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Room-Temperature Detection of a Single Molecule's Absorption by Photothermal Contrast A. Gaiduk, et al. Science 330, 353 (2010); DOI: 10.

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M. Mater.org on July 19. Zeng et al. The fluorescent state is easily destroyed or quenched by photochemical reactions. 6. S. Orrit* So far. M. Rev. 32 (1999). Y. which receives partial support from NSF through the National Nanotechnology Infrastructure Network program. E Stat. P. 2011 . Interdiscip. H. R. The integrity of sample SISO-3 after 24 hours of annealing under vacuum at 140°C was verified by size exclusion chromatography (SEC). ed. Winn. Rosen et al. B 59. virtually no bleaching event was observed. F. W. D.1126/science. 35. 99. *To whom correspondence should be addressed. McTague. Anderson. Diamant. Trans. X.1195552 Room-Temperature Detection of a Single Molecule’s Absorption by Photothermal Contrast A. Dupont de Nemours & Company. M. composition. However. 18. J. 43. Elmer. Dickins.T.. S1 to S7 References 23 July 2010. Meng et al. Phys. Blech. 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This photothermal technique provides contrast for the absorbing objects only. Denton. Photonic Crystals: Molding the Flow of Light (Princeton Univ. Macromolecules 32. Portions of this work were performed at the DuPontNorthwestern-Dow Collaborative Access Team (DND-CAT) located at Sector 5 of the Advanced Photon Source (APS). and the State of Illinois. for example. van Megen. DND-CAT is supported by E. Rev. This work was supported by the Department of Energy through a subcontract to UT-Battelle (4000041622). 22. J. On the basis of bleaching steps. Lodge. Matsushita. Phys. a modest 80 kg/mol). A. governed by wellestablished molecular parameters. J. 30.3 mm. 340 (1986). under which the absorption cross section of the narrowest molecular transition was extremely large. P. Schulz. Phys. Matsen. Seddon. 195502 (2007). should result in a unit cell dimension of a ≈ 0. X. 2 and 3) and mechanical spectroscopy (fig. Rev. 5. V.S. consistent with these simulations. potentially suitable for application as a photonic crystal (35). under contract DE-AC02-06CH11357. S. Lett. W.sciencemag. Nano Lett. New York. J. Mag. V. 3) because of the low relaxation to the ground state or to intermediate dark states. Macromolecules 43. D. the average bleaching time was of the order of 1 minute. Clark. For the past 20 years. 1602 (1980). A. Rev. 26. Parts of this work were carried out in the University of Minnesota I. Am. only a very small fraction of all absorbing molecules (called chromophores) are strongly fluorescent (then called fluorophores). E. Chapuis. Levine. J. 20. Supporting Online Material www. Berney. H. H. Macromolecules 13. CA. D. The s phase is the approximant crystal structure to certain dodecagonal quasicrystals. 5364 (1988).. T. we obtained an average absorption cross section of 4 angstroms2 for a single chromophore. S2) experiments with IL-15 may reflect a transient quasicrystalline morphology. J. Solid State Physics (Brooks/Cole. A. irrespective of scattering by defects or roughness. 7641 (1996). V. 464 (2007). Ashcroft. Indeed. was achieved by absorption (4) but relied on favorable cryogenic conditions. with lattice parameters that are expected to scale with the two-thirds power of block copolymer molecular weight. V. M. Alexander. 702 (1978). Macromolecules 15. A. Philos. S. Zeng. 57 (1979). 1208 (2003). 1 (1990). J. D. Netherlands. Department of Energy. G. S. C. The authors gratefully acknowledge helpful discussions with N. 9. Frank. 12. ed. P. Nature 320. C. Acta Crystallogr. Molecular simulations indicate that dodecagonal quasicrystals represent slowly evolving metastable states relative to the (equilibrium) s phase (18). Use of the APS was supported by the U. 33.. 15. B. Schneider. J. More generally. B 44. 1951 (1984). Kasper. J. S. T. A. Zhang. and subject to judicious choice of block types (which controls c). S. 27. Lifshitz. the National Science Foundation through grant DMR0704192. Bates. Bates.sciencemag. S. T. 745 (1997). T. Roth. often involving electron or proton transfer. G. Takano. Phys. with a signalto-noise ratio of ~10 for a single molecule in an integration time of 300 milliseconds. 2008). 36. Birkedal. Quasicrystals: A Primer (Oxford Univ. Acad. Acta Crystallogr. 