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