| Project/Area Number |
59870005
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
General physiology
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
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| Project Period (FY) |
1984 – 1985
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| Project Status |
Completed (Fiscal Year 1985)
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| Budget Amount *help |
¥10,100,000 (Direct Cost: ¥10,100,000)
Fiscal Year 1985: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1984: ¥6,800,000 (Direct Cost: ¥6,800,000)
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| Keywords | dichroic mirror / two-wavelength optical recording system / membrane potential |
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| Research Abstract |
Using an Olympus Vanox Microscope with a fluorescence vertical illuminator attachment, we have constructed a two-wavelength optical recording system to simultaneously monitor changes in membrane potential and cellular events coupled with the membrane potential. This apparatus was designed and constructed to be able to make the following procedure. Light from a tungsten-halogen filament lamp was collimated, and focused on a preparation on the stage by means of the brightfield condencer on the microscope. The preparations were stained with potential-sensitive dyes and other molecular probes (as required). The light from the preparation was separated in two components by a dichroic mirror, on corresponding wavelengths. Two photodetectors were placed at the planes where the microscope objective makes magnified images in the upward (straight) and rectangular directions. Usually, the membrane potential-related absorption or fluorescence signals were detected in the upward (straight) direction and the changes in the membrane potential-coupled light scattering or absorption were monitored in the rectangular direction. In order to select an optimal wavelength, an interference filter was inserted in the front of the detector in each direction. The output of each detector was fed to an amplifier via a current-to-voltage converter, and then the amplified outputs were recorded simultaneously on a data recording system. We hope that the method reported here would enhance our understanding of the cellular basis of various physiological functions.
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