Volume 22, number I

CHEMICAL PHYSICS L.ETTERS

1.5 Scptcmber

1973

DIMER Takashi KAJIWARA,

SPEC-IXA

OE; RHODAMINE

I3

Richard W. CHAMBERS * and David R. KEARNS

o/C/zemis~ry, Urrilvrsify ofCuli/cjrrrio,
LX4 Rivcrsidc, CaJi;Fomfa 92502,

Departmerll

Rcceivcd 31 May 1973

The visible sbvJrption fluorescenrr and phosphorescence spectra and the polarized fh~orcsccncc and phosphoresccnce cxcitation spectra hwc been measuied for the dimer form of rhodnmine B. The rhodamine B dimer is somelvhat unique in that it exhibits both fluorescence and phosphorescence wherus other dyes usually cshibit onIy phosphorescence from the dimer. This unusual siruntion hz permitted us to ~rry out the first dctailcd eudv oE the polxized absorption 2nd emission spectra of rhodnminc I3 dimers. Simple esciton theory, cor;~mor?ly ucd to interpret the spectral properties of dimers, cannot account ior the polarization properties ol the absorption and en~isti~~fi sp~ct~a of tlvz rhodaminc B dimer.

1. Introduction Rhodamine B is one of a number of common organic dyes which readily form dimer species in aqueous solution [l-l 31. One unique feature of rhodamine dimers is that they show fluorescence 3s well as phosphorescence whereas some other similar dyes such as acridine dyes, fluoresceins and eosins show only phosphorescence in the dimeric state, [I 3,141. The f7uorescence spectrum of rhodamine B is also somewhat unique in that it has well defined vibrational structure, which can provide useful information about the interaction of two constituent molecules. In the present study we have investigated the polarized emission and excitation spectra of rhodamine B and determined the relative polarization of visjble transitions of the dimer, As we shall show, simple cxciton theory in which vibronic interaction is neglected cannot account for the observed polarization properties. However, with the inclusion of vibronic effects and slight asymmetry in the dimer con!iguration, it is possible to account for the features seen in the absorption and fluorescence spectra of rhodamine B dimers.
* Pre.sent address: Aerospace Corp., El Segundo, California, LISA.

2. Materials and methods Rhodamine B (Chroma-Gesellschaft Schmidt Company) was used as received. Spectroscopic measurements were carried out using a previously described fluorophosphorimeter (Chambers and Reams) [ 131 which permitted us to measure both fiuorcscence and phosphorescence excitation spectra as well as emission spectra. Polarized excitation and emission spectra were obtained by incorporating ;1polarizing sheet on the output side of the exciting monochromator and a ~013. coat polarjzer (Polacoat Inc.) in front of the detector. A 180 configuration was used in fluorescence polarization experiment, but a 90 configuration had to be used for excitation polarization experiment to ciiminate interference from the exciting light in measi_trement of weak lowest-energy absorption region. In this 90 configuration the incident exciting light was nearly normal to the front face of a flat (i mm) ceil and the emission from the front face was obsemed at approximately 90 to the incident @ht. The poiarization of the exciting light was perpendicular to the plane of incidence?, and parallel to the flat front face
t The plane ofincidcna is a plzne that includes the incident and reflected light rays. See any text of optics, e.g., ref. 1151.

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CHEMIC~_LPHYSlCS LEITERS

15 September 1973

of the sample cell. This geometrical arrangement allows an almost equal probability for deteciir.g emitted light polarized parallel and perpendicular to the exciting light. The polarization ratio is affected by many factors, including how well the exciting light is collimated, what part of the emitted light is effectively detected, how great the effect of refraction and scattering is, and so on. Since we did not try tc cbtain the maximum polarization ratio, our measured value may be somewhat smaller than can be expected from WIIple theoretical calculation. For detection an RCA 7265 photomultiplier was used with appropriate fil-

ters fcx excitation spectra measurement and with a McPherson Model 218 scanning monochromator for
emission spectra mcaslrernent. A Cary Model 14 recording spectrophotorneter was used to measure the absorption spectra. All spectroscopic measurements were carried out at room temperature or at 77K. Dimers are easily formed at 77OK in water containing 10 M LiCl, which is added for formation of clear glass without cracks. In order to obtain monomer spectra t!ze dyes were dissolved in 90% ethanol - 10% methanol solution, 50% ethanol - 50% 10 hl LiCl-wzter solution or 3: 1 glycerol-water. These solutions formed clear glasses but usually suffered considerably from cracking. For absorption measurements a 1 cm rectangular Pyrex cell was used and a 1 mm thick 1 cm wide flat quxtz cell was used for emission and excitation spectra measurements.

