Cell Structure Flashcards
Outline how a student could prepare a temporary mount of tissue for a light microscope
- Obtain thin section of tissue e.g. using ultra tome or by maceration
- Place plant tissue in a drop of water
- Stain tissue on a slide to make structures visible
- Add coverslip using mounted needle at 45* to avoid trapping air bubbles
Describe how light microscopes work
- Lenses focus rays of light and magnify the view of a thin slice of specimen
- Different structures absorb different amounts and wavelengths of light
- Reflected light is transmitted to the observer via the objective lens and eyepiece
Describe how a TEM works
- Pass a high energy beam of electrons through a thin slice of specimen
- More dense structures appear darker since they absorb more electrons
- Focus image onto fluorescent screen or photographic plate using magnetic lenses
Describe how a SEM works
- Focus a beam of electrons onto a specimens surface using electromagnetic lenses
- Reflected electrons hit a collecting device and are amplified to produce an image on a photographic plate
Describe how a laser scanning confocal microscope works
- Focus a laser beam onto a small area on a samples surface
- Fluorophores in the sample emit photons
- Photomultiplier tube amplifies the signal onto a detector. An image is produced pixel by pixel in the correct order
How should the field of view in microscopy be recorded
Draw a diagram with a sharp pencil. Do not use sketchy lines or shading
Include a scale bar
Annotate visible structures
State an equation to calculate the actual size of a structure from microscopy
Actual size = Image size / magnification
Define magnification and resolution
Magnification: Factor by which the image is larger than the actual specimen
Resolution: Smallest separation distance at which two separate structures can be distinguished from one another
Why do samples need to be stained for light microscopes
Coloured dye binds to the structures
Facilitates absorption of wavelengths of light to produce image. Differential staining : contrast between heavily and lightly stained areas distinguishes structures
State the magnification and resolution of a compound light microscope
Magnification - x2000
Resolution - 200 nm
State the magnification and resolution of a TEM
Magnification - x500000
Resolution - 0.5nm
State the magnification and resolution of a SEM
Magnification - x500,000
Resolution - 3-10nm
Explain how to use an eyepiece graticule and stage micrometer to measure the size of a structure
- Place micrometer on stage to calibrate eyepiece graticule
- Line up scales on graticules and micrometer. Count how many graticule divisions are in 100um on the micrometer
- Length of 1 eyepiece division = 100um / number of divisions
- Use calibrated values to calculate actual length of structures
Describe the structure of the nucleus
- Surrounded by a nuclear envelope, a semipermeable double membrane
- Nuclear pores allow substances to enter / exit
- Dense nucleolus made of RNA and proteins assemble ribosomes
Describe the function of the nucleus
- Contains DNA coiled around chromatin into chromosomes
- Controls cellular processes: gene expression determines specialisation & site of mRNA transcription, mitosis, semi conservative