Cell structure - Microscopy Flashcards

1
Q

how does a light microscope work

A

the light microscope focuses the light onto the specimen and then magnifies the image through a series of lenses

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2
Q

describe how to use the microscope

A

clip the slide onto the stage
turn on the light
select the lowest powered objective lens
use the coarse adjustment knob to move the objective lens to just above the slide
look down the eyepiece and adjust the focus by moving the lens away from the slide using the fine adjustment knob until a clear image appears

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3
Q

what are stains

A

coloured chemicals that bind to molecules in a specimen

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4
Q

what are the three uses of stains

A

makes cells visible
increases contrast (as different components of the cell take up different amounts of the stain)
detail inside cells can be seen eg organelles can be identified

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5
Q

2 examples of stains

A

methylene blue: stains living cells blue
iodine solution: stains starch grains blue black

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6
Q

describe the process of differential staining

A

using more than one chemical stain. different stains bind to specific structures. this can be used to show contrast between organelles, tissues and microorganisms

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7
Q

define magnification

A

the number of times larger an image is compared with the actual size of the object

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8
Q

what is the equation for magnification

A

M=I/A

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9
Q

define resolution

A

the ability to distinguish between two separate points that are very close together

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10
Q

why is a high resolution good

A

more detail can be seen with a higher resolution, resolution gives clarity

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11
Q

what is the max resolution of a light microscope

A

200nm

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12
Q

what is the max resolution of an electron microscope

A

0.2nm

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13
Q

how do electron microscopes create magnified images

A

by using electrons instead of light waves

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14
Q

why must samples be prepared carefully in an electron microscope

A

to withstand the vacuum inside the microscope

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15
Q

how do electron microscopes work

A

the air is pumped out the microscope and a beam of electrons is fired through a series of electromagnetic lenses which focus the electrons. the electrons are detected and turned into a black and white image (electron micrograph).

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16
Q

why is false colour sometimes added to electron microscope images

A

to make them easier to understand

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17
Q

how must the sample be prepared in a scanning electron microscope

A

sample must be dried carefully and coated thinly in gold

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18
Q

how is the sample prepared in a transmission electron microscope

A

sample must be sectioned thinly

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19
Q

how do SEM work

A

the beam moves back and forth across the specimen. electrons knocked off the specimen are detected

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20
Q

what is the resolution of SEM

A

2nm

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20
Q

what is the maximum magnification of SEM

20
Q

describe the image produced by a TEM

A

2D, high resolution, internal structure

20
Q

how do TEM work

A

use electromagnets to transmit a beam of electrons through a specimen; denser parts of the specimen absorb more electrons, meaning that denser parts appear darker on the final image

20
Q

describe the image produced by a SEM

A

3D, surface of specimen

21
what is the maximum magnification of TEM
x2,000,000
22
what is the resolution of TEM
0.2nm
23
what are SEM used to study
the surfaces of objects
24
what are TEM used to study
the ultrastructure of cells
25
compare the wavelengths of light and electron microscopes
light:400-700nm electron:0.004nm
26
compare the maximum magnifications of light and electron microscopes
light:x2000 SEM:200,000 TEM:2,000,000
27
compare the resolution of light and electron microscopes
light:200nm SEM:2nm TEM:0.2nm
28
compare how focusing is carried out by light and electron microscopes
light: by glass lenses electron: electromagnetic lenses
29
compare the costs to purchase and operate in light and electron microscopes
light: cheap electron: expensive
30
compare the sizes of light and electron microscopes
light: small electron: large
31
compare how materials are prepared in light and electron microscopes
light: easy electron: hard
32
in which microscope is a vacuum required
electron
33
are the materials distorted by the preparation in light and electron microscopes
light: no/maybe electron: yes
34
are the specimens living or dead in light and electron microscopes
light: living or dead electron: dead
35
is colour observed in light and electron microscopes
light: yes electron: no
36
what is an eyepiece graticule
a printed scale in the microscope eyepiece
37
how many units is an eyepiece graticule divided into
100 EPU
38
what is a stage micrometer
a microscope slide which has a fine and accurate scale engraved on it
39
how long is the micrometer scale usually used
1mm
40
how many units is the micrometer scale usually divided into
100
41
how long is each small division on a stage micrometer
10 micrometres
42
does the calibration of the eyepiece graticule have to be done separately for each lens
yes
43
describe simply how to calibrate an eyepiece graticule
line the scale on the stage micrometer with the scale on the eyepiece graticule and count the number of EPU that correspond to 1mm on the stage micrometer.
44
what is the value of one EPU in micrometres for low power objective lens (x40)
25
45
what is the value of one EPU in micrometres for medium power objective lens (x100)
10
46
what is the value of one EPU in micrometres for high power objective lens (x400)
2.5
47
give ways of improving the microscopy practical
use a wet mount cut finely so you can see individual layer of cells squash slide to prevent dehydration/distortion