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Cell Bio 2382B > Intro to Cell Biology > Flashcards

Intro to Cell Biology Flashcards

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Biology 101 flashcards
1
Q

Cells are supplied with

A
  • Proper nutrients
  • Serum
  • Grown at 37C in CO2 Incubator
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2
Q

Primary Cell Culture

A
  • Divide limited number of times

- Contact Inhibition

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3
Q

Cell Line

A
  • Transformed Cells (HeLa)
  • Immortal
  • Lose Contact Inhibition
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4
Q

Morphology Primary vs Transformed

A

Primary
- elongated + ordered

Transformed
- rounded + disorganized

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5
Q

Symmetric Cell Division

A
  • divides into 2 identical cells from originating
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6
Q

Asymmetric Cell Division

- important for?

A
  • divides into 2 cells uniquely different from originating
  • divides into 2 cells, one is same as originating, one different
  • Stem Cell Renewal
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7
Q

Progenitor Cells

A
  • give rise to differentiated cells
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8
Q

Embryonic Stem Cells

A
  • induced to differentiate into precursors for various cell types
  • pluripotent
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9
Q

Embryonic Stem Cells Procedure

A
  • remove inner cell mass from blastocysts
  • culture ICM on fibroblast feeder cells
  • dissociate surviving ES cells
  • Plate ES cells on new feeder cells
  • allow colonies to form
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10
Q

Adult Stem Cells

A
  • limited differentially
  • located in stem-cell niche
  • adjacent cells provide signals to self-renew or differentiate
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11
Q

Induced Pluripotent Stem Cells

A
  • differentiated cells with Yamanaka Factors
  • Oct4, Sox2, Klf4 = embryonic stem cells
  • c-Myc = cancer cells
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12
Q

Anton van Leeuwenhoek Microscope

A
  • grinding lenses
  • naturally acute eyesight
  • 200x magnification
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13
Q

Bright-Field Microscopy

A
  • light source
  • condenser, objective, ocular lens
  • only high refractive index
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14
Q

Resolution

A
  • ability distinguish between 2 very closely positioned objects as separate objects
  • smaller resolution = better
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15
Q

Resolution Equation

A

D = (0.61 λ)/(Nsinα)

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16
Q

Certain parts of the cell ____ than other parts.
Waves out of phase results in ___ and ____.
Cells with __ refractive properties can ___ passage of a light beam by ___.

A
  • refract more light
  • interference and dim light
  • high slow 1/4λ
17
Q

Phase Contrast Microscopy

A
  • live “unstained” cells
  • annular diaphragm and phase plate
  • 1/2λ reduction – darker contrast
  • phase halo
18
Q

Differential Interference Contrast (DIC) Microscopy

A
  • live “unstained cells”
  • use polarized light
  • defines outline and detail of cell (3D look)
19
Q

Fluorescence Microscopy

A
  • emit visible light when absorb light at specific wavelength
  • excited with fluorochromes
  • instability drops electron releasing energy
20
Q

Fluorescence Microscopy Microscope

A
  • excitation filter – selects wavelength
  • dichroic mirror – reflects excitation and passage of emission light
  • cell specimen labelled with fluorescent dyes that have SPECIFIC excitation/emission wavelengths
21
Q

Fluorochromes

A
  • allow labelling of more than one protein at same time
  • DAPI = blue
  • Mitotracker = red
22
Q

Monoclonal Antibodies

A
  • inject mouse with antigen X = antibodies in spleen
  • fuse spleen cells with myeloma cells
  • transfer to HAT medium
  • unfused cells die (myeloma toxic to HAT, spleen reach Hayflick limit)
  • fused cells are IMMORTAL
23
Q

Immunofluorescence Microscopy

A
  • kill + freeze structures
  • primary antibodies bind to specific protein
  • secondary antibodies (with fluorochrome) bind to primary
24
Q

Fluorescent Imaging in LIVE Cells

A
  • Green Fluorescent Protein
  • excited with blue light = emits green
  • fused to gene of interest on plasmid + transfected
25
Q

Confocal Microscope

A
  • laser excite point of focus

- only excited in focal plane pass through pinhole

26
Q

Deconvolution Microscopy

A
  • math procedure remove fluorescence from out-of-focus parts
  • images take at different focal planes = Z-stack
  • images restored by deconvolution (sharp details)
27
Q

Two-Photon Excitation Microscopy

A
  • two pulsed excitation photons at long wavelength

- image thicker samples

28
Q

Fluorescence Resonance Energy Transfer (FRET)

A
  • 440nm –> 480nm (blue) no protein interaction
  • 440nm –> 535nm (yellow) protein interaction
  • energy from first protein excites the second protein in close proximity
29
Q

Electron Microscopy

A
  • fixed and sectioned samples (NOT LIVE CELLS)
  • specimen stained with heavy metals
  • light –> electron towards anode –> magnetic condenser focuses electrons on specimen
30
Q

Transmission vs Scattered Electron Microscopy

A

Transmission

  • thin sections of cells (intracellular structure)
  • electrons hits specimen, deflect due to heavy metals

Scattered

  • intact cells
  • angle electron beam hits (3D structure)
31
Q

Immunoelectron Microscopy

A
  • primary antibody bind to protein of interest
  • gold particle linked to Protein A bind to primary antibody
  • electron dense spots

Cell Bio 2382B (12 decks)

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