Light Microscopy Flashcards
What is the smallest creature we can see with a light microscope?
Bacteria (0.5 micrometres)
What is the function of the condenser lens on light microscope?
Focusing the light from the source onto the specimen, concentrating light in a cone shape.
What is the function of the diaphragm on light microscope?
Controls the amount of light passing through the specimen.
What are the parameters that must be optimised in light microscopy?
- Magnification
- Resolution
- Contrast
What are the 5 types of light microscopy?
- Bright field
- Phase contrast
- Differential interference contrast (DIC)
- Fluorescence
- Confocal
What is Bright Field microscope?
Common and basic.
No contrast, do not diffract.
What type of images do phase Contrast microscopes produce?
High contrast images.
What are the two additional components of the Phase Contrast microscope?
Annular ring
Phase ring
What type of images does DIC produce?
3D like, high contrast and detailed
What is the name of the prism that splits the beam in DIC?
Wollaston prism
How does the light path differ between bright-field vs fluorescence microscopy?
Travel through selective filters
What does the first barrier filter do in fluorescence microscopy?
Lets through blue light with wavelength between 450-490nm
What does beam-splitting mirror do?
Reflects light below 510 but transmits light above 510nm.
What does the second-barrier filter do?
Cuts out unwanted fluorescence signals.
Lets through 520-560nm
What is an application for fluorescent microscopy?
Determining the relative localisation of molecules to one another within cells (multiple probes)