Fig. 1. Tile 77K absorption spectra of rho&mine B dimers in 10 hl 1X1-wotdr (solid line) and monomers in 9: 1 ethanol: methanoi solution (broken line). Both solutions were approsimately 10S5 hi. Individual bands (numbered l-4) are dis-

cussed in the text.

3. Results

The absorption spectn;m of the Aodamine B disolution at liquid N, temperature is shown in fig. 1 along with the coriespondmer in 10 M LiCl-water ing monomer absorption spcc:ru,m. In the rhodamine B dimer spectrum, we observed a weak bhnd at 603 run, which is designated as band 1 in fig. l! in addition to three stronger bands (bands 2, 3 and 4 in fig. 1) in the higher energy regjon. The weak origin band is not due to impurities, since we have observed no corresponding absorption in monomer spectra of the same solution at room temperature OI in the monomer spectrum of the same dye in 3 : 1 glycergl-water solution at 77K. The polarized fiuorescence excitation spectra are shown jn fig. 2 toge:hzr with their polarization iatios. The low energy band of dimer iluo38 .... 1.. ,.

rescence (650-7 10 nm, designated as band 2 in the fluorescence spectrum shown in fig. 2) was monitored using a cut-off filter (Coming Cs-2-64j and an interference filt;:r transmitting at around 670 nm (Spectracoat Monopass Filter 3139-666, Optics Technology, Inc.). The light source was a 5OOW Xe-lamp combined with a Hilger-En@ model 1000 monochromator. The dimer fluorescence polarization spectra are shown also in fig. 2 together with their polarization ratio. The light source for fluorescence polarization measurements was a Bausch & Lomb mercury light source P-200. A Bausch & Lomb low resolution grating monochromator and an interference fi!ter (Spectracoat hIonopass Filter 3 139-533, Optics Technology, Inc.) were used to select the 5460.7 A mercury line to excite the strong absorption band of dimer molecules. From the fluorescence excitation spectrum and its polarizatiori ratios, we conclude that the polarization directions of all strong absorption bands (+2, 3 and 4) are parallel to band 2 in fluorescence, and that the polarization direction of the weak origin band (band 1) is oblique to it. Fluorescence polarization ratios further confirm that the strong absorption bands are po. larized perallel to the band 2 in fluorescence. It is also obvious that the origin (band 1) of fluorescence is

oblique!y polarized to strong absorption bands 2, 3 and 4. Because of strong interference from the exciting light we could not directly observe the polarization
of the origin absorption band 1 by monitoring the or-

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CHEMICAL

PHYSlCS

LETTERS

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1973

18

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Fig. 2. Fluorescence excitation and emission tion ratios are also presented in this figure. tored (650-710 nm). A part of the spectra ture at lower energy region. The small band hrization ratio is due to residual monomer tion ratio is due to dimcr phosphorcsccncc.

spectra of rhodaminc D dimcrs in 10 %I LiCI-water solutions at 77K. The poIxtiztIn the fluorescence excitation measurements the second tluorcwence band W;LSmonimwsured at higher sensitivity is also shown by broken lines to show the detailed SUUCwhich is observed at the high cncrgy side of the dimer fluorcsccnce and giver a high POfluorescsnce. The small shoulder on the lower energy side which gives 3 IOW pol?riza-

