Question

Cell Biology: Microscopes

1) If you could use any color in the spectrum of visible light to observe your specimens in the microscope, what color would you use to obtain the highest resolution? Explain.

2) Nmae 3 differences between upright and inverted compound microscopes.

3) In what circumstances would you use an upright, an inverted and a fluorence microscope?

4) Would you use the same filter cube for every fluorophore you use in your experiment?

Answer 1

1) For high resolution detailing of the specimen, the color with the shortest effective imaging wavelength is required. Therefore near UV light is given first preference, then comes the blue in the visible light spectra, followed by green and red.

2) Upright Microscope:

• The objective is placed above the stage.
• Suitable for analyzing fixed samples.
• The light is directed on the specimen from below.

Inverted Microscope:

• The objective is placed below the stage.
• Suitable for live-cell imaging.
• The light is directed on the specimen from above.

3)

• An Upright microscope is useful for fixed sample visualization with more resolution.
• The inverted microscope is useful for live imaging.
• A fluorescence microscope is useful for visualizing specimens with a fluorophore, it uses reflected light from the image.

4) No, we have to use different filter cubes for different fluorophores. It mainly depends on the excitation and emission wavelength of the fluorophore.