BIOL 303 Study Guide - Spring 2018, Comprehensive Midterm Notes - Protein, Golgi Apparatus, Cell Surface Receptor
BIOL 303
MIDTERM EXAM
STUDY GUIDE
Fall 2018
Chapter 9:
• Recall the scale of cells and subcellular objects (e.g. Fig 9-1 & 9-2, slides 4-5).
• Millimeters to micrometers to nanometers.
• Naked eye (1cm-1.9mm)
• Light Microscope (1.9cm-190nm)
• Electron Microscope (1.9cm-0.1nm)
• Recognize the basic parts of a light microscope (slide 6).
• Eye
• Eyepiece
• Tube lens
• Objective
• Specimen
Condenser
• Iris diaphragm
• Light source
• Recall why the refractive index of media effects microscope image quality (slide 7).
• Diffraction limits brightness and clarity. Resolution is made up of the refractive
index along with wavelength and NA.
• Calculate numerical aperture and effective resolution (slide 8).
• NA = n x sin(a)
• n = refractive index of media.
• a = 0.5 x angle of incidence.
• Resolution = (0.61 x wavelength)/NA
find more resources at oneclass.com
find more resources at oneclass.com
• Briefly describe how fluorophores emit fluorescence (slides 11-12)
• They absorb photons at lower wavelengths and then emit photons of longer
wavelengths and show the color at the wavelength they prefer.
• Briefly describe how antibodies are used to label specific molecules within cells (slide
13).
• Primary antibody: rabbit antibody directed against antigen A which then incubate
cells/tissue in solution containing antibody and the antibody attaches onto antigen
A.
• Secondary antibody: marker-coupled antibodies directed against rabbit antibodies
which then attach onto the primary antibody.
• Recall the light is reflected and transmitted within a fluorescent microscope (slide 14).
• Select wavelengths are reflected down (upright scope) or up (inverted scope)
toward the sample. Then the sample has fluorophores that absorb the wavelength
and emit photons at longer wavelengths. The emitted fluorescence can pass
through the filters that restrict other wavelengths and is directed toward the eye or
the camera.
• Recognize the benefits of each microscopy tool or technique and the scale at which they
operate (chapter 9, slides 6, 14-17,18,22,25-33).
• Light microscope and 200 nanometers apart.
• Fluorescent microscope and 200 nm
• Confocal microscope and 200 nm
• FRET and 200 nm
• STED and 60-70 nm
find more resources at oneclass.com
find more resources at oneclass.com
Document Summary
Iris diaphragm: light source, recall why the refractive index of media effects microscope image quality (slide 7), diffraction limits brightness and clarity. Then the sample has fluorophores that absorb the wavelength and emit photons at longer wavelengths. Immortalized cell lines = primary cells that divide indefinitely: example: through addition of telomerase in somatic cells or a cancerous mutation, 3t3: fibroblast (mouse), hela: epithelial cell (human), 293: kidney(human); transformed with adenovirus, cho: ovary (chinese hamster). Uses a sucrose gradient: both velocity and equilibrium use centrifuge. Low speed (5-600 g) = large objects like entire cells (before lysing) or nuclei sediment out (go to bottom of tube) rest is supernatant (aka sup"). Elution is based on time and/or when the elution buffer is added. Uses an antibody which binds to the proteins within via beads: attach the antibody to the beads and place in the column then add the protein.