igin band 1 of fluorescence. However, it seems safe to coxclT.lde that they are polarized parallel to one another. Simple Davydov theory I16--191 has often been used to interpret the electronic spectra of dye dimers and the essential results of this theory are indicated in fig. 3. Here we see that dimer formation is predicted to give rise to a resonance splitting with transitions to the upper (q,) state being strongly allowed, and the transition to the lower energy (9_) state being much weaker and opposite!y polarized with respect to t,he higher energy transition. In previous studies of the rhodamine B dimer absorption [l] the weak origin band 1 was not observed, so the strong band 3 in the absorption spectrum was assigned as the transition to the @+ state and the weaker lower energy band 2 was assigned as the transition to the 9_ state. This assignment cannot be correct since our results demonstrate that these two transitions have the suvw polarization, whereas the simple Davydov theory requires opposite polarization. Furthermore, this leaves the weak origin band which we observe unassigned. Alternatively the origin band could be assigned as the transition to the q_ state and the higher energy and somewhat stronger band 2 could be assigned as the transition to

cl

Fig. 3. A schematic drawing illustrating the effect of dimcr formation on the spectroscopic properties a dye zccordtig to simple exciton theory. In the di.neric state (D) the rrx&tion to the upper state is strong and polarized prrpendicul;lr

of

IO the weaker.

lower

energy

transition.

the q~+ state. The difficulties with this assignment arc that it does not provide an assignment for the strongest band (ban3 3) in the spectrum, and furthermore band 1 is not polarized perpendicular to band 2, but rather obliquely io it. The complex polarization properties of the dimer fluorescence also indicate that there ore difficulties which are not easily understood in terms of the simple exciton model. It is evident from these cbservations Lhat the simple exciton model, as it is commonly applied !o the
39

Volume

22, number

CHEhIICAL PHYSICS LETTERS

15 September 1973

interpretation of dimer spectra, is inadequate. The problem with the sirnple exciton model is that vibronic interactions have been neglected and these must be included in order, to account for most of the spectral features of the dimer absorption spectra. In a subsequent paper we shal show that a resonance coupling theory which includes vibronic: interactions can provide a satisfactory account of t!le dimer absorption and emission spectra.

[4] K.L. Arvan and N.E. Zaitseva, Opt. i Spektroskopiya 11 (1961) 38. (51 G.P. Gurinovich and T.I. Stelkovn, Biofnika 8 (1963)
[6] [7] 229. L.V. Levshin and I.S. Lonskaya, 11 (1961) 148. L.V. Levshin and E.G. Baranova, Opt. Opt. Opt. i Spektroskopiya i Suektroskopiya

Acknowledgement

The support of the U.S. Public Health Service (Grant GM 10449 to DRK) is most grntefully acknowledged.

Izv. &ad. Nauk SSSR Ser. Fiz. 27 (1963) 591. [lOI L.V. Lwshin, Izv. Aknd. Nauk SSSR Ser. Fiz. 29 (1965) 1299. 1111 L.V. Levshin and D.M. Akbarova, Zh. Prikl. Spektroskopii 2 (1965) 43. LZI L.V. Levshin and D.Pl. Akbarova, Zh. Prikl. Spektro&OF& 3 (1965) 326. 1131 R.W. Chambers and D.R. Kearns. Photo&em. Photobiol.

6 (1959) 31. [81 L.V. Levshin and V.G. Bocharov, 10 (1961) 330. [91 V.G. Bocharov and L.V. Levshin,

i Spektroskopiya

10 (1969) 215.
Y.G. hior~zov, Biofizika 8 (1963) 388. Optics, translated by 0. Laporte and [I51 A. Sommerfeld, P.A. hfoldauer (Academic Press, New York, 1964) p. 13. 1161 AS. Davydov, Theory ol molecular excitons. translated [I41

References
[l] Th. Fijrster and E. Konig, 2. Eiectrochem. 61 (1957)
344.
[2]

by hl. Kasha and M. Oppenheimer Jr. (McGraw-Hill, New York, 1962). 1171 E.G. hlcRae and hl. Kasha, J. Chem. Phys. 28 (1958)
721.

L.V. Levshin and V.K. Gorshkov,

Opt.

i Spektroskopiya

1181 E.G. hlcRae

10 (1961) 401. [3] L-A: Ignateva, L.V. Levshin, T.D. Gripova and Y.M. Polukhin, Opt. i Spektroskopiya 13 (1962) 2i9.

and hi. Kasha, Physical processes in radintion biology (Academic Press, New York, 1964) p. 23. Pure iI91 hl. Kasha, H.R. Rawls and bl. Ashraf El-Bayoumi, Appl. Chem. 11 (1965) 371.