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Biology 360 > Topic 2A - Visualizing cells > Flashcards

Topic 2A - Visualizing cells Flashcards

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1
Q

How small of an object is it possible to resolve with a typical light microscope?

A

200 nm

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2
Q

The naked eye is able to see objects as small as…

A

About 0.3 mm

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3
Q

The very best electron microscopes are able to resolve images as small as…

A

Just over 0.1 nm (a little over 1A)

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4
Q

Who was the first person to build a compound microscope in 1590?

A

Janssen

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5
Q

Who coined the term microscope?

A

Giovanni Faber

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6
Q

The first people to build commercial microscopes with known magnifications were…

A

Zeiss and Abbe

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7
Q

The eyepiece of a typical compound light microscope magnifies this much…

A

10 times

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8
Q

Light waves travelling at slightly different phases which interact with each other can create visual artifacts in a magnified image observed through a microscope. This effect is called…

A

Optical diffraction

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9
Q

Is it possible with current technologies to overcome optical diffraction?

A

No

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10
Q

Two waves IN phase will overlap to create an image that is (bright/dim)

A

Bright

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11
Q

Two waves OUT OF phase will overlap to create an image that is (bright/dim)

A

Dim

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12
Q

What factors influence the limit of resolution?

A

Wavelength of light
Numerical aperture
Medium of the sample

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13
Q

What is the difference between resolution and detection

A 
Resolution = being able to make out a shape
Detection = seeing that something is there

Just because you can see it does not mean you can determine what it is

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14
Q

What are the 4 kinds of light microscopy

A
  1. Bright field
  2. Dark field
  3. Phase contrast
  4. Differential interference
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15
Q

What is the main problem with bright field microscopy?

A

SInce biological tissues are largely uncoloured it is difficult to see any contrast

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16
Q

Describe how dark field microscopy works

A

Add an opaque disc to bright field microscopy, only scattered rays enter the objective

17
Q

Phase contrast microscopy makes what feature of a sample visible?

A

Varying degrees of thickness

18
Q

What is used in a differential interference microscope to obtain a 3D looking image?

A

Polarizer = makes all the light rays go in the same orientation

19
Q

What are 2 methods we can use to detect small objects below the resolution of light microscopes?

A
  1. Electron microscopes

2. Fluorescent microscopes

20
Q

Describe the absorption and emission of fluorescent molecules

A

Absorb light at one wavelength, emit it at a longer wavelength

21
Q

Describe autofluorescence

A

Particles/cells that fluoresce naturally without the addition of fluorescent probes

22
Q

What are the 3 kinds of probes we can fluorescently label?

A
  1. Dyes
  2. Antibodies
  3. Proteins
23
Q

Briefly, how does a fluorescent microscope work?

A

Light comes in, reflected by a dichroic mirror, to point at a sample, sample emits light which is allowed to pass through the dichroic mirror, reaches the eye

24
Q

Describe the outcome of confocal microscopy

A

A stack of images which can be pasted together to create a 3D “scan” of the sample

25
Q

The basis of confocal microscopy is…

A

The 2-pinhole setup to focus light on a very specific plane

26
Q

What are the three methods we can use to detect specific molecules?

A
  1. Dyes detect a class of molecules
  2. Antibodies can detect specific proteins
  3. Genetically engineered proteins detect specific molecules
27
Q

What class of animals produces antibodies?

A

Mammals

28
Q

Why do we use secondary antibodies in biological research?

A

The signal produced by secondary antibodies is 4x stronger than that of the primary antibodies

29
Q

What dye stains genomic DNA?

A

DAPI

30
Q

How can antibodies bind to DNA?

A

Can bind to the histone proteins at the centromere

31
Q

GFP is originally derived from this animal

A

Jellyfish

32
Q

What can be determined from data using genetically engineered fusion proteins?

A

Expression patterns within the cell, can determine what regulatory sequences of a gene have which role

33
Q

At the N-terminus of a protein, an “address” exists, called the…

A

Peptide localization signal

34
Q

What do we detect from using signal peptide marker gene fusions?

A

Intracellular localization of a protein - where it goes in the cell

35
Q

What is the problems with using GFP-like fusion proteins in some settings?

A

Fusion protein can be large which might hinder its function, needs to be translated/transcribed, amplified with PCR… can be tedious

36
Q

Photoactivation experiments typically yield data pertaining to…

A

Protein mobility

Biology 360 (12 decks)

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