9.e. The first single-molecule optical detection experiment. Meade. Acta Crystallogr. Sci. The s phase has an enormous unit cell. Basic Energy Sciences. Ruijgrok. Proc. Identification of the s phase in linear block copolymer melts presents the opportunity for designing interesting and useful materials from many other polymers. Hayashida. single-molecule imaging has predominantly relied on fluorescence detection. Lawson et al. Bates. 16.4 nets (25). B. Acta Crystallogr. monitoring optical absorption more directly would have definite advantages. Pusey. 95 (2005). We imaged single nonfluorescent azo dye molecules in room-temperature glycerol by the refractive effect of the heat that they release in their environment upon intense illumination. X. Topics 61. W. Dotera. 1994). F. N. Zeng. 167 (1999). 87. C. I. Decades of experimental experience and well-established theory show that single-component block copolymer melts exhibit universal phase behavior. 53. D. 2. Press. S. L. 131. 1976). 53. Werner. 4449 (2010). 324 (2003). and the University of Minnesota Materials Research Science and Engineering Center (MRSEC). Plasmas Fluids Relat. 297 (1956). C. S. Acta 1031. Douglas. Appropriately designed block copolymers could represent ideal materials with which to characterize the thermodynamic and kinetic properties of these fascinating aperiodic systems. C. Mag. Thus. S. J. Metall. D. M. by Kador and Moerner. M. Rev. 17. Y. W. S. 12. F. Press. 6. Their doubly modulated absorption technique was later improved on with modern optics and detectors (5–7). Mater.. C. Specht. Our time-dependent SAXS (Figs. the Dow Chemical Company. J. Ungar. F. B. Soft Matter 1. optical single-molecule detection has relied on fluorescence (2. Phys. Taylor.org/cgi/content/full/330/6002/349/DC1 Figs. Morse. Rheol. 26). 157 (2004). M. A. Such molecules could become more-natural labels than fluorophores. even after more than 10 minutes of illumination.A. Joannopoulos. Metall. F. 38. 3011 (2007). D. P. accepted 3 September 2010 10. DNA bases. Hurd. Steinhardt. J. and segmentsegment interaction parameter c (8). Bates. U. J. A 46. S. 6845 (2000). M. No blinking was observed in the absorption signal. M. W. 2. Characterization Facility. 87. N. Lett. 2300 RA Leiden. Natl. 24. In a solution saturated with oxygen. 8. W. 34. Gruner. Palmer. Phys. 25. J. Mermin. M. 14. F. Belmont.. Science 299. 235503 (2007). A. Janot. 28. Pattison. NJ. Percec.REPORTS component block copolymer melts. S. Schmidt. N. J. K. 3021 (2007). 85. Ungar. B. N. E-mail: orrit@molphys. J. Acta Crystallogr. Office of Science. G. A Phys. 89 (1988). Liu. Phys. primarily the molecular weight. 32. 4.org 15 OCTOBER 2010 353 Downloaded from www. 589 (1982). In the absence of oxygen. R. L. 41.sciencemag. Gratias. Soc. R2984 (1992). References and Notes 1. Keys. Ackerson. P. Today 52. 3. Gaiduk. 23. Arakcheeva. I. G. D. 19. N. M. Balsara and T. A. 29. Lodge. Nature 428. Macromolecules 29. Leiden University. Lett. R. Cohen. D. Fluorescence requires an efficient pathway for populating the emitting state and also the near absence of nonradiative SCIENCE VOL 330 www. an asymmetric diblock copolymer prepared with 20 times the molecular weight of IL-15 (i. Biochim. 21. Leibler. (6 Pt B). 2477 (1984). D. Shechtman. ingle-molecule optical detection (1) has become an indispensable tool in molecular biology and materials science. 11. Hillmyer. Phys. R. Glotzer. 2. C. Bates. J. Sci. S. B 53. Lett. 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however.01 1 probe power (mW) 1 10 100 -1 0 1 0.6 1.)) 1 A 2µm C z x D 500nm 0 100 500nm 0 photothermal (a. small particles (13).8 2. essentially because of fast dephasing by thermal fluctuations. (10.u. assumed to be spherical: sabs is the absorption cross section of the absorber.) heating power (mW) Fig. S1 (17). and laser noise. The number of the absorption events varies as the ratio of the absorption cross section to the area of a diffraction-limited light beam. 14) to detect some tens of molecules (12).1 0.1 0. The FWHMs of the photothermal spots are 210 nm and 370 nm. that is.org on July 19. Three main routes have been pursued in the past 10 years toward surmounting these obstacles.REPORTS absorption signal still appears on a strong background of unabsorbed photons. The backward scattered probe light is collected by the focusing microscope objective. sy = 250 nm. For most of the data. Photon noise on the probe beam is therefore considerably reduced. they were able to image absorption in cells down to a few tens of molecules. Boyer et al. demonstrating a signal-to-noise