Lecture 2 Flashcards
What are the nutrient requirements of cell cultures?
5% CO2 Control of humidity Salts Buffer (typically bicarbonate) Vitamins Amino Acids Glucose Bovine Serum
What are the three sources of primary cell cultures?
Continuous Diploid Cells (taken from embryo), Tumour cells (with oncogenic mutations), Generic cells (often terminally differentiated)
What, in order, are the three steps in Staining for light microscopy
Fixation, Cell immobilization, Staining ot highlight key features
What does phase contrast/ differential interference rely on?
Refractive indexes, the higher the index the darker the image
Why does confocal fluorescence microscopy give a sharper image?
It only takes light from a smaller plane of focus
How do you use fluorescence microscopy to identify individual proteins?
The fluorescent material is linked to an immunoglobin
How do you use fluorescent microscopy to study live organisms?
A chimera protein must be formed, stitching the desired protein together with a naturally fluorescent protein such as GFP
What is detected in a transmission electron microscope?
The intensity of the electron beams that pass through the smaple
What is detected in a scanning electron microscope?
The intensity of the reflected beam
What type of microscopy would you use to in order to gain a 3D type image and gain information about cell surface structure?
Scanning Electron microscopy
What type of microscopy would you use to find out information about the ultra structure of cells?
Transmission electron microscopy?
What is the distinct disadvantage of scanning electron microscopy?
It can not be used in conjunction with other techniques such as immunofluorescent staining
Why is electron microscopy more useful than traditional light microscopy?
Because the smaller wavelength of the electronbeam allows for a greater resolution
