microscopy learn how to use a light microscope identify the parts and its functions define...
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Microscopy Learn how to use a light microscope Identify the parts and its functions Define resolution and magnification Identify different kind of cell and its components
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The Microscope
Correct use and care:
transport and lenses
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The Microscope
Each type of microscope has a specific function, which makes them suitable for different types of tasks.
Fig. 1: dissecting, fluorescence, phase-contrast and electron microscopes.
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The Microscope
Ocular
Binocular tube Arm
Revolving nosepiece Objectives
Condenser
Lamp
Base
Coarse focus adjust.Fine focus adjust.
Mechanical stage adjustment (x,y)
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Brightfield
Light is sent up from under the stage Light goes through condenser and is focused
on an object Light goes through object, into objectives and
object is magnified Light goes through oculars to eye and is
magnified again
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The Microscope
Magnification: number of times an object is magnified by the lenses
TM = Ocular x Objective
Resolution: the ability of a microscope to show to separate dots as discrete units
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The Microscope
Lenses:
“Low power" objectives: are very useful to get overview images of larger sections.
“High power" objectives: are mostly for details and there are more delicate (dry and wet lenses).
As the magnification increases in the lenses, the working distance decreases.
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The Microscope
Lenses:
The Immersion lens (100X) is used to increase resolution.
Generally used with oil since its refractive index is very close to that of glass, avoiding lost of light.
The distance between the front lens and the cover glass, also called working distance (WD), can be as small as 0.1 mm.
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The Microscope
Estimating size: we need to determine the diameter of field of view for the objectives and estimate the number of times that the object could fit on the area.
Lo = Df or Lp / # of objects
Lo = length of the objectDf = diameter of field of viewLp = length of the pointer (typically ½ of Df)
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The Microscope
Estimating size:
Lo = Df or Lp / # of objects
Objective Df (mm)
Scanning 3.0
Low power 1.2
High dry 0.3
Oil immersion 0.1
Table 1: standard set of measurements for Df for common objectives of many microscopes.
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Estimating size with microscope
Pointer
Diameter
•Df or Lp/# of objects =Lo
Ex: if Df =0.1mm and there are 2 objects…?
.01mm/2 = 0.05mm/object
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ExampleImage 1: plant cells of Elodea sp. in aquarium water
N
CM
Cl
CV
CW
Plant cells Elodea sp. 400X Elodea sp. cell 0.075mm
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Cell Theory Cells are the basic units of life All living organisms are composed by cells Prokaryotic vs. Eukaryotic cell Animal vs. Vegetal cell
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Biodiversity
Living organisms are organized into
3 domains
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The Cell
Cells are the smallest units that exhibits all the characteristics of living organisms
Fig.2: squematic diagram of prokaryotic and eukaryotic cell
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Biodiversity
Today, we will see examples of 2 of them
2 domains
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The Cell
Prokaryotic cell: Bacteria
Identify rod-shaped and coccus-shaped bacteria
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