Cell Structures Flashcards
List the units of length from largest to smallest (with symbol and equivalent in metres)
Describe the one limitation of a light microscope
- long wavelength of light = only distinguish 2 objects if they are 0.2µm apart
How can the limitations of a light microscope be overcome
electron microscope
- uses beam of electrons because of their shorter wavelengths
- therefore you can distinguish objects 0.1nm apart
What is the object
The material under the microscope
What is the image
The appearance of the material when viewed through the microscope
What is the magnification
How many times bigger the image is when compared to the object
What is the equation to work out size of image
Size of image = magnification x Size of real object
What is the resolution, or resolving power
the minimum distance apart that 2 objects can be in order for them to appear as separate items
What does the resolving power depend on
- The wavelength or form of radiation used
What is the resolving power for a typical light microscope
0.2 µm
—> closer than that = will appear as a single item
Does increasing magnification always increase resolution
No
what is cell fractionation
the process where cells are broken up and the different organelles they contain are separated
why is potato homogenised using a cold, isotonic buffer
- Cold: reduce enzyme activity that could break down organelles
- Isotonic: stop water levels being too high/low which would cause organelles to burst/shrink
- buffered: so pH doesn’t fluctuate (changes in pH could alter structure of organelles or affect functionality of enzymes)
what are the 2 stages of cell fractionation
- Homogenation
- Ultracentrifugation
What happens during Homogenation
- cells are broken up by homogeniser (blender)
- releasing organelles from cells
- the resultant fluid = homogenate
- which is then filtered to remove any complete cells and large pieces of debris
where does ultracentrifugation happen
fragments in homogenate are separated in a machine called a centrifuge
what does a centrifuge do
spins tubes of homogenate at high speeds in order to create a centrifugal force
describe the ultracentrifugation process for animal cells
- Tube of filtrate is placed in centrifuge and spun at a slow speed
- heaviest organelles (nuclei) are forced to the bottom of the tube, forming a thin sediment/pellet
- fluid at top of tube (supernatant) is removed, leaving just the sediment of nuclei
the removed supernatant is transferred to another tube and spun faster than before - next heaviest organelle (mitochondria) are forced to the bottom of the tube
- This process continues in a way that at each increase in speed the next heaviest organelle is sedimented and separated out.
what is the speed of centrifugation/ revolutions min-1 for:
- nuclei to be separated
- mitochondria to be separated
- lysosomes to be separated
- nuclei: 1 000
- Mitochondria: 3 500
- Lysosomes: 16 500
why has cell fractionation and ultracentrifugation been useful
allowed detailed study if structure and function or organelles by showing what isolated components do
why are light microscopes considered to have a poor resolution
light has a relatively long wavelength
what are the 2 main advantages of electron microscopes
- electron beam has a shorter wavelength and the microscope can therefore resolve objects well = it has a high resolving power
- electrons are - charge = the charged beam can be focused using electromagnets
what is one of the main disadvantages of an electron microscope
Because electrons are absorbed or deflected by the molecules in the air, a near vacuum has to be created within the chamber of the electron microscope
what are the 2 types of electron microscopes
- Transmission electron microscope (TEM)
- Scanning electron microscope (SEM)
How does a TEM work
- electron gun produces a beam of electrons, which is focused onto the specimen by a condenser electromagnet
- the beam passes through a thin section of the specimen
- parts of the specimen absorbs electrons and therefore appear dark, others allow it through so appear bright
- this produces an image on a screen which is photographed to produce a photomicrograph
What is the maximum resolving power of a TEM, why this may not be easily achieved
Resolving power = 1nm
- difficulties preparing the specimen limit the resolution that can be achieved
- a higher energy electron beam is required and it may destroy the specimen
What are the 4 main limitations of a TEM
- whole system must be in a vacuum = living specimen cant be observed
- complex ‘staining’ process is required, even if the image isn’t in colour
- specimen must be thin
- artefacts may appear on finished photomicrograph that aren’t apart of the natural specimen, therefore we cant assume what we see on a photomicrograph really exists in that form
what are artefacts
things that result from the way the specimen is prepared
–> appearing on the finished photomicrograph
Why must specimen be thin in the TEM
to allow the electrons to penetrate
Does a TEM produce a 3D or 2D image
2D
- we can get over this by creating a 3D image with separate images of different sections of the specimen
–> however this is complicated
How does a SEM function
- directs a beam of electrons onto surface of specimen from above
-beam is passed back and forth across the specimen in a regular pattern - electrons are scattered by specimen
- the pattern of scattering = the contours on specimen’s surface
- we can create a 3D image by a computer analysing these scatterings + secondary electrons produced
Hans and Zacharias Johnson
→ They were eyeglass lens grinders
→ used 2 small convex lenses, found they could magnify to 9x (short focal length)
Robert Hooke
70 years later = created his own version from Onsen’s design to study cork slice = discovering cells
Antony van Leeuwenhoek
Antony van Leeuwenhoek designs hand help microscope
→ can magnify 270 x
→ discovers bacteria and human cells
Richard Zsigmondy
Invents ultra microscope
How do you prepare a slide
- Specimen has to be as thin as possible = let light through
- prepare dry/wet mount
—> stains are used with wet mounts
How to you prepare a wet slide
- Use a pipette, place drop of water onto the middle of a clean microscope slide.
