Microscopy

Question 1

What is microscopy

Answer 1

Using microscopes to view objects/specimens that are not visible to the naked eye

Question 2

List the parts of the microscope

Answer 2

Detector (photomultiplier, charge-coupled device)
Objective (immersion medium)
Specimen (cover glas)
Light conditioning system
 - Kohler illumination
 - Phase ring
 - Wollaston prism and polarizes
 - Filter cubes (for fluorescence)
Light source (Halogen, XBO)

Question 3

Describe the composition of a light microscopic specimen

Answer 3

Cover glass
Sample surrounded by embedding medium (might contain anti-bleach agent)
Glass slide

Question 4

Describe the control of temperature in a microscopre

Answer 4

Small changes in ambient temperature lead to thermal extension or contraction in the microscope, stage and objective, thereby changing the plane of focus

= incubator box combined w/a precision air heater ensures that the temperature of specimen and microscope remain equilibrated and tightly controlled

Question 5

Describe the box in a microscope

Answer 5

Custom design for the individual microscopy setup. Intricate system of openings and doors for comfortable access to microscope controls and specimen

Question 6

Describe the cube in a microscope

Answer 6

Highest quality fan; controller cube w/external, low-vibration + low-noise design

Question 7

Describe maintenance of CO2 atmosphere

Answer 7

A controller allows to adjust air flow and the %CO2

(there is possibility of guiding the gas stream through a battle of water in order to diminish loss of humidity)

An air tight table top encloses the live cell culture device

Question 8

Problems w/experimental timescales

Answer 8

Artefacts in multichannel/4D imaging

Stability, viability

Question 9

Describe the triangle of frustation

Answer 9

Signal detection (depending on):
Temporal resolution
Sensitivity
Spatial resolution

What is best - depends on application requirements

Question 10

List the markings on objectives

Answer 10

Magnification
Application
Coverslip thickness

Numerical aperture/immersion medium
Working distance

Question 11

Explain what determines objective resolution power

Answer 11

The aperture of the objective determines the resolution. The higher the numerical aperture the better the resolution power of the objective.

RESOLUTION ≠ MAGNIFICATION

Question 12

What is phase contrast of fibroblasts cultured on

Answer 12

Intact collagen

Denatured collagen