3.2- Cells Flashcards
(203 cards)
What is the cell theory?
-All living organisms are made up of one or more cells.
-Cells are the basic functional unit (i.e. the basic unit of structure and organisation) in living organisms.
-New cells are produced from pre-existing cells.
-Cell theory is accepted by ALL scientists!
What is the microscopy equation?
magnification= image size/ specimen size
What are the units of measurement for microscopy?
metre x 1000 = mm
mm x 1000=micrometre
micrometre x 1000= nanometre
Resolution definition?
The minimum distance apart at which 2 seperate objects can be distinguished.
Artefact definition?
Something observed that is only there due to preparation techniques- eg air bubbles.
Magnification definition?
How much bigger the image is than the original object.
What is an eyepiece graticule?
Glass disc with 100 subdivisions, we do not know the length.
What do we do before using an eyepiece graticule?
We have to calibrate the graticule as each objective lens will magnify to a different degree so we cannot use the scale directly.
Have to convert graticule units into real units
How do we calibrate an eyepiece graticule?
You would use a stage micrometer- another slide which is 1mm and has 100 subdivisions- each division is 0.01mm
So how do we measure the size of a specimen?
Line up 1 of the divisions on the eyepiece graticule with a fixed point on the stage micrometer.
Count number of divisions on the eyepiece graticule that correspond with a set measurement on the stage micrometer.
Divide one micrometre measurement (eg 0.1mm) by the n of eyepiece divisions in the same length.
How many µm would each graticule division have if there are 35 divisions in 200µm of the stage micrometer?
length of micrometer in µm/ number of divisions
200 µm / 35 graticule divisions = each division is 5.7 µm
What is cell fractionation?
A method used to isolate different organelles so they can be studied.
How are the cells prepared?
In a cold, isotonic, buffered solution.
Cold- to reduce enzyme activity as when cell breaks open enzymes are released- could damage organelles.
Isotonic- Solution has same water potential as organelles to prevent osmosis as this could cause organelles to shrivel/ burst.
Buffered- solution has pH buffer(stays neutral) to prevent denaturing of the enzyme/protein if the pH were too high or low.
What is 1st step of cell fractionation?
Homogenisation.
Cell must be broken open (homogenised) using a blender/ homogeniser. Cells are blended in cold, isotonic, buffered solution.
Solution is then filtered through gauze to remove any large cell/tissue debris.
What is 2nd step of cell fractionation?
Ultracentrifugation.
Filtered solution is spun at different speeds in centrifuge, organelles separate according to their densities.
What is process of differential centrifugation?
1) Centrifuge spins at lowest speed and centrifugal forces cause organelles to form pellets. Most dense organelles form 1st pellet and move to bottom.
2) Fluid at top of tube (supernatant) is removed, leaving behind the sediment/pellet of the organelle at bottom.
3) Supernatant transferred to another tube and spun in centrifuge at faster speed than before.
4) Process is repeated, with most dense organelle forming sediment, supernatant being spun at faster speed than before.
What is the order that the organelles separate?
Nuclei, Chloroplasts, Mitochondria, Lysosomes, Endoplasmic Reticulum, Ribosomes.
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How does a light microscope work?
Light is focused using glass lenses, it relies on the light being able to pass through the specimen.
Regions that absorb more light appear darker in image.
Advantages + disadvantages of light microscope.
Advantages- easy to use, cheap, show the true colour of the specimen, can use live specimen.
Disadvantages- low resolution due to longer wavelength of light, low magnification (max 1250X)
2 structures in eukaryotic cell that cannot be identified using an optical microscope?
Lysosomes
Ribosomes
What are the 2 types of electron microscope?
Scanning Electron Microscope and Transmission Electron Microscope.
How does SEM work?
It directs a beam of electrons at a specimen.
It creates an image based on the electrons that are reflected.
It is focused using electromagnets to create a 3D image.
Advantages + disadvantages of SEM?
Advantages- much higher resolution than light microscope (1nm), detailed images of surface structures, high mag (300,000X).
Disadvantages- very expensive, extensive training required, samples must be dead (vacuum, stain), false colour/ B+W image.
How does TEM work?
It directs a beam of electrons at a specimen.
It creates an image based on the electrons that are absorbed.
It is focused using electromagnets to create a 2D image.