BIOL 0361 > quiz 1 > Flashcards

quiz 1 Flashcards

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1
Q

what are sample volumes and reagents typically measured in in molecular biology?

A

microliters (1/1000 of milliliter)

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2
Q

what are small volumes measured with?

A

micropipette

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3
Q

draw and label a micropipette

A

-plastic tip
-discharge end
-digital display window
-plunger

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4
Q

what are the steps for using a micropipette (5)?

A
  1. set the desired volume
  2. place tip on the discharge end of the micropipette
  3. depress the plunger until you feel the soft stop and insert tip into the solution just barely below the surface of the liquid
  4. slowly release the plunger; this will draw the liquid into the tip (no bubbles if done correctly)
  5. release the solution into the appropriate container by depressing the plunger to the soft stop, wait one second, and then continue pressing the plunger to discharge the entire volume of the solution
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5
Q

what is a soft stop on a micropipette?

A

the first position the plunger stops at

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6
Q

what is a hard stop used for in a micropipette?

A

for complete discharge of solutions from the tip

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7
Q

what are the 4 rules for micropipetting?

A
  1. never use the micropipette without a tip. This could cause damage to the instrument
  2. gently remove the disposable tips with the ejector button. If necessary, you can remove the tip with your gloved fingers
  3. never lay a loaded pipette or tilt it sideways because this could allow liquid to run back into the instrument
  4. never try to measure a volume that is outside the range of the micropipette
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8
Q

what are the three sizes of micropipette? what color is the tip for each?

A

-1-10 μl (clear)
-10-200 μl (yellow)
-200- 1000 μl (blue)

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9
Q

in the micropipetting lab, what was the x axis on the graph labeled? the y axis? which is the dependent variable and which is the independent variable?

A

-chemical concentration (μM)
-Average % cell viability
-independent: chemical concentration; dependent: average % cell viability

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10
Q

Describe the structure and parts of a compound microscope

A

-illuminator built into the base that light passes through into the substage condeser and adjustable lens system
adjustable lens system focuses light on the object
-iris diaphragm is a lower section of the substage condenser that controls the diameter of the light beam entering the condenser
-two lenses involved in image formation: objectives and oculars
-objective magnify an object from four to one hundred times
-ocular further increases the magnification of the object ten times

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11
Q

what is the resolving power determined by?

A

the limit of resolution

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12
Q
A
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12
Q

what is the limit of resolution?

A

the smallest distance between two points where these points can still be discerned as separate entities

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13
Q

what is the limit of resolution of the naked eye? what is the limit of resolution of a high quality microscope?

A

-0.1 mm
-0.2 μm

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14
Q

what is th formula for the Abbe equation? what does each variable represent?

A
  • limit of reolution=(0.61*λ)/NA
    -λ= wavelength (wavelength of visible light is usually about 600 nm)
    -NA= numerical aperture which is a measure of angle of the cone of light entering the objetive lens.
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15
Q

what happens to the limit of resolution as wavelength of light decreases?

A

the limit of resolution will increase

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16
Q

what happens to resolving power when there is a large numerical aperture?

A

the resolving power is greater

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17
Q

what happens to resolving power when the condenser iris is closed more?

A

the resolving power will decrease despite the contrast and depth of the field being better

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18
Q

what should brightness be controlled with?

A

the on/off switch

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19
Q

what will lowering the substage condenser do to the resolving pwoer?

A

decrease it

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20
Q

what will maintaining a continuous refractive index do to resolving power?

A

greatly increase resolving power

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21
Q

what slide preparation allows us to have the highest resolution power? why?

A

immersion oil because it has the same refractive index of glass, which is different than that of air
-high magnification oil immersion lenses have a small working distance and the immersion oil will connect the objective with the slide without air

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22
Q

what is working distance?

A

the space between the objective and the object being viewed

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23
Q

what are the two major components of blood?

