Design for supported planar waveguides for obtaining mid-IR evanescent-wave absorption spectra from biomembranes of individual cells

被引:18
作者
Braiman, MS
Plunkett, SE
机构
[1] Univ. Virginia Hlth. Sci. Ctr. #440, Department of Biochemistry, Charlottesville
关键词
FT-IR; Ge waveguide; IR microscope; theoretical sensitivity;
D O I
10.1366/0003702971940666
中图分类号
TH7 [仪器、仪表];
学科分类号
0804 ; 080401 ; 081102 ;
摘要
We have been developing miniature planar Ge waveguides to detect mid-IR evanescent-wave absorption spectra from the cell membranes of individual intact frog eggs, 1,5-mm in diameter, from Xenopus laevis, with the aim of detecting and analyzing transient conformational changes of voltage-gated ion channel proteins in the membrane, Here we use waveguide optical theory to calculate optimal dimensions for a germanium waveguide to be used as a multiple-internal-reflection ATR element for this purpose. We assume that light from a standard broad-band IR source Is coupled efficiently into and out of a planar Ge waveguide, and then onto a small-area MCT detector, by using an IR microscope with a high-numerical-aperture objective. To increase the coupling efficiency even further, we assume that we can fabricate the waveguide with a gradual 7-fold-tapering to the tiny dimensions needed in the sensing region. Then, assuming that similar to 10(7) ion channel proteins in an occyte can be made to contact an area of a planar Ge waveguide up to similar to 200 mu m in diameter, we calculate that voltage-gated structural changes in these channel proteins should produce absorbance change signals of similar to 10(-6) if the waveguide sensor thickness is set near the optimal thickness of similar to 1 mu m and the sensor region length is limited to 100 mu m. If such a sensor can be fabricated, we calculate that detection of the predicted voltage-gated absorbance changes with a commercial FT-IR microscope should be possible after similar to 20 min of signal averaging.
引用
收藏
页码:592 / 597
页数:6
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