a microscope

Absorption
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Taken up by that object and used as energy

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Photosynthesis

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Owen G. Sunga, MD Ryan Barrow 2008

Properties of Light II
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Refraction
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Bending of light as it passes through one medium to another of different density

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Owen G. Sunga, MD Ryan Barrow 2008

Properties of Light II
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Diffraction
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Occurs when light rays are bent as they pass through a small opening, such as a hole, a slit, a space between two adjacent cellular structures, or a small, high powered, magnifying lens in a microscope. The bent light rays distort the image obtained and limit the usefulness of the light microscope.

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Owen G. Sunga, MD Ryan Barrow 2008

Immersion Oil
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Immersion Oil
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Same index of refraction as glass Used to replace air to prevent refraction at a glass-air interface Works as a secondary lens Allows a clearer and larger image

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Owen G. Sunga, MD Ryan Barrow 2008

Proper Procedures
1.

2.

Carry the microscope with one hand under the base while grasping the arm with the other hand Place microscope ~5 inches from the edge of the table

Be careful of the cord when moving around the lab tables!!

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Owen G. Sunga, MD Ryan Barrow 2008

Proper Procedures
3. Rotate the nosepiece to obtain the proper objective lens Make sure the lens clicks into place!! 4. Look through the eyepiece and switch on the lamp This is your field of view!

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Owen G. Sunga, MD Ryan Barrow 2008

Proper Procedures
5. Place slide on stage, center specimen over stage 6. Locate specimen on low power using coarse adjustment 7. Carefully switch to medium or high power 8. Focus image with fine adjustment

Avoid damaging slide!

Do not use course adjustment with high power!
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Storing the Microscope

Four steps prepare the microscope for storage:
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the 10X objective is in place

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the stage is all the way down the power is off the cord is wrapped around the base

3. 4.

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Owen G. Sunga, MD Ryan Barrow 2008