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analytical exam 2 > microspoes > Flashcards

microspoes Flashcards

1
Q

when was the first microscope made

A

1590

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2
Q

when was the first time a bacteria was saw

A

1675- with one lens

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3
Q

who wrote a mathematical formula taht correlates to resolving power to the wavelength of light

A

ernst Abbe (Abbe’s principle) in late 80s

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4
Q

when was light made for microscope

A

kohjler oin 1893

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5
Q

when was phase constant illumination made

A

1932- could see through transparent samples with no staining

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6
Q

when was first confocal imaging introduced

A

1957

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7
Q

when was first practical confocal laser scanning microscope made?

A

1978

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8
Q

when was the highest resolution seen

A

1996

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9
Q

what is the focal distance

A

distance from the middle of the lens to the foal point

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10
Q

how to calculate total magnification

A

magnification of eye piece x magnification objective = total magnification

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11
Q

what is an inverted microscope

A

objective under the sample, used for life cell miscopy as cells sink to bottom

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12
Q

what is an upright microscope

A

eye piece on top as well as objective, it can be used for fixed samples. Used for stained cells.

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13
Q

what is a condenser

A

focuses the light onto the specimen, aligns the light rays into a straight path.

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14
Q

out of 63x and 100x which microscope would give the best resolution

A

63

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15
Q

Which lens would you pick if you had multiple colours on your stains

A

plan apochromat

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16
Q

what is refraction

A

the bending of light, occurs when light passes from one medium to another medium with a different refractive index

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17
Q

what is the refractive index

A

a dimensionless number that gives the indication of light bending ability of taht medium

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18
Q

what is the benefit of oil immersion

A

no light is lost so most of information goes in

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19
Q

what is the numerical aperture

A

NA od microscope objective is a measure of its ability to gather light and resolve fine specimen detail at a fixed focal distance

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20
Q

what is fluorescence

A

the properties of some atoms/molecules to absorb light of short wavelength and emit light of longer wavelength

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21
Q

what is stoke shift

A

the distance between the excitation and emission peaks

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22
Q

what is the equation for resolution

A

R= lamba/ (2 X na)

23
Q

how can you show flourcence

A

primary and secondary antibodies with conjugated fluorcence
or can use genetic masses to make them shine

24
Q

what is confocal miscorpsy

A

an advance microscopy method which utilises a pinhole to eliminate out of focus light

25
Q

what are labelling techniques

A

primary and secondary AB with fluoro conjugates and genetic masses

26
Q

what is a multiphore

A

reduce excitation of the laser and bring it to focal planar without exciting other flurorphores so this by using a higher wave length

27
Q

what is confocal microscopy

A

it is an advanced light microscopy method which utilises a pinhole to eliminate out of focus light

28
Q

what numbers are black and white

A

0 is black white and 225 is white

29
Q

what are the different bits of images

A

8 bit- unsigned inter 2^8
16but- unsigned inter 2^16

30
Q

what is the advantage of using LUT

A

used to make the colour in our image match with the wavelength of the light we have detected. Using mulicoloured helps distinguish different shades of the same colour and make pixels more apparent

31
Q

what happens if the object distance is larger than the image distance

A

small and inverted

32
Q

what is the focal point

A

where the light bends

33
Q

what happens if the image distance is further than the object distance

A

image seems bigger

34
Q

how do you calculate total maginifcatoin

A

magbifacation eyepiece x objective = total mag

35
Q

what does inverted microscopy mean

A

object sits under sample- popular in life cell microscopy as sinks to the bottom

36
Q

what is upright magnification

A

eye piece is on top of object

37
Q

what dies the condenser do

A

focuses right on specimen

38
Q

what is plan flour

A

means there are two forms of aberration a chromatic aberration which is wavelength dependant artefacts without corrections would have artefacts

39
Q

which object specification are for one colour

A

plan achromat and achromat

40
Q

what is refraction

A

bending of light, occurs as light passes from one medium into another medium with a different refractive index

41
Q

what are the advantages of immersion liquid

A

has a higher refractive index, no light is lost

42
Q

what is numerical aperture

A

measure of its ability to gather light and resolve fine specimen detail at a fixed focal distance, the higher the better

43
Q

What is white and what is black

A

0=black
225 = white

44
Q

What is confocal light

A

Advanced microscopy which utilises a pinhole to eliminate out of focus light

45
Q

What is an internet value

A

How much each primary colour has been mixed together to create the final colour

46
Q

What is good for qaultive analysis

A

Multichannel/composite images

47
Q

What does successfully extracting information from Microsopy require

A
  • image acquisition
    -image analysis
  • image processing
48
Q

What is deconvulation

A

Corrects the systemic error of blur and reconstructs true image

49
Q

What is Gaussian blur

A

Image is convoluted with a Gaussian function for Smoothing and reducing noise

50
Q

What is thresholding

A

It’s is used for diving image into foreground and back ground

51
Q

What is global threshold

A

Setting cut off pixel value and divide all pixels below and over entire image

52
Q

What is Miniumerror

A

Nonsense

53
Q

What is sedmenTtion used for

A

Find objects and boundaries and quantify their numbers, size, intensity and density

analytical exam 2 (6 decks)

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