| Simultaneous measurement of ciliary beat frequency and [Ca2+]i | ||
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To correlate changes in ciliary beat frequency and
[Ca2+]i it is important to measure each parameter
simultaneous. This is achieved by using different wavelengths of light
to monitor each process. |
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| Measurement of
ciliary activity White light from a halogen source is filtered at 645 nm to generate a red beam that illuminates the specimen and passes through the microscope and is reflected to the CCD camera. The red beam of 645 nm passes through the dichroic mirrors with cut-offs at 400 and 600 nm. The resulting high-speed images are recorded to an optical memory disc recorder (OMDR). |
Measurement of
[Ca2+]i The excitation light of 340 or 380 nm for the dye fura-2 is generated by filtration of a Hg arc lamp (cyan). This light is directed to the specimen by a 400 nm dichroic mirror. The resulting fluorescent light at 510 nm (green) passes through the first dichroic mirror but is reflected by the second, 600 nm dichroic mirror through a 510 nm filter to the silicone intensified camera (SIT). The images are recorded on a separate OMDR. |
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