- Place specimen into the water drop using tweezers.
-Place a cover slip on top of the specimen.
(Extra care needed because of trapped air bubbles underneath) - Stand the cover slip upright on its edge over the specimen, then using your tweezers, carefully lower it into place.
- Add any necessary stains after the cover slip is in position.
- Place a drop of the stain at one side of the cover slip.
- Place a paper towel against the edge of the opposite side (drawing the stain under the cover slip and across the specimen)
- You may need to add another drop to ensure the specimen is fully stained, or you may wish to repeat this process with a different stain.
How do you prepare a dry mount
- Place your specimen in the middle of a clean microscope slide using tweezers.
- Place a cover slip on top of the specimen.
- The cover slip will hold the specimen in place and prevent it from getting damaged.
How can we help the image form
Special stains, usually dyes for light microscopy, can be used on specimens like this to help the image form.
What are the 2 most common stains in microscopy
- Common stains include eosin and methylene blue.
- Eosin stains the cytoplasm pink
- while methylene blue stains DNA and RNA blue.
Can multiple stains be used together
Yes - produces best imagery
What is the radiation source of a light microscope, TEM, SEM
- light
- electrons
- electrons
What is the wavelength of a light microscope, TEM, SEM
- 400-700nm
- 0.005nm
- 0.005nm
What is the lens of a light microscope, TEM, SEM
- glass
- electromagnetic
- electromagnetic
What is the specimens used in a light microscope, TEM, SEM
- living or non living supported on glass slide
- non-living supported on a small copper grid in a vacuum
- non-living supported on a metal disc within a vacuum
What is the maximum resolution of a light microscope, TEM, SEM
- 200nm
- 1nm
- 10nm
What is the maximum magnification of a light microscope, TEM, SEM
- 1000 x
- 250 000 x
- 100 000 x
What is the different stains can be used on a light microscope, TEM, SEM
- coloured dyes e.g. methylene blue
- heavy metals
- coated with carbon or gold
What type of image does a light microscope, TEM, SEM produce
- 2D can be coloured
- 2D monochrome
- 3D monochrome
How can we measure the size of objects
Using an eyepiece graticule
What is the graticule
- A glass disc that is placed in the eyepiece of a microscope
- a scale is etched onto this disc
- its typically 10mm long, divided into 100 sub-divisions
- you can see this scale when looking down the eyepiece
Can you directly measure the size of objects under a microscope’s objective lens using an eyepiece graticule
- no
- each objective lens will magnify to a different degree
- the graticule must first be calibrated for a particular lens
- once calibrated in this way, it can remain in this position for future use, provided the same objective lens is produced
How to you calibrate the eyepiece graticule
- use a stage micrometer (special microscope slide)
- this slide has a scale etched onto it that is 2mm long, its smallest subdivisions are 0.01nm
How many units on the graticule scale is 10 units on the micrometer scale
- 40
- therefore 1 unit on micrometer scale = 4 units on graticule scale
- as each unit on the micrometer scale = 10 micrometers, each unit on the graticule:10/4= 2.5 micrometers
If an objective lens magnifying x40 gives a calibration of 25 micrometers per graticule unit, what would an objective lens magnifying x400 mean a graticule unit is equivalent to
25 micrometers / 10 = 2.5 micrometers
What are the general principles for biological drawing
- use sharp pencil only
- use clear, continuous lines
- don’t use any form of shading
- accuracy is paramount - draw what you observe not think
- feint sketching is helpful
- using a magnifying glass or illuminating dissections is useful
- make drawing scaled right
- correct mistakes
- include a title and scale
What are the rules for labelling for biological drawings
- sharp pencil
- label all relevant structures and tissues
- use a ruler for label lines and scale bars
- label lines should touch the subject
- arrange labels neatly (don’t cross-over)
- labels are written horizontally
- title
- scale bar
What are the rules for scale and magnification for biological drawings
- give indication of scale and magnification
- actual sizes are impossible to tell from just a drawing
- if scale and magnification aren’t given = clarify if its a low or high power lens used
- PUT ACTUAL MAGNIFICATION ACHIEVED BY COMBINED EYEPIECE AND OBJECTIVE LENS JUST BELOW THE TITLE