ratio of about 10 with reasonable integration times of 100 to 300 ms. for practical applications. (A) A raster scan image of a sample containing 10-nm (yellow) and 5-nm (red) diameter gold particles in glycerol: heating power is 0. Our setup for photothermal contrast imaging is shown in fig. Photothermal images of gold nanospheres illustrate the large dynamic range of the method and its low background. Solid lines show linear fits at low heating and probe powers. thereby slightly changing the detected probe intensity. probe power. and R the radius of the heat source. Because the probe wavelength can be chosen in a transparency region. Most recently.43 mW. a change in transmitted signal of 10−7 requires more than 1014 photons to be detected. in conditions under which many diffusing molecules contributed in turn to the signal. These data are reproduced from (18). VOL 330 SCIENCE www. also observed in confocal fluorescence microscopy. Second. because stimulated photons can only arise from the excited state of molecules.u.0 E 20 # 10nm 10 0. A method for the detection of singlemolecule absorption at room temperature must thus solve two different problems. We very recently used this setup (18) to improve the photothermal detection approach of y x log10 (photothermal (a. Note the two weak lobes of the PSF in the x.u. is rather low. it is vital to be able to discriminate true absorption from scattering. The local inhomogeneity of the refractive index scatters a probe beam. The photothermal signal is proportional to both the heating and probe powers. Because statisticalffiffiffiffi p fluctuations in a number N of photons scale as N . This ratio is about 10−7 for a laser spot diameter of 300 nm. various sensitive optical techniques have been applied either in interferometers (12) or in a reflection or transmission geometry (13. A disadvantage of the method is that the transduction efficiency.) photothermal (a. relating the index change to the dissipated heat. The detection of such small signals therefore requires many absorption events. Therefore. a. In the latter two cases.. 2011 . Error bars indicate standard deviation values. First. joined to excellent rejection of the scattering background. Of those. However. Iheat the pump intensity.u. (9) applied a similar method to detect single immobilized gold nanoparticles down to 5 nm in diameter. 3 ms. With a modulated stimulating beam. Detecting the absorption of a single molecule at ambient conditions is much more challenging than at low temperature because of the reduction by 5 to 6 orders of magnitude of the cross section. electronic. arising for instance from roughness of the interfaces. the absorption cross section of a typical chromophore at room temperature is only of the order of 10−2 nm2. (B) Histogram of the photothermal signals for 94 gold particles of 5-nm or 10-nm diameter. which may conflict with the poor photostability of many fluorescent molecules at ambient conditions. this inefficiency is more than compensated for by a gain in applicable probe intensity and the associated reduction in photon noise. The absorbed pump power gives rise to a temperature gradient with abs Iheat an amplitude DTsurf ¼ s4pkR at the surface of the source.01 1E-3 0. 11) and has led to a number of applications in recent years. Indeed. Xie and co-workers have explored stimulated emission to distinguish absorption from scattering (16). integration time per pixel.1 10 log10 (photothermal) (a. Photothermal contrast relies on the intensity change of a probe beam caused by a modulated heating beam of a different color. We pushed the sensitivity of photothermal detection to the single-molecule limit. 21 mW. this method efficiently rejects scattering and fulfills the second requirement. sent to a fast photodiode.sciencemag. the photothermal signal roughly scales as the volume of the particles. We have chosen the reflection geometry with index matching to adapt the probe signal to the characteristics of our fast low-noise detector (maximum detected intensity 18 mW). and fed into the lock-in amplifier. (Inset) Histogram of signals for 88 gold particles of 20-nm diameter in the same conditions. This inhomogeneous refractive index profile results from the heat dissipated by a pointlike absorber in its surroundings. As expected. (E) Linear scaling of the photothermal signal of a 20-nm gold particle with heating and probe (inset) powers. k the thermal conductivity of the medium. The resulting weak scattered field interferes with the main probe field (either transmitted or reflected). much higher than the saturation intensity of the heating beam. a molecule’s absorption appears on a background of scattering by any inhomogeneities in refractive index. the probe intensity can be very high. of the order of 10−4 K−1 or lower. and sz = 670 nm. a spectral range where absorption is negligible.) 