microspoes Flashcards
when was the first microscope made
1590
when was the first time a bacteria was saw
1675- with one lens
who wrote a mathematical formula taht correlates to resolving power to the wavelength of light
ernst Abbe (Abbe’s principle) in late 80s
when was light made for microscope
kohjler oin 1893
when was phase constant illumination made
1932- could see through transparent samples with no staining
when was first confocal imaging introduced
1957
when was first practical confocal laser scanning microscope made?
1978
when was the highest resolution seen
1996
what is the focal distance
distance from the middle of the lens to the foal point
how to calculate total magnification
magnification of eye piece x magnification objective = total magnification
what is an inverted microscope
objective under the sample, used for life cell miscopy as cells sink to bottom
what is an upright microscope
eye piece on top as well as objective, it can be used for fixed samples. Used for stained cells.
what is a condenser
focuses the light onto the specimen, aligns the light rays into a straight path.
out of 63x and 100x which microscope would give the best resolution
63
Which lens would you pick if you had multiple colours on your stains
plan apochromat
what is refraction
the bending of light, occurs when light passes from one medium to another medium with a different refractive index
what is the refractive index
a dimensionless number that gives the indication of light bending ability of taht medium
what is the benefit of oil immersion
no light is lost so most of information goes in
what is the numerical aperture
NA od microscope objective is a measure of its ability to gather light and resolve fine specimen detail at a fixed focal distance
what is fluorescence
the properties of some atoms/molecules to absorb light of short wavelength and emit light of longer wavelength
what is stoke shift
the distance between the excitation and emission peaks
what is the equation for resolution
R= lamba/ (2 X na)
how can you show flourcence
primary and secondary antibodies with conjugated fluorcence
or can use genetic masses to make them shine
what is confocal miscorpsy
an advance microscopy method which utilises a pinhole to eliminate out of focus light
what are labelling techniques
primary and secondary AB with fluoro conjugates and genetic masses
what is a multiphore
reduce excitation of the laser and bring it to focal planar without exciting other flurorphores so this by using a higher wave length
what is confocal microscopy
it is an advanced light microscopy method which utilises a pinhole to eliminate out of focus light
what numbers are black and white
0 is black white and 225 is white
what are the different bits of images
8 bit- unsigned inter 2^8
16but- unsigned inter 2^16
what is the advantage of using LUT
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
what happens if the object distance is larger than the image distance
small and inverted
what is the focal point
where the light bends
what happens if the image distance is further than the object distance
image seems bigger
how do you calculate total maginifcatoin
magbifacation eyepiece x objective = total mag
what does inverted microscopy mean
object sits under sample- popular in life cell microscopy as sinks to the bottom
what is upright magnification
eye piece is on top of object
what dies the condenser do
focuses right on specimen
what is plan flour
means there are two forms of aberration a chromatic aberration which is wavelength dependant artefacts without corrections would have artefacts
which object specification are for one colour
plan achromat and achromat
what is refraction
bending of light, occurs as light passes from one medium into another medium with a different refractive index
what are the advantages of immersion liquid
has a higher refractive index, no light is lost
what is numerical aperture
measure of its ability to gather light and resolve fine specimen detail at a fixed focal distance, the higher the better
What is white and what is black
0=black
225 = white
What is confocal light
Advanced microscopy which utilises a pinhole to eliminate out of focus light
What is an internet value
How much each primary colour has been mixed together to create the final colour
What is good for qaultive analysis
Multichannel/composite images
What does successfully extracting information from Microsopy require
- image acquisition
-image analysis - image processing
What is deconvulation
Corrects the systemic error of blur and reconstructs true image
What is Gaussian blur
Image is convoluted with a Gaussian function for Smoothing and reducing noise
What is thresholding
It’s is used for diving image into foreground and back ground
What is global threshold
Setting cut off pixel value and divide all pixels below and over entire image
What is Miniumerror
Nonsense
What is sedmenTtion used for
Find objects and boundaries and quantify their numbers, size, intensity and density
