lab test 2 Flashcards

1
Q
  1. What are the components of cell culture environment?
A

o media and media components
o the plastic the cells are attached to
o the gas composition

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2
Q
  1. What are the components of basal cell culture medium? What components you do not expect to see in it?
A

Basal medium contents include water soluble vitamins, trace elements, amino acids. It lacks serum and antibiotics

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3
Q
  1. Do we always add antibiotics to medium?
A

No, only when necessary

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4
Q
  1. What is the purpose of complete medium?
A

To support growth and proliferation of animal cells

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5
Q
  1. What is the purpose of sodium bicarbonate buffer?
A

To help maintain the correct pH when CO2 is present

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6
Q
  1. Is atmosphere inside a biological safety cabinet absolutely sterile?
A

It is very clean but not absolutely sterile

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7
Q
  1. What is the purpose of adding serum into media?
A

o To neutralize toxic nutrients, trypsin and other proteases
o Source of growth and adhesion factors, hormones, lipids and minerals for the culture of cells in basal media
o Regulates cell membrane permeability and serves as a carrier for lipids, enzymes, micronutrients, and trace elements into the cell

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8
Q

How cell surface proteins that promote cell-cell contact will affect cells in culture?

A

Cells stop growing due to contact inhibition

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9
Q
  1. Confluency refers to:
A

The ratio of area occupied by the cells to the total area available

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10
Q
  1. Why cell growth schedule analysis is important?
A

o Allows to establish a cell doubling time
o Allow to see changes in growth patterns over time
o They give record of growth

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11
Q
  1. During the growth of animal cells it is important to keep cells in which phase of growth curve?
A

Log phase

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12
Q
  1. What does PBS stand for?
A

Phosphate buffered saline

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13
Q
  1. The purpose of a disassociation medium (trypsin) is to:
A

To cause adherent cells to detach from one another and the substratum

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14
Q
  1. The purpose of a medium containing FBS after trypsinization is to
A

Reduce further enzymatic activity

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15
Q

Phalloidin selectively binds F-actin. What is F-actin?

A

Filamentous actin

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16
Q

Phalloidin is a poison which affects the depolymerization of actin. A cell treated with phalloidin would likely exhibit difficulties with motility via pseudopodia. True or false?

A

True

17
Q

. Which statement about Rhodamine-Phalloidin staining is NOT TRUE
A. Optimal for fixed and permeabilized samples
B. Very widely cited fluorescent phalloidin conjugate
C. Only used for suspension cell cultures
D. Superior to antibody staining

A

C. Only used for suspension cell cultures

18
Q

. What dye is used in the rhodamine phallodin staining?

A

TRITC

19
Q

Is the excitation wavelength more or less than emission wavelength?

A

Less than the emission wavelength

20
Q

What regions of DNA will Hoechst fluorescent dye most likely bind?

A

A-T regions of DNA

21
Q

What kind of microscope was used to visualize result in the cytoskeleton staining lab experiment?

A

Fluorescent

22
Q

What are the steps of immunochemistry?

A

Cell seeding
Immunostaining
Imaging
Image analysis

23
Q

What is the purpose of using FBS in immunocytochemistry protocol?

A

To block the cells

24
Q

. Name here types of ICC reporters

A

o Enzyme-coupled antibody
o Fluorophore-labeled antibody
o Biospecific small molecule dye

25
Q

You are studying a mouse protein. What would be the least appropriate host to raise primary antibody for this immunochemical experiment?

A

mouse

26
Q

You are studying a rat protein. What is a target specie for a goat anti-rabbit IgG secondary antibody?

A

rabbit

27
Q

Protein Ki-67 is not present in this phase of the cell cycle.

A

G0

28
Q

Compare and contrast DAPI and Hoechst DNA stains.

A

o Both bind to minor groove of double-stranded DNA
o Both bind to A-T region
o Both excited at ultraviolet and emit blue light
o DAPI has greater photostability and can only be used for fixed cells
o Hoechst produces brighter image and can be used for both live and fixed cells

29
Q

Which characteristic of living things is measured in AlamarBlue assay?

A

Metabolism

30
Q

What are the characteristics of the active ingredient of AlamarBlue® (resazurin)?

A

blue and nonfluorescent

31
Q

Is the reagent reduced or oxidized during during AlamarBlue test?

A

Reduced

32
Q

When resazurin is reduced to resorufin, which what color can be observed?

A

bright red fluorescence

33
Q

The amount of fluorescence or absorbance in almarBlue assay is _______________

A

proportional to the number of living cells

corresponds to the cells metabolic activity

34
Q

. Which statement is false about AlamarBlue reaction
A. It is compatible with fluorescence- or absorbance-based instrumentation
B. corresponds to the cells metabolic activity
C. can be used with primary cells or cell lines, with adherent cells and cell suspensions
D. alamar blue is toxic to cells, so this reaction should not be run too long

A

D. alamar blue is toxic to cells, so this reaction should not be run too long

35
Q

. Name at least 3 advantages of using AlmarBlue:

A

Superior Performance
Economical and Safe to Use
Fast & Simple Assay