Exploring Single-Nanoparticle Dynamics at High Temperature by Optical Tweezers.

The experimental determination of the velocity of a colloidal nanoparticle (vNP) has recently became a hot topic. The thermal dependence of vNP is still left to be explored although it is a valuable source of information allowing, for instance, the discernment between ballistic and diffusive regimes. Optical tweezers (OTs) constitute a tool especially useful for the experimental determination of vNP although they have only been capable of determining it at room temperature. In this work, we demonstrate that it is possible to determine the temperature dependence of the diffusive velocity of a single colloidal nanoparticle by analyzing the temperature dependence of optical forces. The comparison between experimental results and theoretical predictions allowed us to discover the impact that the anomalous temperature dependence of water properties has on the dynamics of colloidal nanoparticles in this temperature range.

Nonlinear-Optical Response of Prussian Blue: Strong Three-Photon Absorption in the IR Region.

The nonlinear-optical properties of Prussian Blue nanoparticles have been evaluated with the use of femtosecond Z-scan measurements in the 1350-1750 nm range. This well-known inorganic pigment having interesting magnetic and electrochemical properties was found to be an efficient near-IR three-photon absorber. The maximum of the effective three-photon cross section is as high as 4.5 × 10-78 cm6 s2 at 1375 nm. By a comparison of the three-photon molar-mass-normalized merit factors, σ3/M, we show that this material is a competitive multiphoton absorber, especially in comparison to semiconductor quantum dots.

Uniting Ruthenium(II) and Platinum(II) Polypyridine Centers in Heteropolymetallic Complexes Giving Strong Two-Photon Absorption.

New trinuclear RuPt2 and heptanuclear RuPt6 complex salts are prepared by attaching Pt(II) 2,2':6',2"-terpyridine (tpy) moieties to Ru(II) 4,4':2',2":4",4"'-quaterpyridine (qpy) complexes. Characterization includes single crystal X-ray structures for both polymetallic species. The visible absorption bands are primarily due to Ru(II) → qpy metal-to-ligand charge-transfer (MLCT) transitions, according to time-dependent density functional theory (TD-DFT) calculations. These spectra change only slightly on Pt coordination, while the orange-red emission from the complexes shows corresponding small red-shifts, accompanied by decreases in intensity. Cubic molecular nonlinear optical behavior has been assessed by using Z-scan measurements. These reveal relatively high two-photon absorption (2PA) cross sections σ2, with maximal values of 301 GM at 834 nm (RuPt2) and 523 GM at 850 nm (RuPt6) when dissolved in methanol or acetone, respectively. Attaching Pt(II)(tpy) moieties triples or quadruples the 2PA activities when compared with the Ru(II)-based cores.

Beyond Single-Wavelength SHG Measurements: Spectrally-Resolved SHG Studies of Tetraphosphonate Ester Coordination Polymers.

Powder second-harmonic generation (SHG) efficiencies are usually measured at single wavelengths. In the present work, we provide a proof of concept of spectrally resolved powder SHG measured for a newly obtained series of three non-centrosymmetric coordination polymers (CPs). CPs are constructed from tetrahedral linker-tetraphenylmethane-based tetraphosphonate octaethyl ester and cobalt(II) ions of mixed, octahedral (Oh), and tetrahedral (Td), geometries and different sets of donors (CoO6 vs CoX3O). Isostructurality of the obtained materials allowed for the determination of anion-dependent tunability of SHG optical spectra and their relationship with solid-state absorption spectra.

Core/shell quantum dots encapsulated in biocompatible oil-core nanocarriers as two-photon fluorescent markers for bioimaging.

Highly fluorescent quantum dots (QDs)-loaded nanocapsules, intended for fluorescent cell imaging, were prepared via an emulsification/solvent-evaporation method. CdSe/ZnS core/shell quantum dots were applied as cargo; Poloxamer 403 as the polymer component; Cremophor EL as the nonionic surfactant; and mineral oil, oleic acid, or silicone oil were applied as the oil phases. Transmission electron microscopy, atomic force microscopy, dynamic light scattering, and zeta potential measurements were used to characterize the novel QDs-labeled nanoparticles by particle size, distribution, and morphology, as well as by ζ-potential and physical stability. The fabricated long-lasting nanocapsules exhibit good luminescence properties upon both one-photon and two-photon excitation. The potential of the encapsulated QDs for fluorescent imaging was evaluated in cytotoxicity studies as well as in imaging of intracellular localization, accumulation, and distribution of QDs delivered to well-characterized human cancer cell lines--doxorubicin-sensitive breast (MCF-7/WT) and alveolar basal epithelial (A549)--as well as on normal human umbilical vein endothelial (HUVEC) cells, as investigated by confocal laser scanning microscopy (CLSM). The colloidal CdSe/ZnS-loaded nanocapsules are shown to exhibit strong two-photon-induced luminescence upon excitation in the NIR optical transmission window spectral range, making them ideal markers for bioimaging application. The total two-photon cross section of a single nanocapsule was determined to be about 4.1 × 10(6) GM at 800 nm.

