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Bio 201 > 1st week (Lecture 1-4) > Flashcards

1st week (Lecture 1-4) Flashcards

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1
Q

What are intensity profiles

A

method to turn fluorescent microscopic images into graphs

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2
Q

How to make intensity profiles

A
  1. Targeted parts of the cells fluoresce differently by using compounds that bind to specific structures.
  2. Line is then drawn to bisect cell
  3. Graph intensity of colours as a function of location (along the line)
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3
Q

Types of light microscopy

A

Brightfield, Phase Contrast, Differential interference contrast (DIC), Widefield Fluorescence, Confocal fluorescence

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4
Q

Brightfield Microscopy

A

visible light passes through specimen
BRIGHT background
usually needs stains to make cells visible

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5
Q

Phase Contrast Microscopy

A

visible light passes through specimen
more complex lenses, thus better contrast than brightfield
halo around cells

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6
Q

Differential interference contrast (DIC)

A

better lenses than Phase contrast = better contrast
shadow around cells

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7
Q

Widefield Fluorescence microscopy

A

short wavelength hits specimen and longer wavelength reflects off (high to low energy)
needs fluorescent molecules i.e DAPI, fluorescein

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8
Q

Composite images

A

monochromatic images captured, given colour, and superimposed
either additive or subtractive (think of psych)

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9
Q

Confocal fluorescence microscopy

A

lasers reflects off specimen
shows 2D plane within 3D specimen (like a cross section)
more detail than widefield but more expensive

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10
Q

Tomography

A

multiple confocal fluorescence images assembled to make a 3D model

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11
Q

Preparing slides

A
  1. Getting specimen onto slide
  2. Making things visible
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12
Q

Getting things on slide

A

Either the whole cell if small enough or a thin section of a cell/tissue cut with a microtome

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13
Q

Ultrathin Sectioning

A

Done with Microtomes

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14
Q

Coloured Stains

A

2 properties: affinity for target & absorbing light
i.e hematoxylin (blood & RNA), Eosin (proteins)

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15
Q

Fluorescence Stains

A

2 properties: affinity for target & fluoresces light
i.e. DAPI makes nuclei fluoresce blue

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16
Q

Fluorescent Probes

A

2 different molecules joined covalently
1 with affinity for target and 1 fluoresces light
i.e antibodies with Alexa Fluor 488

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17
Q

Light vs Electron Microscope

A

Light: 1000 X, visible or UV light, glass/quartz lens, detected by eye/camera
Electron: 20 000X (SEM) 100 000X (TEM), beam of electrons, electromagnet lens, detected by camera

18
Q

Types of Electron Microscopy

A

Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM)

19
Q

Transmission Electron Microscopy (TEM)

A

Beam of electrons PASSES THROUGH specimen, 2D image

20
Q

Scanning Electron Microscopy (SEM)

A

Beam of electrons BOUNCES off specimen, 3D image

21
Q

Preparing specimens for SEM

A
  • cover specimen (sputter coating) with heavy metal atoms so electrons can bounce off i.e coating with gold
22
Q

Preparing specimens for TEM

A
  1. Ultrathin sectioning + metal atom staining (very common)
  2. TEM tomography (several Ultrathin sections assembled to make a 3D model)
  3. Ultrathin sectioning + (metal atom staining + gold-labelled antibodies covalently attached)
  4. Negative staining
  5. Shadowing (metal atoms sprayed at an angle)
  6. Freeze-fracturing + shadowing
23
Q

Negative Staining (For TEM)

A
  • done when specimen is VERY small
  • stain area surrounding specimen
  • specimen highlighted against a background of metal atoms
24
Q

Freeze-fracturing + shadowing

A
  • see inside of membranes (top or bottom part)
  • specimens are frozen in “freon”
  • cuts through membranes
  • one of the most common
25
Q

What’s part of the Nucleus

A

Nuclear Envelope, Nucleolus & Chromatin

26
Q

What’s part of the Cytoskeleton

A

Microfilaments (Actin Filaments) , Intermediate filaments, microtubules

27
Q

Cell Theory

A

Organisms made of one or more cells
Cell is the basic unit of organisms
cells arise from other cells

28
Q

Cell Specialization

A

i.e flagella (sperm), microvilli (intestine), chloroplasts

29
Q

Unicellular Eukaryotes

A

Paramecium, Green Algae, Yeast

30
Q

Multicellular Eukaryotes

A

Animals: Collagen (protein) as connecting materials
Plants: Cellulose (polysaccharide) as connecting mat.

31
Q

5 categories of Human Cells

A

Epithelial tissue (i.e covering for body surfaces)
nervous tissue (i.e motor neurons)
muscle tissue (i.e skeletal muscle cells)
connective tissue (i.e osteoblasts)
blood cells (i.e red blood cells)

32
Q

Small intestine

A

absorb nutrients (i.e. amino acids, protein, etc.)
Duodenum, Jejunum, Ileum respectively

33
Q

Large intestine

A

recover water before waste is removed

34
Q

Things that increase Nutrient Absorption in the Small Intestine

A

The organ is long and very coiled up
it has folds in the inner membrane
Villi & Microvilli
(all increase Surface Area)

35
Q

Types of intestinal cells

A

Absorptive enterocytes ~90%
Goblet Cells ~10%
Paneth Cells <1%
Enteroendocrine cells (EECs) <1%

36
Q

What are Microvilli filled with

A

Actin Filaments

37
Q

Absorptive enterocytes

A

make up ~90% of intestinal cells.
import nutrients

38
Q

Goblet Cells (secretory cell)

A

make up ~10% of intestinal cells
export protective mucus into gut via secretory vesicles
performs exocytosis

39
Q

Paneth Cells (secretory cell)

A

make up <1% of intestinal cells
export (exocytosis) anti-bacterial proteins into gut
“P”aneth = “P”rotein

40
Q

Enteroendocrine cells (EECs)

A

make up <1% of intestinal cells
export hormones into blood
“entero” = intestine
“endocrine” = secrete into blood

41
Q

Stem cells

A

either reproduce (not by division) or mature into one of the intestinal cells
located at the base of villi and matures upwards

42
Q

How do stem cells mature

A
  1. Reproduce (Self-Renewal)
  2. AE or one of the other 3 secretory cells (EECs, Goblet, Paneth Cells)
  3. EECs or one of the other 2 (Goblet or Paneth Cells)
  4. Goblet Cells or Paneth Cells

Bio 201 (1 decks)

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