Cell Structure Flashcards
it contains the ocular lens
What magnification does the eye piece have? Why?
Has 10x magnification because it contains ocular lens
What can you add/take away from a light microscope that is inserted in the eye piece?
Eye piece graticule can be inserted or taken out (ruler)
What is a turret on a light microscope?
-Contains multiple objective lenses- can be rotated to different lenses
-also known as the revolving nose piece
What is the purpose of the objective lenses on light microscope?
- can be clicked into place by rotating the turret
- contains 4 different lenses= 4 possibilities of magnification to view specimen with
Where is the specimen located on light microscope?
On stage- ensure slide fits in 90* angle
What is the purpose of the stage?
-Specimen is placed onto the stage
- Clips are spring loaded- easily be opened using a lever
What is a condenser in microscopes?
Converges the light rays together to make a beam of focused light
-to pass through aperture in stage
-and then the sample on the slide
What is the iris diaphragm
Underneath the stage above the condenser- size of aperture through which light can pass can be controlled by moving a lever
- by controlling amount of light which passes through slide the specimen can be illuminated but with enough contrast to see details
What is the upper control
Stage clips can be moved backwards+ forwards to position sample
What is the lower control
Stage clips can be moved from side to side to position sample
What is the function of the course focus knob
Bigger/ rough adjustments in height of stage- so sample appears in field of view
What is the function of the fine focus knob
Smaller adjustments- to fine focus the image and view different parts of the same e.g. if 3 dimensions you can focus on each layer
What is the light source in light microscope
Light bulb or mirror
What image does the light microscope form/ produce
2D coloured images
Explain how image is magnified in light microscope
Rays of light pass through sample to form an image—- image is magnified by passing through objective lens times by ocular lens= observer sees magnified image of sample
What is the total magnification in a light microscope
Objective lense times ocular lens equals total magnification
Define microscopic
Items that are too small to be seen with the naked human eye
Define macroscopic
Items that are large enough to be seen with the naked human eye
What is the purpose of microscopes
-Produces a magnified, of the object
-allows biologists to make more accurate descriptions of structures
-give comparisons with other structures
-so we can examine organelles in more details
Advantages of light microscope
-uses light rays to observe object
-can observe living organisms
-does not use harsh chemicals e.g. for staining
-easy to set up and use
-cheap and portable
-colored image
Disadvantages of Light microscope
-low magnification up to 2000 x
-low resolution due to long wavelength of light (200nm)
How to use a light microscope
1) Take a thin slice of specimen- use tweezers to place it in middle of a clean slide
2) Stain the specimen (e.g. eosin to see cytoplasm of cell)
3) Place a cover slip on top of specimen using a mounted needle at a 90* angle
4) Use the arms to clip the slide onto the stage and position so that it sits over the hole
5)Rotate the turret so that lowest objective lens power is in place
6) Use course focus knob to adjust the stage so that it sits underneath the objective lens and look through eye piece to ensure image is roughly in focus
7) Turn fine focus knob to adjust the focus of image until a clear image can be viewed from eye piece lens
8) Rotate the objective lens and select 10 x power and focus image into place using the course and fine adjustment knobs
Define Magnification
refers to how many times larger the image is compared to the object
Define resolution
minimum distance between two objects in which they can still be viewed as separate
-higher resolution= more detail can be distinguished
What determines low resolution in a light microscope
long wavelength of light=low resolution
What determines low resolution in a electron microscope
the wavelength of the beam of electrons
What does TEM stand for
Transmission electron microscope
How is an image created in TEM
-using an electromagnet to focus beam of negatively charged electrons that are fired from the electron gun to pass through sample or specimen
What must be the case for an image to be viewed in a TEM
-specimen must be ultra thin- electrons need to be able to pass through it
- specimen must be dead as it must be viewed in a vacuum
Why are specimens in TEM viewed in a vacuum
Because electrons are absorbed by the air so wont be passed through specimen
How does a TEM function
-extremely thin specimen are stained and placed in a vacuum
-electron gun produces beam of electrons that pass through the specimen
-some parts of specimen absorb electrons- makes them appear darker
-2d image produced showing in detail internal structures of cell
What image does TEM produce
2D and Black and White images
Advantages of TEM
-high magnification (up to 5 million x)
-high resolution
-can see details inside the cell
Disadvantages of TEM
-can only see dead material
-harsh chemicals used in preparation which can cause artefacts
-expensive
-B&W images
-requires skilled, trained individual to complete and prepare specimen
