Microscopy Flashcards
what are 5 basic essential parts of a microscope
Detector(PMT,CCD) Objective (+- immersion medium) Specimen (cover glass) Light conditioning system Light source (Halogen, XBO...)
what are the 3 things needed for the specimen
cover glass
sample surrounded by embedding medium
glass slide
what ensures that the temperature of specimen and microscope remain equilibrated and tightly controlled
a incubator box combined with a precision air heater
what is the experimental timescale of microtubule based movement
second, minutes
what is the experimental timescale of cytoskeleton
seconds, minutes, hours
what is the experimental timescale of cell motility
minutes, hours
what is the experimental timescale of differentiation
hours to days
what is the experimental timescale of development
hours to day
what are the 3 parts of ‘triangle of frustration’
Temporal resolution , spatial resolution , sensitivity
relationship with pixels and resolution
the smaller the pixel , the higher the resolution
what are the 6 marking on an objective
Magnification(i.e. 100X) Application (i.e. DIC H) Coverslip thickness mm working distance mm Numerical aperture/immersion medium
what is the aperture
the aperture of the objective determines the resolution. the higher the numerical aperture the better the resolution power of the objective
what does bleaching of fluorochrome mean
due to high intensity illumination the fluorophores might permanently loose their ability to emit light
How to avoid bleaching of fluorochrome
work with reduced excitation light intensities or grey filters
use shorter exposure times
use anti bleach in your mounting media
Advantage of confocal microscope
Higher Z-resolution and reduced out -of-focus blur make confocal pictures crisper and clearer
