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Proc 180 > microscopes lo3-lo4 > Flashcards

microscopes lo3-lo4 Flashcards

1
Q

review kohler in lab

A
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2
Q

review microscopes compound labels

A
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3
Q

a compound microscope is a what type of lens system

A

2 lens system

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4
Q

a microscope magnifies what

A

strucutures

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5
Q

What is the lens and made of

A

optical element
glass or other transparent material

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6
Q

objective produces 4 prts image

A

initial
enlarged
real
inverted image

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7
Q

occur eyepiece produces 3 prts image

A

final
enlarged
virtual

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8
Q

condenser system 2 prts

A

-focuses light rays on the object
-not involved in making the image but affects quality

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9
Q

the illumination system contains

A

a light source

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10
Q

3 mechanical support system, list and purpose

A

-used for focusing and moving stage
-coaxil, fine, course

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11
Q

the principle of the compound microscope, type of image produced

A

-light is directed to the condenser and passes through compound lens system
-image is against a bright background (structure is darker, staining)

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12
Q

the image formed by the compound is due to

A

absorption, diffraction, and refraction of light rays by different parts of the specimen

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13
Q

eyepiece does what from the images of objective

A

re-magnifies the image formed by the objective and produces a virtual image

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14
Q

diopter adjustment

A

allows for individual adjustment of focus for each eye

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15
Q

body

A

hols the objective and eyepieces in proper alignment and at the correct optical distance

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16
Q

revolving nose piece

A

holds the objectives and allows he chnging

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17
Q

objectives produce what image

A

produces initial, magnified, real image

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18
Q

stage coaxial controls

A

moves the slide back forth and side to side

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19
Q

mechanical stage

A

object is places

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20
Q

vernier scale

A

physical position

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21
Q

condenser

A

lense system focuses light into pencil cone and illuminates objects

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22
Q

iris diaphragm

A

reduces spherical aberrations, controls size of lightq

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23
Q

condenser control know

A

up and down condenser, sharpens image

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24
Q

coarse/ fine adj

A

fine: final focus
coarse: initial focus, rapid

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25
Q

light source

A

supplies light

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26
Q

base and arm

A

physical support

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27
Q

slide holder

A

clamps down onto slides

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28
Q

swing out the knob of the condenser front lens

A

allows larger object fields to be illuminated in conjunction with low power objectives

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29
Q

field diaphragm

A

controls the amount of light from light sources, determines the size of the field the light is able to illuminate

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30
Q

3 type of objective lenses and purpose

A

low power= initial focusing
high-dry= larger objects
oil immersion = very small objects

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31
Q

black ring =

A

oil

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32
Q

monocular vs binocular

A

how many eyes can see

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33
Q

condenser larger def

A

Serves to collect the light from the light source, focus the light on the same plane as the specimen and to allow the light the pass through the centre of the object

-condenser should have the same numerical aperture as the objective that is being used

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34
Q

Iris diaphragm

A

built into the condenser and is an adjustable shutter
that controls illumination of the object by altering the size of the cone of light

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35
Q

Filters

A

function to reduce the intensity of light and are usually blue-green

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36
Q

Vernier scale

A

may be present on the mechanical stage and used to
relocate a particular object on a slide

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37
Q

Micrometer scales

A

may also be present to allow for the measurement (size) of
objects under the microscope (ocular micrometer and stage micrometer)

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38
Q

the light source bulb is usually 3 things

A

low voltage
high intensity
tungsten-halogen bulb

39
Q

ocular micrometer , found where and scale

A

-fits into eye piece
-100 equal divisions, uses stage micrometer to determine distance between each line

40
Q

scale micrometer, where is it and scale

A

-on the slide with the scale on it
-scale is 1mm long and divided into 0.01 mm

41
Q

how does the light travel/ path in compound microscopes

A

straight pathway

42
Q

characteristics of light, 3 parts

A

-transmitted in waves
-spreading out in increasing circles
-circles cause crescent and troughs

43
Q

wavelengh is in what units

A

nm

44
Q

wavelength is equal to what

A

from the beginning of a crest/rough to the end of the next crest/trough

45
Q

shortest to longest waves color

A

purple blue green yellow red black

46
Q

what is the amplitude of a wave

A

height of the wave from the zero point to the tip of the crest

47
Q

Greater amplitude =

A

brighter light

48
Q

(not sure if need to know the 4 def)
absorption
reflection
dispersion
refraction

A

– the reduction of light intensity as it is transmitted through an object
– the light that is
reflected back from a surface
– the effect of white light being separated into its component colours
– the bending of light as it passes from one medium to another

49
Q

Refractive Index (R.I.)