100 10 1 1 30 B 5nm 30 15 0 20nm 1. Alternatively.org 354 15 OCTOBER 2010 Downloaded from www. as mapped by scanning a 20-nm-diameter gold particle in three dimensions.sciencemag. It is proportional to ∂n/∂T. Because a good overlap of the two tightly focused beams with different colors is critical. (C and D) The point-spread function (PSF) of the photothermal signal is well fitted by a Gaussian with sx = 220 nm. the use of ultraclean and stable conditions was essential to ensure the absence of scattering background. Photothermal contrast relies on a timedependent thermal lens. or quantum dots (15). resulting in a lower signal-to-noise ratio than fluorescence. 1. a performance that was later substantially improved by Berciaud et al.u. We modulate the pump at rather high frequency (around 1 MHz) to reject mechanical. at least 107 have to be absorbed by the molecule. z image. they are smaller than the symbol size. residual chromatic aberrations of the objective must be carefully compensated. arbitrary units. in a direct extinction measurement. where only missing photons are identified. Kitamori and co-workers (8) have detected sub-yoctomole concentrations of absorbers in solution with integration times of several seconds.

This value. we measured the photothermal detection volume by scanning a 20-nm gold sphere in three dimensions. A photothermal image obtained on a sample of spin-coated quencher-DNA construct (BHQ110T-BHQ1) submerged in nitrogen-bubbled glycerol is shown in Fig. 2. we demonstrated the detection of a dissipated power as low as 3 nW with a signal-to-noise ratio of SNR = 8 and an integration time of 10 ms. (10. Several typical background-corrected photothermal traces are displayed in Fig. The elongated shape of the photothermal spots in this image (210 nm and 370 nm principal half-maximum widths) arose from the shape of the pump beam.sciencemag. 2D. (B) Photothermal time traces showing single-step bleaching obtained on a sample with BHQ1-10T-BHQ1 on glass in oxygen-saturated glycerol: integration time per point. 1. we verified the linear relationships between pump and probe powers and photothermal signal by varying the heating power over 4 orders of magnitude (1 mW to 10 mW) and the probe power over 2 orders of magnitude (1 mW to 100 mW). C and D.1 Å2. Compared with our prior report (18). and 670 nm. sBHQ = 2. Less than 2% of the molecules bleached in the first 10 min of illumination. In our previous work (18). we only need to compare the smallest detectable power over a given integration time to the power dissipated by a single molecule. showing a huge reduction in the bleaching efficiency per photon compared with usual fluorophores in the same conditions. 5. 2C with a steplike cutoff at 3 sBHQ ). www. Stronger spatial filtering yielded a much better shape. We first characterized the sensitivity of our setup with gold nanoparticles used as absorption standards.05 nm² (iii) 6 4 D # (iv) (v) 0 20 40 2 0 1 2 3 time (s) log10(tstep(s)) Fig.08 0.3 Å2. (D) Histogram of survival times before one-step photobleaching events for the same set of data.04 0. 11). S2 and table S1) (17). The highest dissipated power at saturation of 15 nW well above the limit of 3 nW was obtained for a highefficiency dark quencher (Black-Hole-Quencher. The most favorable molecules for photothermal detection should therefore have strong nonradiative channels and thus very weak fluorescence yields. (C) Histogram of amplitude of 30 bleaching steps in the photothermal traces. the larger is the dissipated power at saturation. y. In (18). 2C summarizes the absorption cross-section changes in 30 single bleaching steps. 100 ms.1 mW. Note that the maximum value of the cross section for a favorably oriented molecule is 3sBHQ = 6. 