What is the formula involving magnification, image size and actual size
I AM
how to do you go from M to mm
times by 1000
how do you go from mm to M
divide by 1000
how do you go from mm to micrometers
times by 1000
how do you go from micrometers to mm
divide by 1000
how do you go from micrometers to nanometers
times by 1000
how do you go from nanometers to micrometers
divide by 1000
What does SEM stand for
Scanning Electron Microscope
How does a SEM function
-stream of electrons are sent across surface and reflected back to capture image - electrons scatter in different ways depending on the contours of the image
-electrons rebound (bounce off to fluorescent screen- instead of eye piece)
-observed in a vacuum-specimen must be dead
Why does the specimen in SEM not have to be ultra thin
Electrons are not transmitting through they bounce off of image to fluorescent screen
SEM produces what type of image
3D surface image + B&W
Advantages of SEM
-high magnification up to 5 million times
-high resolution
-can see details of the surfaces of structures
Disadvantages of SEM
-can only see dead material
-harsh chemicals used in preparation which can cause artefacts
-expensive
How does a Laser scanning confocal microscope function
-image created using very high light intensity (laser beam of light) to illuminate the specimen stained with a fluorescent dye- these then fluoresce
-image is created as microscope scans the specimen point by point using a focused laser beam to create a 2D or 3D image in different focal planes
Advantages of Laser scanning confocal microscope
-enables scientists to view sections of tiny structures that would be challenging to physically section e.g. embryos- 3D images created
-combines the benefit of high resolution optical imaging with depth selectivity
-can observe cell processes by tracking molecules
-higher magnification + resolution than light microscope
Disadvantages of Laser Scanning Confocal Microscope
-more expensive than light microscope
-more complex than light microscope
Why do we stain things
-to see things much easier and we can differentiate between different parts of the cell
What is dry mount
-sample is cut into a thin slice using a blade- sectioning
-it is covered with a cover slip then is ready to be viewed e.g. hair, pollen, insects
What is a wet mount
-specimen is covered with a drop of liquid e.g. oil or stain or water
-cover slip is placed from an angle using a mounted needle on the sample before viewing it- to avoid bubbles
What is a squash slide
sample is squashed between slide and coverslip- to ensure you have a thin layer of specimen so light can pass through it
- a type of wet mount
-use for soft tissue e.g. root tips squash sample to view chromosomes in mitosis
What is a smear slide
-using another slides edge you smear the sample across a slide- to create a smooth, thin, even coated specimen
- a cover slip is placed on top after smearing
-useful for a thin layer of a liquid e.g. blood sample to view RBC
What is differential staining
-various chemical stains used to stain different parts of a cell in different colors
- used to differentiate between different types of cells that otherwise can not be distinguished
What is an example of a common use of differential staining
Gram staining
What is Gram staining
2 stains used= crystal violet+ safranin
-gram positive bacteria appear blue or purple- stain is retained by thick peptidoglycan cell wall absorbing dye
-gram negative bacteria cannot absorb crystal violet- peptidoglycan cell wall is too thin and do not retain the stain
What is nigrosin and congo red used for
negatively charged- attracted to outside of cell- can’t enter cell as cystol repel them- stained background so unstained cell stands out- can be seen
What is crystal violet and Methylene blue
positively charged so attracted to negatively charged components e.g. organelles
What are the 2 pairs of commonly used stains
1) Nigrosin and Congo red
2) Crystal violet and Methylene blue
What organelles are found in an animal cell
-Nucleolus
-Nucleoplasm
-Nuclear membrane
-Nuclear pores
-Nucleus
- Smooth endoplasmic reticulum
-Rough endoplasmic reticulum
-Secretory Vesicles
-Cytoskeleton
-Cytosol
-Cell surface membrane
-Mitochondria
-Ribosomes
-Golgi Apparatus
-Vesicles
-Centrioles
- Microtubule network
-Lysosomes
-Chromatin
Organelles found in plant cells
-Chloroplast
-Vacuole containing cell sap
-cellulose cell wall
What is the function of nucleus
-contains linear DNA tightly wounded around histone proteins to form thread like structures called chromosomes
What is the function of the Nuclear Envelope
-double membrane that surrounds the nucleus and has nuclear pores
What is the function of the nuclear pores
holes in the nuclear envelope that allow mRNA to leave the nucleus
What is the function of the nucleoplasm
granular, jelly like material contains nucleolus and chromosomes
What is the nucleolus
-largest structure within nucleus
-where rRNA (ribosomal RNA- a single stranded polynucleotide is synthesised at high speed
Define transcription in terms of how a ribosome is formed
process of making RNA copy of a genes DNA sequence- after the rRNA associate with ribosomal proteins they form ribosomal subunits that later assemble to form functioning ribosome
What is the structure of a ribosome
-small+ spherical
-made up of 2 sub-units of proteins and rRNA