A

– amount of bending
that takes place as a ray of light passes
from one substance to another with a
different optical density

50
Q

Resolution
is the ability to
which number is better
controlled by 2 things

A

-Ability to discriminate detai
-Lower number is better! = shortest distance you can discriminate objects
-Resolution is controlled by:
-Wavelength (shorter wavelength = better resolution)
-Numerical aperture of the lens (larger NA = better resolution)

51
Q

Aperture –

A

– opening through which light passe

52
Q

Numerical Aperture (NA)

A

– measure of the fraction of a light wave admitted
by a lens
-The larger the numerical aperture, the more efficient the resolving power of the microscope
-different in each objective

53
Q

Condenser NA should be equal to

A

objective NA

54
Q

We do not want to control light by adjusting the iris diaphragm because this

A

will affect the NA and therefore the resolution of our microscope

55
Q

OIL IMMERSION

A

The Refractive index of glass and oil are the same, therefore no light lost because the light rays no longer must pass through air
-More light entering gives better resolution

56
Q

Magnification, how to calculate total

A

-the degree of enlargement
-Total magnification is a product of the ocular magnification and the objective magnification

57
Q

Visibility

A

– the ability to see an object

58
Q

contrast

A

relative difference in the brightness between the object and its surroundings

59
Q

Working distance

A

clearance between the lowest part of the objective and the object in focus

60
Q

Field of view/Field diameter

A

area of the object that is seen in the microscope

61
Q

Depth of focus

A

– the depth range in which objects can still appear in focus (oil immersion will have basically a zero depth of focus since we can only see focused objects in one plane)

62
Q

Parfocal/Parcentral

A

– the ability of different objectives to focus at the same
position with minimal adjustment

63
Q

The most important components of the microscope!

A

lenses

64
Q

Lenses

A

are glass or similar transparent material with one or two curved surfaces

65
Q

Positive lenses

A

cause light to converge (come together) and magnify
objects

66
Q

negative lenses , cause and do not

A

cause light to diverge (spread apart) and do not
magnify objects

67
Q

Real image is formed by

A

the objective lens system in the focal plane
of the ocular lens

68
Q

Virtual image is formed by the

A

magnification of the real image by
the ocular lens system

69
Q

Focal point is the point where the

A

light rays converge after being
refracted through a convex lens

70
Q

Focal length is the

A

distance from the centre of the lens to the
focal point

71
Q

Aberrations are

A

defects in lenses that prevent us from seeing a
perfect image

72
Q

2 types of abberration

A

Chromatic aberration
Spherical aberration

73
Q

CHROMATIC ABERRATION, corrected, fixed

A

The inability of a lens to converge white light at one point after it has
been refracted by a convex lens creates a colored fringe around the
object know as chromatic aberration

-blurriness and rainbow edge in areas of contrast.

-Corrected by combining positive and negative lenses of different material

Fixed using two types of lenses:
1. Achromatic lens – blue and red colors are fixed
2. Apochromatic lens – red, blue and yellow colors are fixed

74
Q

SPHERICAL ABERRATION, corrected

A

-The inability of a lens to converge all the light rays passing through to a single
point (due to different thicknesses in different areas of the lens) creates a
blurred image or a curved field - “muddy edges”

Can be corrected two possible ways:
1. Block light rays that would normally fall on the marginal areas of the lens
2. Combine a strong positive (convex) lens with a strong negative (concave) lens, the same as chromatic
aberration correction

75
Q

Two important criteria to be met when setting Kohler:

A
  1. Field of view should be evenly illuminated
  2. When iris diaphragm is open and the condenser is in the upper position, the back lens of the objective should be filled with light as completely as possible
76
Q

purpose of kohler

A

-Provides proper illumination for the
microscope and allows for best
possible resolving power and
optimum specimen contrast
-Results in maximum use of the light
source

77
Q

Interpupillary distance, typical value

A

-the distance between the microscope
eyepieces when only one image is seen by both eyes
-Typically would be between 55-75 mm

78
Q

other microscopes lo4

A
79
Q

darkfield microscopy note 3prt

A

Oblique illumination, special condenser, based
on diffraction of light

80
Q

darkfield microscopy specimen view

A

Bright object against dark background (no
intracellular detail)

81
Q

darkfield microscopy uses

A

Very small bacteria and spirochetes;
specimen is unstained

82
Q

phase contrast note

A

Specimens (and their internal structures) can
diffract and refract light, causing changes in
amplitude that we can view as varying
brightness; uses annular diaphragm and phase
plate/ring

83
Q

phase contrast speciment view

A

Positive – dark image on a light background,
intracellular detail

Negative – bright image on a dark
background, intracellular detail

84
Q

phase contrast uses

A

Visualization of internal structures;
specimen is living and unstained

85
Q

Fluorescence note

A

UV light source “hits” specimen with light,
fluorochrome dye fluoresces light where it is
attached; uses excitation filter and barrier filter

86
Q

Fluorescence specimen view

A

Object that is attached to fluorochrome
colour based on fluorochrome and type of
light used against dark background

87
Q

Fluorescence use

A

Detection or identification of
certain tissues; bacteria, viruses,
fungi

88
Q

polarizing note

A

View objects based on their ability to change
direction of polarized light; uses polarizer and
analyzer to create and detect polarized light

89
Q

polarizing specimen view, also called 3 other

A

Bright object against dark background
(objects are also known as anisotropic,
birefringent, doubly refractile)

90
Q

polarizing uses

A

Tissue lipids, myelin sheath,
crystalline lipids in tumours

91
Q

inverted note

A

Illumination system is placed above the
specimen, objective lenses are placed below;
allows for visualization of larger objects

92
Q

inverted specimen view

A

Bright object against dark background

93
Q

inverted uses

A

Cell cultures, fluids, agglutination
tests; specimen is unstained

Proc 180 (4 decks)

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