Microscopes Orlando Science Center.

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Presentation transcript:

Microscopes Orlando Science Center

What is a microscope Microscopes are instruments designed to produce magnified visual or photographic images of small objects. The microscope must accomplish three tasks: produce a magnified image of the specimen separate the details in the image render the details visible to the human eye or camera Orlando Science Center

Visualizing Cells

The history of the microscope Light Microscopes have been used for four centuries. The word was first used for magnifying instruments in 1625 Many people experimented with making microscopes The Greeks & Romans used “lenses” to magnify objects over 1000 years ago. Hans and Zacharias Janssen of Holland in the 1590’s created the “first” compound microscope Light microscopes can now magnify up to 1000x The “First” Microscope Zacharias Jansen 1588-1631 Orlando Science Center

History of the Microscope Antoine van Leeuwenhoek 1st to see single-celled organisms in pond water He called them “wee beasties”

History of the Microscope 1655 – Robert Hooke used a compound microscope to observe pores in cork He called them “cells” because they reminded him of the monastery cubicles where the monks lived

The history of the microscope Anthony van Leeuwenhoek and Robert Hooke made improvements by working on the lenses Anthony van Leeuwenhoek 1632-1723 Robert Hooke 1635-1703 Robert Hooke published his microscope drawings in a book called Micrographia Orlando Science Center

The history of the microscope Orlando Science Center

A light microscope is also known as an optical microscope Types of Microscopes Light (Compound) Microscope First type of microscope Most widely used Use reflected or transmitted light which is detected by the eye, or sometimes a camera. Light passes through lenses to produce image Lower magnifying power. Can magnify up to 2000x A light microscope is also known as an optical microscope Orlando Science Center

Types of Microscopes Stereoscopic Microscope Gives a three dimensional view of an object. (Examples: insects and leaves) Used for dissections Used for viewing whole specimens Orlando Science Center

Types of Microscopes Confocal Laser Scanning Microscope (CLSM) A laser beam is used to illuminate spots on the specimen A computer compiles images created from each point to generate a 3-dimensional image Used on specimens that are too thick for a light microscope Orlando Science Center

Types of Microscopes Electron Microscope A beam of electrons passes through lenses to produce image Detectors sense the interactions which create images of the sample High magnifying power Used to observe VERY small objects: viruses, DNA, parts of cells. Orlando Science Center

Introduction to Electron Microscopy Electron Microscopes were not developed until the Twentieth Century The first Transmission Electron Microscope (TEM) was built in 1932 An electron beam is directed at a very thin slice of specimen stained with metal ions The heavily stained material absorbs electrons, but the lightly stained material allows the electrons to pass through These electrons strike a fluorescent screen and create an image Can magnify up to 250,000x Transmission Electron Microscope

Introduction to Electron Microscopy The first Scanning Electron Microscope (SEM) was built in 1942 The electron beam is focused on a specimen coated with a very thin layer of metal The electrons that bounce off the specimen form an image on a fluorescent screen The SEM shows three dimensional images of the surface Can magnify up to 100,000x

Electron Micrographs Scanning electron microscope (SEM) Transmission electron microscope (TEM) Figure 4.1B

Electron Micrographs Scanning electron microscope (SEM) Transmission electron microscope (TEM) Figure 4.1B

Introduction to Electron Microscopy The Scanning-Tunneling Microscope was developed in 1982. A needle-like probe is used to measure differences in voltage caused by electrons that leak, or tunnel, from the surface of the object being viewed A computer tracks the movement of the probe The computer generates a three-dimensional image of the specimen’s surface Objects as small as individual atoms can be viewed Can be used to study living organisms

Light vs Electrons Electrons can be focussed using magnetic lenses Shorter wavelength than visible light

The advantages of using electrons Large depth of field More of the sample is in focus at one time Higher resolution Smaller features can be imaged Analysis The electron beam interacts with the sample enabling information on composition to be collected using additional detectors

Parts of a compound microscope

How a Light Microscope Works Convex Lenses are curved glass used to make microscopes and glasses etc. Convex Lenses bend light and focus it in one spot.

How the Microscope works Important Terms Field of view - area of the specimen that can be seen through the microscope with a given objective lens Magnification - product of the magnifying powers of the objective and eyepiece lenses. Increases the size Resolution - the closest two objects can be before they're no longer detected as separate objects (usually measured in nanometers) The power to show detail In the 1870s, a man named Ernst Abbe explained why the resolution of a microscope is limited. He said that since the microscope uses visible light and visible light has a set range of wavelengths. The microscope can't produce the image of an object that is smaller than the length of the light wave. Orlando Science Center

RESOLUTION The minimum distance two points can be apart and still be distinguished as two separate points

Depth of Field Focal Plane Depth of field - vertical distance, from above to below the focal plane, that yields an acceptable image Focal Plane Orlando Science Center

Image quality When you look at a specimen using a microscope, the quality of the image you see is assessed by the following: Brightness - How light or dark is the image? Focus - Is the image blurry or well-defined? Resolution - How close can two points in the image be before they are no longer seen as two separate points? Contrast - What is the difference in lighting between adjacent areas of the specimen? Orlando Science Center

Brightness Focus Orlando Science Center

Resolution Contrast Orlando Science Center

Parts of a microscope

Ocular Lens Eyepiece Arm Base Used to support the microscope when carried. Holds the body tube, nose piece and objective lenses Base Magnifies the specimen image Supports the microscope

Objective Lenses Nose Piece Body Tube The body tube holds the objective lenses and the ocular lens at the proper distance The Nose Piece holds the objective lenses and can be turned to change the lens The Objective Lenses increase magnification (usually from 10x to 40x)

Diaphragm Light Source Turn to let more light in or to make dimmer. Projects light upwards through the diaphragm, the specimen and the lenses Some have lights, others have mirrors where you must move the mirror to reflect light Diaphragm Turn to let more light in or to make dimmer. Light Source The Diaphragm controls the amount of light on the slide/specimen

Stage Coarse Adjustment Knob FINE ADJUSTMENT KNOB Moves the stage up and down (quickly) for focusing your image Supports the slide/specimen FINE ADJUSTMENT KNOB This knob moves the stage SLIGHTLY to sharpen the image

Magnification Ocular lenses usually magnifies by 10x To determine your magnification…multiply the ocular lens by the objective lens Ocular 10x Objective 40x  10 x 40 = 400 Ocular lenses usually magnifies by 10x So the object is 400 times “larger” Objective Lens have their magnification written on them.

Caring for a Microscope Clean only with a soft cloth/tissue Make sure it’s on a flat surface Don’t bang it Carry it with 2 HANDS…one on the arm and the other on the base

Carry a Microscope Correctly

Using a Microscope Start on the lowest magnification Don’t use the coarse adjustment knob on high magnification…you’ll break the slide!!! Place slide on stage and lock clips Adjust light source (if it’s a mirror…don’t stand in front of it!) Use fine adjustment to focus

Electron Micrographs

The Scanning Electron Microscope produces a 3-dimensional image of specimen’s surface features spider head of a butterfly

Transmission electron microscope Chloroplast from a tobacco leaf H1N1 virus

THE END