Two-photon excited luminescence of sulfur quantum dots for heavy metal ion detection.

Spectrally-resolved third-order nonlinear optical properties of water-dispersed sulfur quantum dots (SQDs) were investigated in the wavelength range from 740 nm to 820 nm with the two-photon excited emission technique using a tunable femtosecond laser system. The maximum value of the two-photon absorption (TPA) cross-section (σ2) for ∼5.4 nm size SQDs was found to be 185 GM (Goeppert-Mayer unit), while the two-photon brightness (σ2 × Î·) was found to be 1.5 GM at 780 nm, the wavelength being in the first biological transmittance window. The TPA properties are presented here as appropriate cross-sections normalized per molecular weight which enables meaningful comparison of the nonlinear factors of the studied quantum dots with those of various nanomaterials. The optimized TPA properties of these hydrophilic colloidal SQDs may be potentially useful for detection of Fe3+ metal ions. The experimentally determined limit of Fe3+ detection for both one- and two-photon regime was 10 µmol L-1 (0.6 µg mL-1). Förster resonance energy transfer between SQDs as donors and Fe3+ metal ions as acceptors was confirmed as one of the possible detection mechanisms using a time-correlated single photon counting technique.

Heptametallic, octupolar nonlinear optical chromophores with six ferrocenyl substituents.

New complexes with six ferrocenyl (Fc) groups connected to Zn(II) or Cd(II) tris(2,2'-bipyridyl) cores are described. A thorough characterisation of their BPh4(-) salts includes two single-crystal X-ray structures, highly unusual for such species with multiple, extended substituents. Intense, visible d(Fe(II))→π* metal-to-ligand charge-transfer (MLCT) bands accompany the π→π* intraligand charge-transfer absorptions in the near UV region. Each complex shows a single, fully reversible Fe(III/II) wave when probed electrochemically. Molecular quadratic nonlinear optical (NLO) responses are determined by using hyper-Rayleigh scattering and Stark spectroscopy. The latter gives static first hyperpolarisabilities ß0 reaching as high as approximately 10(-27)  esu and generally increasing with π-conjugation extension. Z-scan cubic NLO measurements reveal high two-photon absorption cross-sections σ2 of up to 5400 GM in one case. DFT calculations reproduce the π-conjugation dependence of ß0, and TD-DFT predicts three transitions close in energy contributing to the MLCT bands. The lowest energy transition has octupolar character, whereas the other two are degenerate and dipolar in nature.

Third-Order Nonlinear Optical Properties of Aqueous Silver Sulfide Quantum Dots.

Wide spectral wavelength range (500-1600 nm) measurements of nonlinear optical properties of silver sulfide (Ag2S, with 2- or 3-mercaptopropionic acid, 2 or 3MPA ligands) quantum dots (QDs) in aqueous colloidal solutions were performed using the Z-scan technique with tunable ∼55 fs laser pulses at 1 kHz. We have identified regions of the occurrence of various NLO effects including two-photon absorption, nonlinear refraction, as well as saturation of one-photon absorption. At the same time, we evaluated the relationship between the properties of the QDs and the variation of the material that covers their surface. The peak two-photon absorption cross section (σ2) values were determined to be 632 ± 271 GM (at 850 nm) for Ag2S-2MPA QDs and 772 ± 100 GM (at 875 nm) for Ag2S-3MPA QDs. The physicochemical factors influencing the three-dimensional self-organization of Ag2S QDs in water as well as their impact on spectroscopic properties were also investigated.

Difluoroborate-based bichromophores: Symmetry relaxation and two-photon absorption.

