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Qualifying the Effect of Microscope Properties and Techniques

Submitted by rmmcdonald on Thu, 10/03/2019 - 12:35

The title of the poster my lab partner and I created was, "Qualifying the Effect of Microscope Properties and Techniques". In a series of experiments we learned how to properly use a research quality light microscope. The experments were geared toward practicing the use of technical ascepts of the microscope. So this involoves the use of filters, shutters, and adjsuting numerical aperature. There were also a variety of methods we practice in order to capture a high quality image. These methods include setting up Kohler illumination, use of fluoroescent microscopy, and setting up an oil immersion lense. All these various experiments came together to encapsultate our main objective of qualifying the effect of microscope properties and techniques in order to create a high quality image.

A major part of our experiments was adjusting numerical aperature under a variety of different conditions in order to see its effect on the visualization of a sample. The following figure shows a change in numerical aperature when the magnification is kept constant  x100 and visualized with fluorescense. The fluorescent beads are figure A-C. The numerical aperature increases left to right: starting with 0.5, increasing to 0.875, and finally ending at 1.25. This is similar for the photos of the DAPI stained nuclei. Visually, we can conclude that both the DAPI stained nuclei and the fluorescent beads increase in image intensity as numerical aperature increases because we can see it gets brighter. This observation is quantified in the table because we mapped the fluorescent intensity of the five nuclei over the three different numerical aperature settings. The intensity of 1.25 is much greater than 0.5 proving that intensity increases as numerical aperature increases. It can also be argued that resolution increases too because the minimal loss of image light.