4. Set microscope to high magnification
Procedure
- On the microscope, select a higher power objective that will be suitable for viewing the objects of interest (e.g. tubules, nerve fibers, small blood vessels, microvilli, etc.).
- Select the corresponding lens from the Select High Mag Lens drop-down menu.
Recommendations
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We recommend using 60x, 63x or 100x oil objectives for counting. Such objectives provide the thin depth of field required to count accurately, especially when fibers are on top of each other.
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In general, lower power objectives, such as 10x, 20x, or 40x air objectives, do not have the adequately thin depth of field that is needed to distinguish the one fiber from another in areas where they are densely packed.
The item to be counted is called a “profile;” it corresponds to the point where the fiber transects the sphere boundary. If the fibers are thick, you must be able to identify the center point of the fiber and only count locations where the center point transects the sphere outline.