Given potential applications of multiphoton absorbers, in the present work we have studied the symmetry-relaxation effects in one- and two-photon absorption spectra in two bichromophore systems based on difluoroborate core linked by biphenylene or bianthracene moieties. We have employed a palette of experimental methods (synthesis, one- and two-photon spectroscopy, X-ray crystallography) and state-of-the-art computational methods to shed light on how symmetry relaxation, a result of twisting of building blocks, affects one- and two-photon absorption of the two studied fluorescent dyes. Electronic-structure calculations revealed that the planarity of central biphenyl moiety, as well as deviations from planarity up to 30-40 deg., ensure maximum values of two-photon transition strengths. Perpendicular arrangement of phenylene units in biphenylene moiety leads to 20% drop in the two-photon transition strengths. More detailed studies demonstrated that equilibrium structures of both compounds in chloroform solution show very different values of two-photon absorption cross sections at absorption band maxima, i.e. 224 GM for and 134 GM for biphenyle and bianthracene linkers, respectively. The latter value is in good agreement with experimental value obtained using Z-scan method. The difference in two-photon absorption cross section between both compounds can be rationalized based on equilibrium geometry differences, i.e. interplanar angle is 35 deg and 91 deg in the case of biphenylene and bianthracene moiety, respectively. It is thus not beneficial to introduce conformationally locked central linker based on bianthracene moiety.

QD:Puf Nanohybrids Are Compatible with Studies in Cells.

Colloidal semiconductor quantum dots (QD), as well as other nanoparticles, are useful in cell studies as fluorescent labels. They may also be used as more active components in various cellular assays, serving as sensors or effectors. However, not all QDs are biocompatible. One of the main problems is their outer coat, which needs to be stable and to sustain hydrophilicity. Here we show that purpose-designed CdSe QDs, covered with a Puf protein, can be efficiently accumulated by HeLa cells. The uptake was measurable after a few hours of incubation with nanoparticles and most of the fluorescence was localised in the internal membrane system of the cell, including the endoplasmic reticulum and the Golgi apparatus. The fluorescence properties of QDs were mostly preserved, although the maximum emission wavelength was slightly shifted, and the fluorescence lifetime was shortened, indicating partial sensitivity of the QDs to the cell microenvironment. QD accumulation resulted in a decrease in cell viability, which was attributed to disturbance of endoplasmic reticulum performance.

Pr3+ doped NaYF4 and LiYF4 nanocrystals combining visible-to-UVC upconversion and NIR-to-NIR-II downconversion luminescence emissions for biomedical applications.

Lanthanide-doped fluoride nanocrystals (NCs) are known to exhibit unique optical properties, such as upconversion and downconversion luminescence (UCL and DCL), which can be employed for various applications. In this work, we demonstrate that by doping praseodymium(III) and ytterbium(III) ions (Pr3+ and Yb3+) into a nanosized fluoride matrix (i.e. NaYF4 and LiYF4), it is possible to combine their UCL and DCL properties that can be concurrently used for biomedical applications. In particular, the emissive modes combined in a single nanoparticle co-doped with Pr3+ and Yb3+ include DCL emission (excited at 980 nm and peaked at 1320 nm), which can be used for near infrared (NIR) DCL bioimaging in the NIR-II window of biological tissue transparency (∼1000-1350 nm) and UCL emission (excited at 447 nm and peaked at 275 nm) that can be employed for germicide action (via irradiation by light in the UVC range). A possibility of the latter was demonstrated by the denaturation of double-stranded DNA (dsDNA) into single-stranded ones that was caused by the UVC UCL emission from the NCs under 447 nm irradiation; it was evidenced by the hyperchromicity observed in the irradiated dsDNA solution and also by a fluorometric analysis of DNA unwinding (FADU) assay. Concurrently, the possibility of NIR-II luminescence bioimaging through biological tissues (bovine tooth and chicken flesh) was demonstrated. The proposed concept paves a way for NIR-II imaging guided antimicrobial phototherapy using lanthanide-doped fluoride nanocrystals.

Polymeric Nanocarriers with Luminescent Colloidal Nanoplatelets as Hydrophilic and Non-Toxic Two-Photon Bioimaging Agents.

