Abstract

The construction and preliminary application of a thin-layer optical cell suitable for measuring absorption and fluorescence of highly absorbent solutions are described. The light path length is easily variable down to 2.5 μm with a commercial torque driver. Both absorption and fluorescence are measurable in a cell by conventional spectrophotometers and spectrofluorophotometers. The emission spectra of concentrated rhodamine B aqueous solutions up to 15 mM were measured with the thin-layer cell. The thin-layer cell showed great improvement in concentration quenching because of its very short light path length.

© 1984 Optical Society of America

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References

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  1. C. W. Robertson, D. Williams, J. Opt. Soc. Am. 61, 1316 (1971).
    [CrossRef]
  2. I. L. Tyler, G. Taylor, M. R. Querry, Appl. Opt. 17, 960 (1978).
    [CrossRef] [PubMed]
  3. G. Weber, F. W. J. Teale, Trans. Faraday Soc. 53, 640 (1958).
    [CrossRef]
  4. K. Horiuchi, H. Asai, Biochem. Biophys. Res. Commun. 97, 811 (1980).
    [CrossRef] [PubMed]

1980 (1)

K. Horiuchi, H. Asai, Biochem. Biophys. Res. Commun. 97, 811 (1980).
[CrossRef] [PubMed]

1978 (1)

1971 (1)

1958 (1)

G. Weber, F. W. J. Teale, Trans. Faraday Soc. 53, 640 (1958).
[CrossRef]

Asai, H.

K. Horiuchi, H. Asai, Biochem. Biophys. Res. Commun. 97, 811 (1980).
[CrossRef] [PubMed]

Horiuchi, K.

K. Horiuchi, H. Asai, Biochem. Biophys. Res. Commun. 97, 811 (1980).
[CrossRef] [PubMed]

Querry, M. R.

Robertson, C. W.

Taylor, G.

Teale, F. W. J.

G. Weber, F. W. J. Teale, Trans. Faraday Soc. 53, 640 (1958).
[CrossRef]

Tyler, I. L.

Weber, G.

G. Weber, F. W. J. Teale, Trans. Faraday Soc. 53, 640 (1958).
[CrossRef]

Williams, D.

Appl. Opt. (1)

Biochem. Biophys. Res. Commun. (1)

K. Horiuchi, H. Asai, Biochem. Biophys. Res. Commun. 97, 811 (1980).
[CrossRef] [PubMed]

J. Opt. Soc. Am. (1)

Trans. Faraday Soc. (1)

G. Weber, F. W. J. Teale, Trans. Faraday Soc. 53, 640 (1958).
[CrossRef]

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Figures (5)

Fig. 1
Fig. 1

Front view and cross-sectional view of the thin-layer cell. (a) Front view: B, brass block; b, bolt to tighten the brass blocks; and W, window. (b) Cross-sectional view at AA′ in (a): B, brass block (inside of brass block is flat within 10 μm); Q, quartz plate; and S, sample solution.

Fig. 2
Fig. 2

Sketch of the thin-layer cell in use. Cell holders for both the spectrophotometer and spectrofluorophotometer are not shown: (a) absorption measurement in the longitudinal position; (b) fluorescence measurement in the lateral position.

Fig. 3
Fig. 3

Thickness of the layer in the cell with the torque to tighten the brass blocks. 1.0 mM of rhodamine B aqueous solution was used to estimate the thickness from the absorbance at 554 nm.

Fig. 4
Fig. 4

Fluorescence emission spectra of rhodamine B aqueous solutions measured in a conventional 1-cm cell. Excitation wavelength was 355 nm. Slit widths of excitation and emission were both 5.0 nm. Concentration was 1:0.10 μM, 2:1.0 μM, 3:10 μM, 4:100 μM, 5:500 μM, 6:1.0 mM, and 7:5.0 mM. Fluorescence intensities at emission peaks except 7 were normalized to about unity with attenuation and amplification factors of 1.0, 1.2 × 10−1, 3.5 × 10−2, 6.0 × 10−3, 1.5 × 10−1, 1.0, and 2.0, respectively. Wavelengths at emission peaks were 574, 576, 585, 594, 612, 630, and 646 nm, respectively.

Fig. 5
Fig. 5

Fluorescence emission spectra of rhodamine B aqueous solutions measured with the thin-layer cell. Excitation wavelength and slit width are the same as in Fig. 4. The thickness of the layer in the cell was normalized to 3.0 μm estimated from the absorbance at 554 nm. Concentrations were 1:50 μM, 2:100 μM, 3:200 μM, 4:500 μM, 5:1.0 mM, 6:2.0 mM, 7:5.0 mM, 8:7.0 mM, 9:10 mM, and 10:15 mM, respectively.

Equations (2)

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C limit = OD E m · L = 1 E m × 1.6 × 10 4 ( M ) ,
L 0.01 = 0.01 2.2 × 10 4 · C = 4.5 × 10 - 7 × 1 C ( cm ) ,

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