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CMCB > Lecture One > Flashcards

Lecture One Flashcards

1
Q

Prokaryotic cytoplasm. Eukaryotic cytoplasm.

A

no cytoskeleton; streaming, endocytosis, exocytosis/ cytoskeleton, streaming, endocytosis, exocytosis.

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2
Q

Three major components of cell, in order?

A

water, protein, and RNA.

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3
Q

Smallest component in cell?

A

inorganic ions and miscellaneous small metabolites.

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4
Q

What are the small metabolites?

A

not macromolecules. Amino acids, not proteins, sugars, but not polysaccharides, single nucleosides but not RNA or DNA.

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5
Q

what is a neutrophil?

A

a white blood cell

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6
Q

layers of gut epithelial cells, from apical to basolateral?

A

lumen, epithelium, connective tisssue, circular fibers, longitudinal fibers, connective tissue, epithelium. The epithelium is made up of epithelial cells. The connective tissue is made up of fibroblasts.

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7
Q

What makes up smooth muscle?

A

circular fibers, on apical side, and longitudinal fibers

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8
Q

what are the four major tissue types? What do tissues organize into?

A

connective, epithelial, muscular and nervous tissue. These tissues organize into functioning organs.

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9
Q

what exists between cells in the kidney?

A

the extracellular matrix

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10
Q

what are examples of microscopy?

A

light microscopy, electron microscopy, and atomic force microscopy

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11
Q

What are examples of light microscopy?

A

fluorescence microscopy

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12
Q

Examples of electron microscopy?

A

TEM and SEM

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13
Q

How do antibodies work to help us study cells?

A

provide specificity in conjunction with bionchemical and molecular studies as well as with immunolocalization using microscopy.

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14
Q

what is the role of green fluorescent protein?

A

it helps with cellular localization. It is from the jelly fish. can be artificially engineered. Works genetically just before stop codon.

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15
Q

What are radioisotopes for?

A

they are used for labeling in pulse chase experiments

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16
Q

what is centrifugation?

A

it is used in fractionation of cells. Where we basically divide components of cell by density.

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17
Q

What are bioinformatics?

A

computational biology used for the analysis of large data sets generated in a variety of genome projects.

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18
Q

what does secondary antibody carry to label primary antibody?

A

a label like a fluorescent tag, radioactive atom, gold, enzyme, etc.

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19
Q

How can antibodies help with antigen-antibody complexes form large aggregates?

A

antibodies can selectively precipitate a specific molecule from a complex mixture.

20
Q

polyclonal antibodies

A

collections of antibodies that recognize multiple epitopes of single molecules.

21
Q

How can one produce antibodies that recognize only one epitope? What are those antibodies called?

A

Fuse lymphocytes with immortal cells, and produce single antibody producing cells, called hybridomas. These antibodies are called monoclonal antibodies.

22
Q

Hybridomas are…

A

single epitope recognizing antibodies bred from fusion of lymphocyte with immortal cells

23
Q

What is powerful about GFP?

A

one can attach the GFP sequence to the gene of any protein of interest, and essentially attach a light tag to the protein that can be seen in real time as protein is made and moves via fluorescent microscope.

24
Q

How did scientists determine GFP structure?

A

they crystallized it and determined its amino acid sequence. They discovered where the color was determined in the sequence and can now tweak that location to change color.

25
Q

what is a GFP protein chimera?

A

a sequence encoding GFP is inserted before the stop codon so that the GFP is synthesized in the newly formed, folded protein, which then contains a light tag.

26
Q

what is the limit of resolution of the light microscope? what does that come out to? what can we see?

A

wavelength of visible light, on the order of a few hundreds of nanometers, and allows us to see organelles.

27
Q

what imposes the limit of resolution of the electron microscope? what does that come out to? what can we see?

A

wavelength of the electrons in the electron beam, which is on the order of a few tenths of a nanometer. We can see atoms at 0.2 nm. we can’t see details of molecular structure.

28
Q

what is the minimum resolution we can see with naked eye?

A

0.2 mm or 200 micrometers.

29
Q

With what three different light modulations can we achieve contrast with a light microscope?

A

we can use simple bright field optics, phase-contrast optics, or interference-contrast optics.

30
Q

How are phase-contrast and interference-contrast optics different? how are they the same?

A

the sample will look different. In phase contrast optics, image is dark and highlighted by light. In interference-contrast optics, sample looks more three dimensional. both make use of differences in the way light travels through regions of the cell with different refractive indexes to provide additional contrast.

31
Q

How do simple bright field optics work?

A

white light illumination from beneath the cell

32
Q

What is staining used for? what kind of staining is used in histology?

A

staining can be used to specifically color particular molecular structures. haematoxylin and eosin (H&E) stain is used histologically.

33
Q

What are two ways to use fluorescent labeling?

A

Fluorescent labeling can be used either directly by GFP tagging a particular protein or by linking a fluorescent dye to a specific monoclonal antibody.

34
Q

How does Transmission Electron Microscopy work?

A

a beam of electrons passes through a series of magnetic lenses that focus the bean on the sample and then on the detector.

35
Q

Why does electron microscopy have to be done under vacuum?

A

because the electrons would be scattered by air molecules otherwise.

36
Q

why is electron microscopy not compatible with living cells?

A

electrons cannot pass through thick samples, so samples must be carefully fixed, processed and prepared as very thin sections.

37
Q

How is contrast accomplished in EM?

A

with heavy metals, which block electron transmission more effectively than biological tissue.

38
Q

How can heavy metal atoms be incorporated into EM?

A

two examples. Osmium tetroxide tends to react with lipids and provide high contrast for cell membranes. Gold particles may be attached to antibodies that are specific for a particular cell component.

39
Q

How much better is EM microscopy resolution than light microscopy?

A

TEM can produce images with ~200 times the resolution of light microscopy to allow for the study of cell ultrastructure.

40
Q

How does scanning electron microscopy (SEM) work?

A

the surface of the specimen is imaged by observing the bean of electrons that is reflected by the surface. The sample to be imaged is coated with a thin layer of a heavy metal.

41
Q

What is a nuclear pore?

A

a basket like structure composed of many proteins; its function is to regulate the movement of macromolecuels between the cytosol and nucleus.

42
Q

what is 3D microscopy used for?

A

to study larger three-dimensional structures through the use of serial sections.

43
Q

what are two methods of 3D light microscopy?

A

confocal microscopy and computer-aided image deconvolution

44
Q

what is confocal microscopy?

A

it works in conjunction with fluorescent labels or dyes. a laser light is focused in such a way that it excites fluorescence only at a specific depth in the sample.

45
Q

how does computer-aided image deconvolution work?

A

it removes the bluriness from a series of images focused at different depths

46
Q

What is endoplasmic reticulum contiguous with?

A

the nucleus

47
Q

what makes up cytoskeleton?

A

microtubules, actin filaments and intermediate filaments

CMCB (17 decks)

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