INTRODUCTION: Semiconductor nanoplatelets (NPLs) are promising materials for nonlinear optical microscopy since they feature good two-photon absorption (TPA) properties, narrow photoluminescence spectra and high quantum yields of luminescence. Nevertheless, the use of semiconductor NPLs is inevitably connected with concerns about heavy metal ion toxicity and their intrinsically hydrophobic character. METHODS: Our contribution focuses on the design and engineering of coloidal bionanomaterial consisting of two-dimensional highly luminescent CdSe semiconductor NPLs loaded into spherical and homogeneous polymeric nanocarriers (NCs) based on poly(ethylene oxide) and poly(propylene oxide) block co-polymer. The biocompatibility and usefulness of the NPLs-loaded polymeric NCs in two-photon induced bioimaging was demonstrated in vitroby cytotoxicity and two-photon microscopic studies using eukaryotic (normal fibroblasts and cancer ovarian) cells. RESULTS: The encapsulated NPLs maintain their intensive and spectrally narrow photoluminescence, as well as preserve good TPA properties, while the surrounding polymer shell imparts hydrophilic character and non-toxicity towards eukaryotic cells. Specifically, TPA cross-sections of the colloidal NCs loaded with NPLs show large values reaching up to 2.0 × 108 GM, with simultaneously two-photon brightness reaching 2.2 × 107 GM at 870 nm. MTT proliferation assay performed on cell lines treated with encapsulated NPLs revealed at least 70% viability of normal human gingival fibroblast (HGF) and cancer ovarian (MDAH-2774) cells, while the results of multiphoton imaging of murine (L-929) fibroblasts suggest that the encapsulated NPLs are capable of labelling the target cells enabling their visualization. CONCLUSION: As a result, we obtained water dispersible and temporally stable hydrophilic NPLs-loaded NCs that offer excellent, both one- and two-photon excited fluorescence preserving optical properties of the raw hydrophobic and colloidal NPLs. The biological responses upon eukaryotic cells indicate that the encapsulation process protects cells from the toxic influence of cadmium simultaneously preserving the unique multiphoton properties of the active cargo which opens a promising perspective for its application in multiphoton cancer bioimaging excited at the "optical transmission window" of biological tissues in near-infrared range.

Multimodal polymer encapsulated CdSe/Fe3O4 nanoplatform with improved biocompatibility for two-photon and temperature stimulated bioapplications.

Multimodal polymer encapsulated CdSe/Fe3O4 nanoplatforms with dual optical and magnetic properties have been fabricated. We demonstrate that CdSe/Fe3O4 nanocapsules (NCs) upon excitation with UV radiation or NIR fs-laser excitation exhibit intense one- or two-photon emission at 535 nm, whereas the combination of an alternating magnetic field and 808 nm IR laser excitation results in heat generation. Since anticancer therapies require relatively high doses of Fe3O4 nanoparticles (NPs) to induce biologically relevant temperature jumps, the therapeutic effects of 0.1 and 1 mg/mL Fe3O4 NCs and CdSe/Fe3O4 NCs were investigated using breast cancer cell lines, ER-positive MCF-7, and triple-negative MDA-MB-231 cells. Improved biocompatibility of CdSe/Fe3O4 NCs compared to Fe3O4 NCs was revealed at higher NCs concentration suggesting safe potential medical applications of CdSe/Fe3O4 NCs. In contrast, 1 mg/mL Fe3O4 NCs were found to be more cytotoxic to MDA-MB-231 than MCF-7 cells through iron-induced oxidative stress, lipid peroxidation, and concomitant ferroptotic cell death. We believe that Fe3O4 NCs-mediated cellular response may be heterogeneous that reflects, at least in part, cancer cell genotype, molecular phenotype, and pathological classification.

Nonlinear Optical Pigments. Two-Photon Absorption in Crosslinked Conjugated Polymers and Prospects for Remote Nonlinear Optical Thermometry.