250 nm. even under the heavy illumination to which we subjected them. we chose a chromophoreDNA construct to increase the absorption signal by the presence of two chromophores in each construct and to secure the adhesion of the constructs to the glass surface by their single-strand DNA linker (figs. Figure 1A shows an image of a sample containing 10-nm. The full widths at half maximum (FWHM. BHQ1. Slight deviations from linear behavior at high power may be due to variations of spectra or thermal constants of the materials in the broad range of temperature changes explored (from 0. respectively.1 mW at 514 nm. The signals from different diameters differ by a factor of about 8. When the glycerol was saturated by oxygen. because spatial filtering was weak in this experiment.02 0. 2B and demonstrate digital irreversible steps. 300 ms. The histogram in Fig.05 K up to about 500 K). determined by a Gaussian fit) across x. We have attempted measurements of the polarization dependence of single-molecule absorption signals. probe. The histogram of survival times before digital photobleaching is shown in Fig. We calculated the dissipated power at saturation for several commercially available organic molecules (fig. For the present experiments. heating power.06 0. This is the case for the trial molecules chosen in the present work.org SCIENCE VOL 330 nia). 2011 SNR . The image reveals four photothermal spots with a SNR ~ 10 for an integration time of 300 ms.and 5-nm-diameter gold spheres on glass surface in glycerol. Last. is in satisfactory agreement with the isotropic value. 20). The cutoff at 6. This azo dye molecule is used in the design of oligonucleotide probes to quench the fluorescence of a dye in a DNA construct (19. 2C is a broad distribution without distinct maximum. and z are 220 nm. 22) cross section of these gold nanospheres is 460 nm2 at 514 nm in glycerol. in good agreement with absolute cross-section measurements (23).1 Å2. as expected from the volume ratio. These spots persisted for as long as 1 hour without bleaching or fading. we exploit the thermal sensitivity of glycerol here but not the thermal isolation from the glass substrate. Among the different points discussed in (18). 84 mW. The red line represents the distribution of absorption cross sections of single chromophores corresponding to the isotropic distribution of transition dipole moments. 5. These data are reproduced in Fig. By increasing the integration time to hundreds of ms. but these 15 OCTOBER 2010 355 Downloaded from www. which corresponds to the cross section of a single chromophore.1 Å2. The histogram of Fig. This robustness can be attributed to the much shorter dwell time in the excited singlet state resulting from the efficient internal conversion. even smaller dissipated powers can be measured.sciencemag. 84 mW at 800 nm.REPORTS Berciaud et al. deduced from the absorption spectrum of the quencherDNA construct in glycerol. 2A. The absolute value of the change in absorption cross section upon photobleaching is estimated by comparison to the photothermal signal of 20nm gold nanoparticles.00 0. in qualitative agreement with the isotropic distribution of transition dipole moments (solid line in Fig.3 Å2 is the calculated cross section for a perfectly oriented molecule. and that the molecules are fixed to the surface and/or to the DNA linker. one-step photobleaching was observed after an average time of ~1 min. The calculated (21. The shorter the fluorescence lifetime (t F) and the lower the fluorescence quantum yield (hF) are. CaliforN N NO2 N N OCH3 N O oligo A 15 4 3 2µm 5 C # 2 1 0. S3 and S4) (17). consisting of two fluorescence quenchers (BHQ1) attached to a single strand of DNA (10 thymine bases): integration time per pixel. The average change in absorption cross section upon bleaching is 4. We measured photothermal signals by heating at 514 nm and probing at 800 nm. Biosearch Technologies. heating. Novato. The average time is 49 s. (A) Photothermal image of four constructs (BHQ1-10T-BHQ1). We never observed any blinking or any later reappearance of a bleached photothermal signal. The average number of absorbed photons before bleaching is about 1011 under oxygen-saturated conditions and larger than 1012 under oxygen-free conditions.org on July 19. with an average value of 4.10 B (i) (ii) σabs (nm 2 ) 0. in contrast. probe power. A single molecule can release heat along two pathways: a direct nonradiative transition from the excited state and vibronic relaxation before and after a radiative fluorescent transition. To ascertain the possibility of detecting a single organic molecule in photothermal microscopy. Our observation suggests that the absorption dipoles of BHQ1 molecules are not freely rotating. the choice of the transducing fluid is crucial to enhance the sensitivity.

