Introduction to Microscopy Flashcards
What is microscopy
Using microscopes to view objects/specimens that are not visible to the naked eye.
Name the parts of a microscope
- Light source
- Light conditioning system
- Specimen
- Objective
- Detector
How is the light microscopic specimen prepared
- use a coverglass of thickness 0.17mm
- Sample is surrounded by embedding medium
- use a glass slide
Explain how life imaging microscopy works
A living specimen is placed in a box to be examined with a microscope.
Prevent focus instability
The specimen must be kept alive so the temperature, CO₂ and O₂ levels are monitored and maintained.
What problems may arise from using microscopes to examine specimens for different timescales
- When only measuring for seconds, there may be problems with artifacts in multichannel/4d imaging
- when measuring from hours to days there may be problems with the viability of the specimen, also the positioning of the capture may change
Describe the triangle of frustration for signal detection
We can satisfy 2 of the three parameters when doing a measurement:
- Temporal resolution 1
- spatial resolution 2
- sensitivity 3
All detections have their benefits and limitations, what is best depends on the application requirments
What is spatial resolution
The size of the smallest unit of an image capable of distinguishing objects.
Where the bigger the pixel area the smaller the resolution and vice versa
Takes more time to capture an image with more resolution
What is Intensity resolution?
It refers to the number of intensity levels used to represent the image.
Where a higher bit value means larger intensity levels. (Shades of grey)
Takes more time to capture an image with more intensity
What are the markings on objective lens’
- Magnification
- Application
- Coverslip Thickness
- Numerical aperture/ Immersion medium
- Working distance
How does aperture determine resolution
- The higher the numerical aperture the better the resolution power of the objective.
RESOLUTION ≠ MAGNIFICATION
How is the function of light and lenses in a light microscope
To illuminate and magnify
How can light microscope be used to study cell histology
- look for proteins of interest using antibodies
How can light microscope be used to study cell morphology
- look for intact and denatured collagen
How can light microscope be used to study cell histology
- study the differentiation of cardiomyocyte-like cells derived from adipocytes
- study cell migration
What are the parts involved in electron microscopy
- Electron source
- Electron beam
- specimen
- Electromagnetic lens
- Viewing screen