Nonlinear optical (NLO) pigments are compounds insoluble in solvents that exhibit phenomena related to nonlinear optical susceptibilities (χ(n) where n = 2,3,...), e.g., two-photon absorption (2PA) which is related to the imaginary part of χ(3). Determination of spectrally-resolved 2PA properties for NLO pigments of macromolecular nature, such as coordination polymers or crosslinked polymers, has long been a challenging issue due to their particulate form, precluding characterizations with standard techniques such as Z-scan. In this contribution, we investigate thus far unknown spectrally-resolved 2PA properties of a new subclass of NLO pigments-crosslinked conjugated polymers. The studied compounds are built up from electron-donating (triphenylamine) and electron-withdrawing (2,2'-bipyridine) structural fragments joined by vinylene (Pol1) or vinyl(4-ethynylphenyl) (Pol2) aromatic bridges. 2PA properties of these polymers have been characterized in broad spectral range by specially modified two-photon excited fluorescence (TPEF) techniques: solid state TPEF (SSTPEF) and internal standard TPEF (ISTPEF). The impact of self-aggregation of aromatic backbones on the 2PA properties of the polymers has been evaluated through extended comparisons of NLO parameters, i.e., 2PA cross sections (σ2) and molar-mass normalized 2PA merit factors (σ2/M) with those of small-molecular model compounds: Mod1 and Mod2. By doing this, we found that the 2PA response of Pol1 and Pol2 is improved 2-3 times versus respective model compounds in the solid state form. Further comparisons with 2PA results collected for diluted solutions of Mod1 and Mod2 supports the notion that self-aggregated structure contributes to the observed enhancement of 2PA response. On the other hand, it is clear that Pol1 and Pol2 suffer from aggregation-caused quenching phenomenon, well reflected in time-resolved fluorescence properties as well as in relatively low values of quantum yield of fluorescence. Accordingly, despite improved intrinsic 2PA response, the effective intensity of two-photon excited emission for Pol1 and Pol2 is slightly lower relative to Mod1 and Mod2. Finally, we explore temperature-resolved luminescence properties under one- (377 nm), two- (820 nm), and three-photon excitation (1020 nm) conditions of postsynthetically Eu3+-functionalized material, Pol1-Eu, and discuss its suitability for temperature sensing applications.

The Two-Photon Absorption Cross-Section Studies of CsPbX3 (X = I, Br, Cl) Nanocrystals.

The CsPbX3 nanocrystals (NCs) with X = I, Br, Cl, or the mixture of Br:I and Br:Cl in a 1:1 ratio were synthesized and characterized by TEM, DLS, and XRD. Recrystallization of the small luminescent NCs in the metastable cubic phase into bigger orthorhombic nanocrystals was monitored by XRD and identified as the main cause of the nanocolloid coagulation. The recrystallization also leads to a decrease in the photoluminescence quantum yield (QY) of the colloidal solution and shortening of the emission lifetime. The two-photon absorption cross-section σ2 values calculated from femtosecond Z-scan measurements were compared with those obtained based on the two-photon excited emission technique. These two techniques were shown to be equivalent with the cross-section values calculated per molar mass of CsPbX3 perovskite being in the range of 10-200 GM depending on the halide anions X-. The σ2 values recalculated for the mole of the NCs were in the range of 104-105 GM, which is in good agreement with values previously reported elsewhere and the σ2/M parameter was in the range of 0.01 to 0.33. This study shows the perovskite NCs to be a good nonlinear material with the third-order nonlinear optical susceptibility χ(3) of the NCs on the order of 10-11 esu.

Functional CdS-Au Nanocomposite for Efficient Photocatalytic, Photosensitizing, and Two-Photon Applications.

We demonstrate a low-temperature synthesis of hydrophilic, penicillamine-stabilized hybrid CdS-Au nanoparticles (NPs) utilizing different Au concentrations. The obtained hybrid nanomaterials exhibit photoluminescence quenching and emission lifetime reduction in comparison with their raw semiconductor CdS NPs counterparts. An increase of concentration of Au present at the surface of CdS leads to lower photoluminescence intensity and faster emission decays, suggesting more efficient charge separation when larger Au domains are present. For photocatalysis studies, we performed methylene blue (MB) absorption measurements under irradiation in the presence of CdS-Au NPs. After 1 h of light exposure, we observed the absorbance decrease to about 35% and 10% of the initial value for the CdS-5Au and CdS-7.5Au (the hybrid NPs obtained in a presence of 5.0 and 7.5 mM Au), respectively, which indicates MB reduction caused by electrons effectively separated from holes on metal surface. In further similar photocatalysis experiments, we measured bovine serum albumin (BSA) integrated photoluminescence intensity quenching in the presence of CdS-Au NPs, with a 50% decrease being obtained for CdS-2.5Au NPs and CdS-5Au NPs, with a faster response rate detected for the system prepared with a higher Au concentration. The results suggest hole-driven reactive oxygen species (ROS) production, causing BSA degeneration. Finally, we performed two-photon excited emission (TPEE) measurements for CdS-5Au NPs, obtaining their two-photon absorption (TPA) cross-section values up to 15.8 × 103 GM (Goeppert-Mayer units). We conclude that the obtained water-soluble CdS-Au NPs exhibit potential triple functionalities as photocatalysts for reduction and oxidation reactions as well as materials for two-photon absorption applications, so that they may be considered as future theranostics.