S6 to S8) (17)]. which account for 25% of the total terrestrial greenhouse effect. B. Kador. Our successful single molecule detection relied here on favorable conditions. Sawada. Kukura. S1 to S9 References 22 July 2010. N2O. Renn. Orrit. L. Express 16. Boyer. Phys. D. Ed. M. Radiative forcing experiments assuming doubled CO2 and a 2% increase in solar irradiance (5) show that water vapor provides the strongest climate feedback of any of the atmospheric GHGs. Science 297. S. 50. Chem. A. plunging the global climate into an icebound Earth state. B. A. D. 033601 (2007). 24. Stahl. Phys. A. Phys. and that the continuing increase in CO2 due to human activity is therefore not relevant to climate change. J. An intrinsic limitation of the photothermal method is the low transduction factor between pump and probe because of the relatively weak variation of refractive index with temperature. 2112 (2001). does not condense and precipitate from the atmosphere at current climate temperatures.1 mW focused into the diffraction-limited spot and with an even higher probe power of more than 70 mW. Maali. Lett. 15. Rev. 3. 6. Sandoghdar. Crenshaw. B 73. C. Reto A. S9) (17). Hwang. 2880 Broadway. Arbouet et al. M. NY 10025. S. S. 1670 (1999). Totowa. M. for example. 80. This is because CO2. 7. These studies established long ago that water vapor and CO2 are indeed the principal terrestrial GHGs. Phys. 1. *To whom correspondence should be addressed. Rev. Gaub. 96. Rev. Gaiduk. 12. Wrigge. which was first discovered by Joseph Fourier in 1824 (2). P. 9. 73. 2006). Moerner. Renn. 021802 (2006). E. M. Marras. We only found very few examples of two-step photobleaching of the BHQ110T-BHQ1 constructs. X. Ruedy Ample physical evidence shows that carbon dioxide (CO2) is the single most important climate-relevant greenhouse gas in Earth’s atmosphere. If true. Interesting candidates are natural absorbers. M.and nano-optomechanical systems (25). Chem. joined with the low probability of finding two favorable orientations in the same construct. E. 1998). J.. S. Cognet. 111. M. W. Yorulmaz. CH4. Sandoghdar. further heating the ground and maintaining the temperature gradient in the atmosphere. Tokeshi. TE is also the Planck radiation equivalent of the 240 W/m2 of global mean solar radiation absorbed by Earth. 8. A. Lett. 073702 (2009). I. and precipitating. from which it is re-emitted in both upward and downward directions. further consideration shows that CO2 is the one that controls climate change.. 17358 (2008). P. R. We acknowledge financial support by European Research Council (Advanced Grant SiMoSoMa). 25. Lacis. Lett. 18. Gerhardt et al. then absorbed locally within the atmosphere. J. 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This identifies water vapor and clouds as the fast feedback processes in the climate system. A. 257402 (2004). Tamarat.* Gavin A. 5. Rev. Orrit. G. F. E-mail: andrew. Rev. Chem. Kador. We believe that this absence results from the low probability of bleaching. 22. Berciaud. A. Celebrano. 1160 (2002). Holtom. and showed quick correlated decays of both signals upon illumination (fig. Lett. 9. 127401 (2004). Such simultaneous signals arose only from abnormally bright spots. Lett. E. References and Notes 1. Sci. photomechanical or photoelectrical detections using micro. Fejer. 65. On the sample of Fig. 045424 (2006). H. e122 (2002). B. 2011 . Wild. S. 93. accepted 16 September 2010 10. Phys. Min et al. Possible interpretations are aggregates of several molecules desorbing simultaneously or. Lett. 17. 26. M. for example. G. Sci. W. Besides direct solar heating of the ground. P. Spectrosc. 3486 (2007). Lett. Without the radiative forcing supplied by CO2 and the other noncondensing greenhouse gases. 