Cell Structure Flashcards
What did Schleiden, 1838 suggest?
All plants are made out of cells
What did Schwann, 1839 suggest?
All animal tissues are compromised of cells
What did Virchow, 1858 suggest?
All cells must arise from preexisting cells
1 cm = ?mm
10mm
1mm = ?um
1000um
1um = ?nm
1000nm
1cm Interm of meters
10^-2m
1mm Interm of meters
10^-3m
1um Interm of meters
10^-6m
1nm Interm of meters
10^-9m
Define magnification
How much bigger a sample appears to be under the microscope than in real life
What is the calculation for total magnification
Total magnification = objective magnification x eyepiece
magnification
Define resolution
The ability to distinguish between two points on the image
How detailed it is
Does increasing the magnification increase the resolution of the image?
No
What is the resolution of a light microscope?
200nm
When can two objects be resolved?
When light can pass between them
When are objects seen as one object?
When the objects are less than 200nm apart
What is a dry mount?
Solid specimens can be viewed whole or as very thin sections
What can be viewed as a dry mount?
Hair, dust, pollen, insects
What is sectioning?
Specimens cut with a sharp blade into very thin sections
What is a wet mount
Specimen suspended in a liquid i.e. water
How is the coverslip placed in a wet mount?
At an angle
What can be viewed as a wet mount?
Aquatic samples etc.
What is a squash slide?
A wet mount prepared and a lens tissue is used to press carefully on the coverslip
What is a squash slide used for?
Soft samples, such as root tips
What are smear slides?
The edge of a slide is used to smear a sample, creating a thin, even coating. The coverslip is then placed on the sample
What are smear slides good for viewing?
Cells in blood
List the ways that a slide can be made permanent?
- Fixing
- Embedding
- Sectioning
- Staining
How/why is fixing used to make a slide permanent?
Chemicals like formaldehyde are used to cross-link proteins, ‘fixing’ the structure of the specimen and preventing chemical breakdown
How/why is embedding used to make a slide permanent?
Some material distorts when you try and cut it into thin sections. Specimens are therefore embedded in wax or resin
How is sectioning used to make a slide permanent?
The tissue is cut using a microtome into very thin sections from 2 to 50um
How/why is staining used to make a slide permanent?
A lot of biological material is not coloured. Some chemicals can bind to the specimen, allowing it to be seen. Some of these ‘stains’ are specific to cell structures e.g. Acetic orcein stains DNA dark red
What stain is used in Gram staining?
crystal violet-iodine complex and safranin counterstain
What colour are Gram-positive bacteria?
Violet
Why are Gram-positive bacteria stained violet?
The presence of a thick layer of peptidoglycan in their cell walls because when they are washed with solvent, the cell prees close, thus becoming less permeable and retain the stain
What colour are Gram-negative bacteria?
Red
Why are Gram-negative bacteria stained red?
The thinner layer of peptidoglycan in their cell walls, and so does not retain the crystal violet whilst in the decolouring process, because the solvent dissolves the lipids, which washes out, or decolourises the stain, safranin, the counterstain is used to stain instead
What is the process of Gram staining?
- Staining with crystal violet dye
- Adding Gram’s iodine solution
- Decolourising the sample
- Counterstaining
What does the first stage of Gram staining involve?
The cells are initially stained with crystal violet dye
What does the second stage of Gram staining involve?
Then a Gram’s iodine solution (iodine and potassium iodide) is added to form a complex between the crystal violet and iodine. This complex is a bigger molecule than the previous crystal-violet stain and iodine and is insoluble in water
What does the third stage of Gram staining involve?
Next, a decolouriser such as ethanol or acetone is added to the sample, which dehydrates the peptidoglycan layer. In a Gram-positive bacteria, the crystal violet-iodine complex cannot penetrate the dehydrated peptidoglycan layer and so is trapped in the cell. However, the external membrane of Gram-negative bacteria is degraded, so the thinner peptidoglycan layer is unable to retain the crystal violet-iodine complex, and the colour is lost.
What does the fourth and final stage of Gram staining involve?
Finally, a counterstain, like safranin, because it is barely water-soluble; is added to the sample, staining it red. The safranin does not affect the violet colouration in Gram-positive cells because it is lighter than crystal violet. Conversely, the decolourised cells are stained red.
What is another name for a laser scanning confocal microscope?
Confocal microscope
What does the laser scanning confocal microscope allow?
Clear observation of whole, living specimens and can focus on structures at different depths within a specimen
What resolution and contrast does the laser scanning confocal microscope images have?
High resolution and contrast
What is the maximum resolution of a laser scanning confocal microscope?
0.8um
How are the images formed in a confocal microscope?
Focusing a single spot of laser light and moving it across the specimen. This causes fluorescence by the components labelled with a dye
What light is detected in a confocal microscope?
Light emitted very close to the focal plane