Practicals Flashcards
TOPIC 1A - (1) What does Catalase catalyse?
It catalyses the breakdown of hydrogen peroxide into water and oxygen
TOPIC 1A - (1) How do you measure the oxygen released in the experiment?
The oxygen displaces water from the measuring cylinder so you measure the empty part of the cylinder
Topic (1) What could you use to hold the cylinder upside down?
A stand and a clamp
TOPIC 1A - (1/2) What are the two ways of measuring the rate of an enzyme controlled reaction?
1 - You can measure how fast the product of the reaction is made (1)
2 - You can measure how fast the substrate is broken down (2)
TOPIC 1A - (2) What does amylase catalyse?
The breakdown of starch to maltose
TOPIC 1A - (general question) MAKE SURE YOU CAN INTERPRET GRAPHS OF ENZYME CONTROLLED REACTIONS. LOOK AT QUESTIONS ON THEM
EXAM MATERIAL
TOPIC 1A - (general question) How can you use a tangent to calculate the rate of a reaction?
- Draw a tangent at the point you want to find out the rate at
- Work out the gradient at that point (change in y/change in x)
Topic 2A -(1) Describe how you can observe mitosis using root tips (Step 1-5)
1- Cut 1cm from tip of growing root (growth occurs here so mitosis also happens here). If you’re using ethano-orcein to stain the cells, the tips will also need to be fixed in ethanoic acid
- Prepare a boiling tube containing 1M HCL and put in water bath at 60 degrees.
- Transfer root tip into boiling tube and incubate for 5 mins
- Use pipette to rinse the root tip well with cold water and then leave the tip to dry on a paper towel
- Place the root tip on a microscope slide and cut 2mm from the very tip of it. Get rid of the rest`
Topic 2A - (1) Describe how you can observe mitosis using root tips (Step 6-9)
(DIAGRAM)
6- Use a mounted needle to break the tip open and spread the cells out thinly
- Add few drops of stain and leave for few mins. The stain will make chromosomes more visible under microscope. Stains include toluidine Blue O, ethano-orcein, feulgen stain (if using feulgen, you’ll need an extra rinse)
- Place cover slip over cells and push down firmly to squash tissue. This will make tissue thinner and allow light to pass through it. DONT smear cover slip sideways or you’ll damage the chromosomes)
- Now look at all stages of mitosis under microscope
TOPIC 2A - Describe how to observe cells using an optical microscope
1- Clip the slide you’ve prepared onto the stage
2- Select the lowest-powered objective lens (one with lowest mag)
3- Use course adjustment knob to bring stage up to just below objective lens
4- Look down eyepiece (which contains ocular lens). Use course adjustment knob to move the stage downwards, away from objective lens, until the image is roughly in focus
5- Adjust focus with fine adjustment knob, until you get a clear image of what’s on the slide.
6- If you need to see the slide with a greater mag, swap to a higher-powered objective lens and refocus
Topic 2A - what is the mitotic index?
The proportion of cells undergoing mitosis
Topic 2A - How do you calculate mitotic index?
Mitotic index = number of cells with visible chromosomes/total number of cells observed
Topic 2A - What does mitotic index tell us?
- It lets you work out how quickly tissue is growing and if there’s anything weird going on
- A plant root tip is constantly growing so you’d expect a high mitotic index
- In other tissue samples, a high mitotic index could mean that tissue repair is taking place or that there is cancerous growth in the tissue
Topic 2A - What should you include in a microscope drawing?
- The mag that the specimen was viewed under
2. Label the drawing
Topic 2A - (2) What could you use to calculate the size of cells
A Graticule and a micrometer
Topic 2A - (2) What is an eyepiece graticule and where is it fitted?
It’s like a transparent ruler with numbers, but no units and it is fitted onto the eyepiece
Topic 2A - (2) What is a stage micrometer, where is it placed and what is it used for?
- It is a microscope slide with an accurate scale (with units) and it is placed on the stage
- It is used to work out the value of the divisions on the eyepiece graticule at a particular magnification
- This means that when you take the stage micrometer away and replace it with the slide containing your tissue sample, you’ll be able to measure the size of the cells
Topic 2A - (2) Give an example of how the graticule and micrometer could be used
1- Line up the eyepiece graticule and the stage micrometer
2- Each division on micrometer is 0.1mm
3- At said mag , 1 division on stage micrometer is the same as 4.5 divisions on graticule (example)
4- So 1 division on graticule is 0.1/4.5 = 0.22mm
5- So if you look at the microscope at that mag and its 4 eyepiece divisions long, you’ll know it measures 4*0.22 = 0.88mm
Topic 2A - (1) What are artefacts?
Artefacts are things that you can see down the microscope that aren’t part of the cell or specimen that you’re observing (including dust, air bubbles, fingerprints etc
- They are usually made during the prep of your slides so you must prepare your root tip cells carefully to avoid them
Topic 2A - Why are artefacts especially common in electron micrographs
- Because specimen need a lot of prep before you can view them under an electron microscope
- The first scientists to use these microscopes could only distinguish between artefacts and organelles by repeatedly preparing specimens in different ways. If an object could be seen with one prep technique but not another, it was more likely to be an artefact than an organelle
Topic 2B - (1) Describe the practical method to investigate how temperature affects beetroot membrane permeability
1) Use a scalpel to carefully cut 5 pieces of beetroot (on a cutting board). Rinse the pieces to remove any pigment released during cutting
2) Add the 5 pieces to 5 diifferent test tubes, each containing 5cm^3 of water (use pipette or measuring cylinder to measure water)
3) Place each etst tube in a water bath at a different temperature e.g. 10, 20, 30, 40, 50 degrees for the same length of time. (measure time with stopwatch)
4) Remove the pieces of beetroot from the tubes, leaving just the coloured liquid
5) Use a colorimeter. The higher the absorbance, the more pigment releaser, so the higher the permeability of the membrane
6) You can connect colorimeter to a computer and use software to collect the data and draw a graph of results