What are the differences between high and low power drawings
- Drawings of cells are typically made when visualizing cells at a higher magnification power,
whereas - plan drawings are typically made of tissues viewed under lower magnifications (individual cells are never drawn in a plan diagram)
What are the guidelines for drawing low power drawings
- draw all tissues and completely enclose each tissue by lines
- don’t draw individual cells
- accuracy is important
What are the guidelines for high power drawings
- Draw only a few representative adjacent cells (3 is sufficient to show enough detail)
- Don’t shade in nuclei - just draw the outline. Similarly with nucleoli
Is this a low power drawing or a high power drawing
Low power
Is this drawing a high or low power drawing
High power
What is the Nucleus’ function
- control centre of cell through production of mRNA and tRNA and hence protein synthesis
- retain the genetic material of the cell in the form of DNA and chromosomes
- manufacture ribosomal RNA and ribosomes
Describe the structure of the nucleus
- Largest organelle
- Spherical
- Dark patches=chromatin
- Surrounded by nuclear envelope
- Composed of 2 fluid filled membranes
- Has nuclear pore-allows large molecules through
- Nucleolus inside
Do prokaryotic cells have a nucleus
No
Why is the nucleus the largest organelle
contains structures like the nucleolus and nuclear envelope that require space to carry out their functions.
What is the nuclear envelope’s function
- double membrane that surrounds the nucleus
- controls the entry and exit of materials in and out of the nucleus
- contains the reactions taking place within it
What is the structure of the nuclear envelope
- its outer membrane is continuous with the endoplasmic reticulum of the cell
- often has ribosomes on its surface
Describe the structure of Nuclear pores
- there are typically around 3000 pores in each nucleus
- each 40-100 nm in diameter
What is the function of nuclear pores
Allow the passage of large molecules, such as messenger RNA, out of the nucleus
What is the nucleoplasm
The granular jelly-like material that makes up the bulk of the nucleus
What are chromosomes made out of
Consist of protein-bound, linear DNA
Describe the structure of nucleolus
- Small spherical region within the nucleoplasm
- there may be more than 1 nucleolus in a nucleus
What is the function of the nucleolus
It manufactures ribosomal RNA and assembles the ribosomes
Describe the structure of centrioles
Small protein tubes of microtubules.
Describe the function of centrioles
Form fibres in cell division known as spindles which separate chromosomes
What stage of cell division is where centrioles are involved
During prophase
Explain why centrioles play an important role
They organise microtubules that serve as the cell’s skeletal system
Describe the structure of the mitochondrion
- rod shaped and 1-10 micrometers in length
- Made up of the following structures:
1. Double membrane
2. Cristae
3. Matrix
Describe the double membrane structure of the mitochondrion
- controls the entry and exit of material
- inner membrane is folded = form extensions called cristae
Describe the cristae structure of a mitochondrion
- cristae are extensions of the inner membrane, which in some species extend across the whole width of the mitochondrion
- large surface area: for attachment of enzymes and other proteins involved in respiration
Describe the matrix structure of the mitochondrion
- makes up the remainder of the mitochondrion
- contains protein, lipids, ribosomes and DNA (allows the mitochondria to control production of their own proteins)
- contains enzymes involved in respiration
What is the function of the mitochondrion
- site of aerobic stages of respiration (Krebs cycle and oxidative phosphorylation pathway)
- production of energy carrier molecule: ATP (from respiratory substances such as glucose)
- number/size of mitochondria and cristae are high in cells that have a high level of metabolic activity (therefore require good supply of ATP)
- e.g. Epithelial cells in intestine require lots of ATP for active transport
Describe the structure of chloroplasts
- vary in shape/size, typically disc-shaped
- 2-10 micrometers long and 1 micrometer in diameter
- consist of 3 main features
1. The chloroplast envelope
2. The Grana/thylakoids/chlorophyll
3. The stroma
Describe the chloroplast envelope structure of chlorplasts
- double plasma membrane that surrounds the organelle
- highly selective in what it allows to enter and leave the chloroplast