A

plasma and suspended cells

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24
what are the components of plasma (5)?
-H2O -inorganic salts -organic matter -gases -proteins and antibodies
25
what are the components of suspended cells in blood (3)?
-leukocytes (white blood cells) -erythrocytes (red blood cells) -thrombocytes (platelets)
26
how many leukocytes are normally circulating through the blood stream?
3500-12,500
27
what are the two categories of leukocytes? what are the distinctive characteristics of each?
-granulocytes (have conspicuous cytoplasmic granules) -agranulocytes (lack conspicuous cytoplasmic granules)
28
what are the granulocytes (3)?
-neutrophils -eosinophils -basophils
29
what are the agranulocytes (2)?
-lymphocytes -monocytes
30
what is the most frequent cell type?
erythrocytes
31
what are erythrocytes responsible for?
the transport of oxygen to tissues and the transport of carbon dioxide to the lungs
32
where do erythrocytes originate and mature? where are they produced in a diseased state?
-the bone marrow - liver and spleen
33
where can immature red blood cell be found
peripheral blood
34
how are RBCs specialized?
they contain hemoglobin which is used to transport oxygen
35
what is the smallest blood cell? what is its size?
-mature red blood cell -7 μm
36
how do erythrocytes appear in stained preparations?
pink with a lighter central region because of the concavity
37
how long do red blood cells circulate in th eblood stream? where are they terminated?
-120 days -the liver and spleen
38
what does anemia do to red blood cells? give an example.
-distort the size and shape -sickle cell anemia which makes RBCs into a sickle-shape
39
what is the most frequent type of white blood cell? how much of the leukocyte population does it compromise?
-neutrophils -55-75%
40
what are neutrophils generated from?
hematopoietic stem cells and a myeloid precuror cell
41
how are neutrophils recognized when stained with a wright staining prcedure?
-their dark blue, multilobed nucleus -pale pink cytoplasm contiaing small, lightly stained granules and some larger, reddish purple granules
42
why are neutrophils called segmented neutrophils ("segs")
because of their 3-5 lobed nucleus connected by a thin strand of chromatin
43
what are immature neutrophils called? why?
-bands -they do not yet have a segmented nucleus
44
what is another name for neutrophils? why?
-polymorphonucleocytes -because of their varied nuclear morphology
45
what is the shape and size of segs?
round, 9-12 μm in diameter
46
what is the lifespan of neutrophils in the bloodstream?
a few days
47
what is the body's first defense against invading microorganisms?
neutrophils
48
why are eosinophils named in the way they are?
they have a tendency for their cytoplasmic granules to stain with acidic dyes such as eosin
49
what is the size of eosinophils?
10-14 μm in diameter
50
describe a stained eosinophil under the microscope
-dark blue nucleus that might have two lobes (figure eight shaped) -cytoplasm is distinctive; filled with many large, reddish-orange granules
51
what percentage of the leukocyte population is eosinophils?
1-5%
52
how are eosinophils generated?
from hematopoietic cells and a myeloid precursor cell
53
when are increased eosinophil counts seen (2)?
-allergic reactions -in the presence of some parasites
54
why re basophils named the way they are?
due to their tendency to stain with basic dyes
55
what is the most rare leukocyte type? how much of the leukocyte population do they make up/
-basophils -0.5%
56
what is the size of basophils
10 μm in diameter
57
what are basophils generated from?
hematopoietic stem cells and a myeloid precursor cell
58
what do stained basophils look like under the microscope?
-nucleus stains light blue and may be round, banded, or lobed -cytoplasm contains numerous large, deep purple granules
59
what do basophils do?
release histamine during an immune response
60
what is the second most abundant type of white blood cell?
lymphocytes
61
what percentage of the white blood cell population do lymphocytes make up?
20-40%
62
what are lymphocytes generated from?
hematopoietic stem cell and a lymphoid precursor cell
63
what does a stained lymphocyte look like under the microscope?
-nucleus is round and large and stains deep purple -the cytoplasm stains a pale blue; there is only a small layer of cytoplasm surrounding the nucleus in small lymphocytes
64
when are lymphocytes categorized as atypical?
when they exhibit drastic changes in appearance during infectious mononucleosis
