Cells And Microscopy Flashcards
How to complete a dry mount slide?
Solid specimens viewed while or cut into thin slices, sectioning.
Place on centre of slide and place coverslip over the sample
Eg pollen, hair, plant sections
How to wet mount a slide?
Specimen suspended in liquid such as water or immersion oil. Coverslip placed from an angle.
Used for viewing aquatic samples and living organisms.
How to prepare squash slide?
Wet mount prepared first, then Lena tissue is used to gently press down the cover slip
Used for soft samples.
Take care not to damage coverslip when pressing down
How to prepare smear slide?
Edge of slide is used to smear sample creating thin, even coating on another slide, cover slip placed over the sample.
Used for viewing blood.
What is differential staining?
Can distinguish between two types of organisms that would otherwise be hard to identify.
Can also differentiate between different organelles of a single organisms in a tissue sample
What is gram stain technique used for?
Used to separate bacteria into two groups - gram positive and gram negative.
What colour do gram positive bacteria appear under the microscope?
Blue or purple
What colour do gram negative bacteria appear when stained with safranin?
Red
What are the stages of production within slides?
Fixing with chemicals to preserve
Sectioning to form thin slice
Staining for visibility
Mounting on a slide.
What are some rules for a good scientific drawing?
Title
State magnification
Use sharp pencil
Use at least 50% of page
Do not shade
Smooth continuous lines
Clearly defined structures
Label lines should not cross or have arrow heads
What is magnification?
How many times larger the image is than the actual size of the object you’re viewing.
What is resolution?
Ability to see individual objects / structures as separate entities.
What limits resolution?
Diffraction
How can resolution be increased?
By using beams of electrons which have a wavelength thousands of times shorter than light.
Shorter wavelength = beams can be much closer before they overlap
Magnification equation?
Magnification = image / actual
How does electron microscopy work?
Beam of electrons with wavelength of less than 1nm is used to illuminate the specimen.
What can be seen using an electron microscope?
Cell ultrastructure
Up to what magnification can be viewed under electron microscope?
x500 000
What are the 2 types of electron microscope?
Transmission electron microscope
Scanning electron microscope
How does a transmission electron microscope work?
Beak of electrons passed through specimen and focused to produce an image. Has best resolution with resolving power of 0.5nm
How does a scanning electron microscope work?
Beam of electrons passed over the surface of a specimen and reflected electrons are collected.
Resolving power between 3-10nm so not as good
But
3D images of the surface are produced showing us how organisms look.
What is an artefact in microscopy?
A visible structural detail caused by processing the specimen and not a feature of the specimen itself.
What does a laser scanning confocal microscope do?
Moves a single spot of light across a specimen. This causes fluorescence from the components labelled with a dye
How does a confocal microscope work?
Higher light intensity is used to illuminate specimen that has been tested with fluorescent dye. Fluorescence is the absorption and re radiation is light. Light of a longer wavelength and lower energy is emitted and used to produce a magnified image.