Background And History 2

Question 1

Contributions of Zacharias Janssen?

Answer 1

> 1859: produced the first operational compound microscope with a magnification of 30x

Question 2

Contributions of Robert Hooke?

Answer 2

> 1665: Robert Hooke described for another cells and introduced the term “cell”

> used an alcohol burner source of illumination and the “bulls eye” lens to focus the light onto the object

Question 3

Contributions of Anthony Leeuwenhoek?

Answer 3

> 1674: Anthony Leeuwenhoek made over 247 simple microscopes capable of magnifications of around 100x. Sent 26 of them to the Royal society. Among his observations were RBCs, Protozoa, striations of skeletal muscle, sperm cells, bacteria

Question 4

Who was credited with the theory that all living organisms are composed of cells?

Answer 4

> Schleiden and Schwann

Question 5

Light waves transmitted through a vacuum do so at a __________ velocity?

Answer 5

> fixed
» velocity is slowed through air, water, or glass
» each medium transmits light at a characteristic velocity

Question 6

How do you calculate the refractive index?

Answer 6

R.I. = (Velocity of light)/(velocity of light inside transmitting medium)

Question 7

What is the refractive index of air?

Answer 7

> 1

Question 8

True or false: when light waves travel from one medium to another, they bend?

Answer 8

> true

Question 9

The degree to which light waves will bend is dependent upon what two things?

Answer 9

> R.I.

> angle at which light strikes the surface

Question 10

True or False: light waves do not follow the same path in either direction?

Answer 10

> False: light waves follow the same path in either direction

Question 11

True or False: light waves striking a perpendicular surface continues on through without bending?

Answer 11

> true

Question 12

What is refractive power?

Answer 12

> a measure of how much a lens bends light waves. It is measured in diopters
»1 diopter = 1 meter divided by the focal length of a lens

Question 13

1 diopter is equal to what?

Answer 13

> 1 meter divided by the focal length of a lens

Question 14

True or False: the thicker the lens, the more that it bends light?

Answer 14

> true

Question 15

What is the focal point?

Answer 15

> the point through which all parallel rays of light will pass after passing through each part of the lens

Question 16

What is the focal length?

Answer 16

> the distance from the center of the lens to the focal point

Question 17

Light waves traveling through which type of lens will converge at the focal point?

Answer 17

> convex lens

Question 18

In relation to the focal point, when is a real image formed?

Answer 18

> a real image is formed when the object is placed OUTSIDE the focal point

Question 19

Summarize a real image

Answer 19

> a real image is formed when the object is placed OUTSIDE of the focal point
» a real image is inverted

> > > a real image can be projected onto a screen

> > > a real image differs in size from the object
»greatest magnification will be obtained from lenses having a very short focal length with the object as close as possible to the focal point

Question 20

When is greatest magnification achieved?

Answer 20

> greatest magnification will be obtained from lenses having a very short focal length with the object as close as possible to the focal point

Question 21

When is a virtual image formed?

Answer 21

> a virtual image is formed when the object placed INSIDE of the focal point

Question 22

When an object is place inside the focal point, what type of image is formed?

Answer 22

> virtual image

Question 23

Summarize a virtual image

Answer 23

> a virtual image is formed when the object is placed inside the focal point
» a virtual image is not inverted

> > > a virtual image cannot be projected onto a screen

> > > a virtual image can be magnified

> > > no points exist at any plane in space at which rays radiating from the object are brought to focus

Question 24

Definition of resolution?

Answer 24

> the ability of a microscope (or any optical instrument) to distinguish two small points as separate points

> to accomplish this, the diameter of diffraction lines around the points must be reduced

Question 25

What equation would you use to calculate resolution?

Answer 25

> resolution = d = 0.61(wavelength of light)/ n sin(alpha)

Question 26

How could you improve resolution?

Answer 26

> use a higher refractive index | > use shorter wavelengths

Question 27

Highest resolution in a light microscope?

Answer 27

> about 0.5 um

Question 28

Highest resolution in the TEM?

Answer 28

> Highest resolution in the TEM (transmission electron microscope) is 0.01 angstroms, but the actual resolution achievable is 5-10 angstroms (= 0.5-1.0 nm)

Question 29

This microscope is also referred to as a bright-field microscope?

Answer 29

> compound optical (light) microscope

Question 30

What are the components of the compound light microscope?

Answer 30

``` > light source > condenser > stage > objective lens > ocular lens ```

Question 31

Pros and cons of the compound optical (light) microscope?

Answer 31

> ability to magnify > ability to solve structural detail > specimen must be thin > relatively little contrast in the unstained specimen

Question 32

Summarize the characteristics of the phase contrast microscope

Answer 32

> converts phase shifts (invisible to the eye) in light passing through a transparent specimen to brightness changes (visible to the eye) in the image > can be used to examine unstinted cells and tissues > use for examination of living cells > different parts of a cell have small differences in the refractive index > light passing through these region becomes deflected and out of phase with the main stream of light waves > out-of-phase wavelengths are matched with other induced out-of-phase wavelengths which cancels their amplitude > this creates light waves that can be seen in contrast to other light waves that have not been retarded

Question 33

Summarize the characteristics of the fluorescence microscope

Answer 33

> detects molecules that emit light of wavelengths in the visible > detects naturally occurring fluorescent (auto fluorescent) molecules such as vitamin A > most widespread application is to detect induced fluorescence >>> detection of antigens of antibodies in immunochemical staining procedure >>> detection of fluorescent tracers injected into animals or cells

Question 34

Summarize the characteristics of the confocal scanning microscope

Answer 34

> patented in 1957 by Marvin Minsky > increases optical resolution and contrast by means of adding a pinhole placed at the confocal plane of the lens to eliminate out-of-focus light > enables the reconstruction of three-dimensional images > combines light microscope components fitted with fluorescence equipment and a scanning system that employs a laser beam > this illuminating laser light is strongly convergent and produces a shallow scanning spot > light emerging from the spot is directed to a photomultiplier tube and analyzed > a mirror system moves the laser beam across the specimen and data from each spot of the specimen is recorded and stored in a computer > the information is the displayed on a monitor to create a visual image

Question 35

What are the advantages to using a confocal scanning microscope?

Answer 35

> very thin optical images of the specimen are created (typically 1 um thick) > out-of-focus images are subtracted from the image by the computer program > computer can make 3D reconstructions of the specimen by stacking individual images

Question 36

Summarize the characteristics of the transmission electron microscope

Answer 36

> TEM uses a beam of electrons rather than light

Question 37

List the components of the transmission electron microscope (TEM).

Answer 37

> cathode * > heated tungsten filament (electron source) > anode * (drives electrons through the column) > series of electromagnets >>> condenser lens, objective lens, projection lens > specimen holder > viewing scree and photographic film *** potential difference between cathode and anode impart and acceleration voltage of between 20,000 and 200,000 volts