65
what are the two types of lymphocytes? what doe each do?
-B-lymphocytes which produce circulating antibodies in an immune response -T-lymphocytes which interact with tumor cells and in viral responses
66
what are monocytes? how large are they?
-large cells -12-15 μm in diameter
67
how much of the leukocyte population do monocytes make up?
3-8%
68
how are monocytes capable of movement?
extensions of the pseudopodia
69
what does a stained monocyte look like under a mincroscope?
-nucleus is round or kidney-shaped, stains lihgtly and does not have many clumps of chromatin -cytoplasm is a dull gray-blue and contains small lilac-stained granules
70
how long do monocytes remain in the bloodstream
1-2 days
71
what is the main function of monocytes?
-as macrophages, phagocytize a variety of foreign substances
72
what are thrombocytes?
-platelets -small, irregualrly shaped cell fragments that have broken away from larger cells in the bone marrow
73
what is the size of platelets?
1-4 μm
74
what do thrombocytes look like under a microscope?
stain purple
75
what is the function of thrombocytes?
play a major role in the blood clotting process
76
what type of stains are Wright stains and Leishman stains? what are they?
-Romanowsky stains -polychromatic stains that re a mixture of eosin and methylene blue
77
what is the color of dye produced based on?
the ionic charge of the stain and the various components of the blood cell
78
what is the charge of eosin ions? what cellular components does it stain and what color?
-negatively charged -basic cell components; orange to pink color
79
what is the charge of methylene blue ions? what cell components do they stain and what color?
-positively charged -acidic components; blue
80
what is the charge on DNA and RNA? what would they be stained by?
-negatively charged -the binding of methylene blue
81
what are the neutral components of cells stained with?
both dyes producing variable colors
82
which staining process is superior for the visualization of cell morphology for red and white blood cells?
lieshman staining
83
what is cell counting used for? what can this information be used for?
-to determine the number of cells in a particular volume -diagnostic testing and experimentation
84
what can trypan blue be used for?
to stain the viable cells and determine the cell viability in a culture sample
85
what is a CBC?
-complete blood count -done by counting the number and type sof different blood cells
86
what is a FACS system?
-Fluorescence-activated cell sorting system -automated cell sorting and counting system
87
what is a hemocytometer?
specialized microscope slide with etched gridlines for ease of counting
88
what is the average number of red blood cells for men? for women?
-4.7-6.1 million cells/mm^3 -4.2-5.4 million cells/mm^3
89
what are all the human blood types (8)?
-A+ -A- -B+ -B- -AB+ -AB- -O+ -O-
90
what determines the four different blood types?
the presence of A and B antigens on the surface of the red blood cell
91
what antigens do those with O blood have?
the absence of both A and B antigens
92
what do the plus and minus in blood type represnt?
the presence of the Rh factor
93
what antibodies would someone with A blood produce?
-antibodies against B antigens
94
which blood type is the universal recipient? why?
-AB+ blood -they don't have antibodies against A, B or Rh because they do not have clotting due to any donated blood
95
which blood type is the universal donor? why?
-O- -they do not have any A, B, or Rh antigens and their blood will not cause a reaction for any patient
96
what are eldoncards? how do they work?
-cards used to determine blood type -the card contains antibodies against A antigens, B antigens, and Rh factor proteins. The blood samples are added and you observe for clotting. The blood sample that contains the antigen will clot with the matching antibody
97
what is the equation used to calculate the number of cells in the solution? explain what each variable means.
RBC count = (Average number of cells x dilution factor x depth factor) / area counted -dilution factor= 200 -depth factor= 10 -area counted= 1/5
97
what will an eldoncard look like for someone with type A+ blood?
-blood will clot in the anti A spot -Blood will clot in the anti-D spot that contains the antibodies for the Rh factor
98
what is flow cytometry?
automated cell counting
99
what is image cytometry?
automated counting of an image
100
what is microfluidic cytometry?
automated counting in solution
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