Controlling Two-Photon Action Cross Section by Changing a Single Heteroatom Position in Fluorescent Dyes.

The optimization of nonlinear optical properties for "real-life" applications remains a key challenge for both experimental and theoretical approaches. In particular, for two-photon processes, maximizing the two-photon action cross section (TPACS), the figure of merit for two-photon bioimaging spectroscopy, requires simultaneously controlling all its components. In the present Letter, a series of difluoroborates presenting various heterocyclic rings as an electron acceptor have been synthesized and their absorption, fluorescence, photoisomerization, and two-photon absorption features have been analyzed using both experimental and theoretical approaches. Our results demonstrate that the TPACS values can be fine-tuned by changing the position of a single heteroatom, which alters the fluorescence quantum yields without changing the intrinsic two-photon absorption cross section. This approach offers a new strategy for optimizing TPACS.

Color-coded multilayer photopatterned microstructures using lanthanide (III) ion co-doped NaYF4 nanoparticles with upconversion luminescence for possible applications in security.

We report a method for fabricating predefined photopatterns of upconversion nanophosphors using a chemical amplification reaction for direct writing of films with multilayer color-coded patterning for security applications. To photopattern the nanocrystal film we have synthesized rare-earth ion (Er(3+)/Yb(3+) or Tm(3+)/Yb(3+)) co-doped sodium yttrium fluoride (alpha-NaYF(4)) nanophosphors and functionalized the nanocrystal surfaces by incorporation of a photopatternable ligand such as t-butoxycarbonyl (t-BOC). The surface modification allows photopatterning of the nanophosphor solid state film. Furthermore, upconversion nanophosphors show a nearly quadratic dependence of the upconversion photoluminescence (PL) intensity on the excitation light power, and tailoring of the PL wavelength is possible by changing the lanthanide ions. We have demonstrated the capability of anchoring nanophosphors at desirable locations by a photolithography technique. The photopatterned films exhibit fixed nanophosphor structures clearly identifiable by strong upconversion photoluminescence under IR illumination which is useful for a number of applications in security.

Three-Photon Absorption of Coordination Polymer Transforms UV-to-VIS Thermometry into NIR-to-VIS Thermometry.

Lanthanide-based metal-organic frameworks (MOFs) and coordination polymers (CPs) attract much attention as candidates for optical ratiometric thermometry applications. Thus far, excitation of these materials was mainly performed in the ultraviolet that drastically limits their applicability as sensors, e.g., in tissue biological thermometry. As a remedy for this constraint, for the first time, we leverage a nonlinear optical process, the three-photon absorption property of Eu,Tb-CPs to shift the excitation wavelength from ultraviolet into near-infrared region. Experiments demonstrate that three-photon induced thermometric responses of Eu,Tb-CPs follow excellent optical characteristics similar to those determined for one-photon excitation, yet are not identical. The relative sensitivity reaches a very high value of 2.91%K-1 in the physiological temperature region. We put forward a notion that utilizing multiphoton absorption is a general strategy for realizing NIR-to-VIS remote temperature sensing in practically any CP that is designed for UV-to-VIS thermometry.

Exciton Binding Energy of Two-Dimensional Highly Luminescent Colloidal Nanostructures Determined from Combined Optical and Photoacoustic Spectroscopies.

Reduced dimensionality of structures such as 0D quantum dots, 1D nanorods, and 2D nanoplatelets is predicted to favor the creation of tightly bound excitons stable at room temperature, making experimental determination of the exciton binding energy ( R x) crucial for evaluating the performance of semiconductor nanoparticles. We propose a fully optical approach for R x determination based on a complementary combination of photoacoustic and transmission spectra, using 5.5, 4.5, and 3.5 ML CdSe nanoplatelets as a benchmark system. The absence of excitonic features in photoacoustic spectra allows for probing the band-to-band transition, leading to the band gap determination. Such an unusual effect is explained by efficient re-emission of the absorbed radiation typical for high quantum yield structures, keeping the crystal lattice from excess phonon generation. The determined exciton binding energy for CdSe nanoplatelets ranges from 130 to 230 meV, confirming the presence of robust excitons in highly confined 2D systems.