2535 (1989). Kitamori. Puchner. VOL 330 SCIENCE The Sun is the source of energy that heats Earth. S. NJ. Hibara. Didenko. New York.lacis@nasa.REPORTS measurements were very difficult because of the buildup of a photothermal background over long illumination times by the pump beam [the probe had no noticeable effect on this buildup (figs. Materials and methods are available as supporting material on Science Online. R. Lounis. Phys. Methods in Molecular Biology: Fluorescent Energy Transfer Nucleic Acid Probes: Designs and Protocols (Humana. Gehardt. are highly active components of the climate system that respond rapidly to changes in temperature and air pressure by evaporating. 11. Noncondensing greenhouse gases. S. R. Instrum. this result opens the way for further optimization of the technique and to a much broader variety of absorbing molecules. Y. G. CO2 is a well-mixed gas that does not condense or precipitate from the atmosphere. 62. 2. W. Ignatovich.org/cgi/content/full/330/6002/353/DC1 Materials and Methods Figs.org Downloaded from www. 8. David Rind. Lounis. with a high heating power of 5. Keller et al. the use of glycerol (with large ∂n/∂Tand poor heat conduction) instead of water. possibly with excited-state absorption of the intense probe beam. W. L. there is also indirect longwave (LW) warming arising from the thermal radiation that is emitted by the ground. 2. another chemical species with a large absorption cross section. U. Min. experimentally verified by John Tyndall in 1863 (3). Novotny. which shows one-step bleaching. Lounis. T. 21. Butter. Lu. G. Tyagi.1126/science. 1105 (2009). W. this would imply that changes in atmospheric CO2 are not important influences on the natural greenhouse I NASA Goddard Institute for Space Studies. Phys. Phys. 93. M. 23. and quantified by Svante Arrhenius in 1896 (4). A. 10. A. USA. 2716 (1990). Phys. A 73. Lasne. for example. G. Bohren. V. Phys. Appl. C. Orrit. Nature 461. 4. Appl.. we also observed a few spots with strong photothermal signals corresponding to an absorption cross section of about 1 nm2. Xie. Latychevskaia. 13. 96. Hecht. R. T. This misunderstanding is resolved through simple examination of the terrestrial greenhouse. 926 (2009). F. Ruijgrok. 19. M. A. which would be useful for applications in analytical biochemistry and medical assays (24). Moerner. Hwang. 12A (1996). the terrestrial greenhouse would collapse. condensing. whereas water vapor can and does.1195475 Atmospheric CO2: Principal Control Knob Governing Earth’s Temperature Andrew A. Moerner. Chong.org on July 19. We could not observe simultaneous fluorescence and photothermal signals from the spots of Fig. This work is a part of the research program of the Stichting voor Fundamenteel Onderzoek der Materie. van der Meer for precious help with the experimental setup. B. Huffman. Chem. Rev. E. Schmidt. A. attributable to large aggregates. Absorption and Scattering of Light by Small Particles (Wiley. W. Weder. V. on the other hand. Cognet. V. V. V. V. Blab. 8755 (1999). Blab. appears full of promise. P. Opt. The difference between the nominal global mean surface temperature (TS = 288 K) and the global mean effective temperature (TE = 255 K) is a common measure of the terrestrial greenhouse effect (GT = TS – TE = 33 K). R. 98. J. F. E.a. thus serve to provide the stable temperature structure that sustains the current levels of atmospheric water vapor and clouds via feedback processes that account for the remaining 75% of the greenhouse effect.. A. Orrit. Nano Lett. For example. J. 30.sciencemag. and chlorofluorocarbons. Bernard. M. Nucleic Acids Res. New York. Phys. This radiative interaction is the greenhouse effect. Supporting Online Material www. metal proteins such as hemoglobin.