Cell structure and Microscopes Flashcards
define resolution
resolution is the minimum distance apart that two objects can be in order for them to appear as separate items
what is the equation for magnification
image size/object size
define magnification
the increase in the size of an object when viewed through a microscope or other optical device
How do laser scanning confocal microscopes work?
they use a laser to only visualise a very narrow region of the sample
how do light microscopes work?
light (source of image) passes through the specimen and focused on an eyepiece which magnifies the image
How do electron microscopes work?
the electron (source of image) beam passes though the specimen and are focused using electromagnetic lenses. the electron beam has a very short wavelength and so has a high resolving power
what is the ability to reproduce detail known as?
the resolving power or resolution
define the details of a light microscope
- focused through a lens
- x1500 magnification
- 0.2 micrometers resolution
- live specimen can be used
- can use both section and external view of specimen
- cheap
- image viewed directly
- staining is required
define details of a transmission electron microscope (TEM)
- focused through EM condenser
- 50 x 10^6 magnification
- 0.1 nm resolution
- live specimen not used
- section of specimen used
- expensive (1.5 million)
- computer screen to see image
- staining is required
define details of a scanning electron microscope (SEM)
- focused through EM condenser
- 1.5 x 10^6 magnification
- 0.02 micrometers resolution
- live specimen not used
- external view of specimen use
- cost in between
- computer screen to see image
- staining is required
what is a stain?
used to increase contrast so that individual subcellular structures can be identified more easily e.g. methylene blue
what is an artefact?
things that you can see down the microscope that aren’t part of the cell or specimen that you’re looking at e.g. dust, fingerprints (created from a stain or the incorrect preparation of a slide)
wavelength - resolution
the shorter the wavelength, the higher the resolution
Limitations of TEM
- must be in a vacuum so no living species
- complex staining process
- not in colour
- section needs to be very thin
- image may contain artefacts
why can’t 0.1 nm always be used as the resolving power for the TEM?
- difficulties preparing specimen limits resolution
- higher energy electron beam required and may destroy specimen
what do we use to mesaure the size of objects when using a light microscope?
Eyepiece Graticule
What is an eyepiece graticule?
glass disc in the eyepiece with scale etched on it (usually 10 mm long with subdivision)
Why can’t a graticule be used to directly measure objects under the microscope?
With different objects, the objective lens will magnify to a different degree so it needs to be calibrated for a particular objective lens
How do we calibrate an eyepiece graticule?
Using a stage micrometer
What is a stage micrometer?
It has a scale etched onto it and it is placed on the stage of the microscope (usually 2 mm long and smallest subdivisions are 0.01 mm)
How do you calculate the scale for different objective lenses (graticule and micrometer)?
Dividing the differences in magnification
What are all the sub cellular structures in a eukaryotic cell?
Nucleus
Mitochondria
Ribosomes
Golgi apparatus
Endoplasmic reticulum
lysosomes
Centrioles
What extra sub cellular structures do plant cells have?
Permanent Vacuole
Chloroplast
Cell wall
What is the shape and size of the nucleus?
Spherical in shape and between 10 and 20 micrometers in diameter
What is the nuclear envelope?
- It’s a double membrane that surrounds the nucleus
- Its outer membrane is continuous with the endoplasmic reticulum and it often has ribosomes on the surface
- Controls the entry and exit of materials in the nucleus
- Contains the reactions taking place within the nucleus
What are nuclear pores?
- Allows the passage of large molecules (messenger RNA)out of the nucleus
- around 3000 pores in each nucleus (40-100 nm diameter)
What is the nucleoplasm?
Granular, jelly-like material that makes up the bulk of the nucleus
What are chromosomes?
They consist of protein-bound, linear DNA (the ends of the DNA are not joined to form a loop)
What is the nucleolus?
- A small spherical region in the nucleoplasm
- Manufactures ribosomal RNA and assembles the ribosomes
- May be more than one nucleolus in a nucleus
What are the functions of the nucleus?
- To act as the control centre through production of mRNA and tRNA (so protein synthesis)
- Retain genetic material of cell in form of DNA and chromosomes
- Manufacture ribosomal RNA and ribosomes
What are chromatins?
- They are the dark patches and they consist of DNA coiled around proteins called histones
- Together they form chromosomes
What is the endoplasmic reticulum?
- An elaborate 3D system of sheet-like membranes, spreading throughout the cytoplasm
- Continuous with the outer nuclear membranes
- The membranes enclose a network of tubules and flattened sacs called cisternae
What are the two types of Endoplasmic Reticulum?
Rough endoplasmic reticulum
Smooth endoplasmic Reticulum
What are the functions of endoplasmic reticulum?
- It has ribosomes present on the outer surface of the membranes
- It provides a large surface area for synthesis of proteins and glycoproteins
- Provides a pathway for transport of materials, especially proteins, throughout the cell
What are the functions of smooth endoplasmic reticulum?
- No ribosomes on surface and often more tubular in appearance
- It synthesises, stores and transports lipids and carbohydrates
What is the Golgi apparatus?
Consists of a stack of membranes that make up flattened sacs, or cisternae, with small rounded hollow structures called vesicles
What happens to the proteins and lipids produced by the ER?
- They pass though the Golgi apparatus in a strict sequence
- They are modified by the Golgi and often have non-protein components added to them (e.g. carbohydrates)
- They are also labelled so they can be sorted and sent to correct destinations
What happens to the proteins and lipids once they are sorted in the Golgi?
- They are transported in Golgi vesicles which are regularly pinched off from the ends of the Golgi cisternae
- The vesicles may move to the cell surface, where they fuse with the membrane and release the contents outside