Sensing and bio-Sensng Research 3 (2015 46-52 Contents liste available at SciencoDieoct ee Sensing and Bio-Sensing Research ELSEVIER journal homepage: www.elsevier.com/locate/sbsr Highly fluorescent CdTe quantum dots with reduced cytotoxicity-A Robust biomarker Jandi Kim®, Bui The Huy®, Kavitha Sakthivel®, Hye Jung Choi”, Woo Hong Joo”, Seung Kyun Shin‘, Min Jae Lee‘, Yong-tll Lee** *Depomet of Chenin. Changwon Nana Unive, Chagwon 4-773, Repub of ore depart of Bay, Chow Nationa! Univers, Changwon 64-77, Republi of Rerea "Depart of Agphed Cherry, ol of Ape Sree Ryne Hes Unies, Yong 449-707, Republi Korea ARTICLE INFO ABSTRACT Koon: ‘Ceystene (Cy) capped Cae quantum dats (CaTeaCys QDs) were successfully synthesized in an aqueous outcne ‘medium. The synthesized CATetCys samples were analyzed using Fourier transform infrared (FT-IR) cate spectroscopy, fluorescence (FL spectroscopy, transmission electron microscopy (TEND, confocal micros ‘Quantum éos Copy and subsequently subjected fo the antibacterial test. Systematic investigations were catied ou for ‘otarence reactivation the determination of optimal conditions namely the ratios of CA'e, Cae: Cys, pH value ad he chemi MIT assay "Stablity of CaTe@Cys. Moreover, the reactivation of. ntensity in the CATed@Cys sample was done easly by the addendum of Cys. The introduction of addtional cysteine tothe CdTe@Cys QDs sample showed an ‘ehancement in ters ofthe FL incensty and stability along with the reduced antibacterial activity This ‘was futher confirmed through Thisaoyl blue teteazolium bromide (MTT) assay. Both the result ofthe bio stably tests namely the antibacterial test and MTT assay displayed silaiies betwen che exter pally added Cys and cytotoxicity ofthe bacteria and human HeLa cancer cel lines. Confocal microscopic Images were captured for che CaTe@Cys conjugated Escherichia col © 2014 The Acthors. Published by Elsevier BV. Thisis an open access artcle under the CCBY-NC-ND Hcense (itp jereaivecommons orgliensespby-nend/4 0) 1L Introduction Cenerally, quantum dots (QDs) are referred to as the zero: dimensional colloidal crystals that possesses strong size depen dence and multi-colored luminescence properties [1.2] along with its intrinsic features, such as the sharp and symmetric emission, photostability and high quantum yields. Owing to these inherent inimitable properties, QDs play a vital role in various avenues namely the identification of the chemical moieties [3], clinical diagnostics [4-5], optoelectronics | 10.11), bio-imaging and bio sensing [12-19], QDs are chemically unstable and have poor solubility in water and due to these deficits it was not preferred for biomedical appli- ‘ations, But i associates the promising features needed for the bio- medical studies and hence, a number of attempts were put forth, to make it water-soluble. Primary works relied on the usage of differ- cent thiol stabilizers to promote the water solubility and chemical stability in the water phase. The thiol linkage serves as the protec- tion layer to the core and thus prevents it from surface oxidation ‘Thus, the solubility of the QDs in water can be enhanced. Thiols are often sorted as the ideal choice for the modification of QDs {due to its facile functionality [20]-A wide variety of quantum dots are available namely cadmium selenide (CdSe), cadmium sulphide (CdS), zine sulfide (2n8), etc, [21,22]. The syntheses of these mate als require high temperature, non-aqueous medium and possess low efficiency, These discrepancies can be rectified using cadmium telluride (CaTe) QDs as they are expected to show a high fluores cence efficiency and goad stability [2]. In order to obtain an opti- ‘mum quantum efficiency the following stabilizers are used fatnely meteaptoacetic acid (MAA), glutathione (GSH) and thio- lyeerol (TGA) [23.24]. Few of the works reported the toxicity of CalTe QDs in relation to the metal ions and stabilizers used for is synthesis [25-31], Hence the toxicity level of the stabilizers has tobe considered asa very important factor for biomedical applica tion, where the best choice is confined to the low toxicity. Recent studies relied on the preparation of CdTe using cysteine asa surfac- {ant [52,33] -Cysteine (Cys) isa simple amino acid that possesses the thiol(-SH) group and two other active functionalities (NH, COOH) which is considered to be the essential component of human metabolism and enzyme reactions. Specifically, the pres-4. Kio Sensing ond Bo SetingRewarch3 (2018) 40-52 the complex structure of proteins and is subjected to oxidation during the alteration of the disulfide bond [34], Yang et al attempted the direct labeling of Escherichia col(E. col) using CaTe ‘QDs by the following treatments namely chloroform-SDS, Iyso- zyme-EDTA and osmotic shock and, the results obtained showed an enhancement in the sensitivity [35] ur present work is based on the synthesis of ys capped CaTe ‘QDs in an aqueous medium for labeling the cell and the reactiva tion of FL intensity using additional Cys. Li et al. also reported the recovery of FL intensity of Cae to its inital level by the add tion of Cys [56]. But our research differs from the other reported works in many aspects. Herein, we carried out a systematic inves- tigation on the influence ofthe additional Cys to CdTe@Cys in the facet of biomolecular protection. The results obtained proved the optical superiority of the adopted technique. Addition of Cys to higher concentration significantly improved both the stability of bacterial communities and FL intensity of CdTe@Cys QDs. Further- ‘more, the reactivation ofthe CaTe@Cys QDs resulted in a mitigated activity of CaTe inthe antibacterial test. Along with the lower cyto- toxicity, reactivated CdTe@Cys QDs labeled E.coli were found to be more adequate. MTT assay was earied ou for the determination of the cytotoxicity of reactivated CdTe@Cys to HeLa cells. The results derived out ofthe study proved the complementary evidences for its strong implication as a novel labeling method. Thus it will have 2 promising future and give an unprecedented insight by the way of an efficient labeling of cell using QDs, 2. Experimental 2.1. Reagents and materials Tellurium powder, sodium borohydride (NaBH,). cadmium chloride (CdCl), Cysteine, ethylenediaminetetraacetic acid (EDTA), phosphate buffered ‘saline (PHS) were obtained from Sigma-Aldrich (USA) of analytical grade. Lysozyme from chicken egg white of molecular biology grade was obtained from Sigma~ Aldrich Chemical. Sucrose of ACS reagent grade for microbiology ‘was obtained from Fluka. Deionized distilled (D1L) water was obtained from 2 Milli-O water purification system. All other chem- icals and solvents used for the QDs synthesis were purchased from Deahan Co, (Korea) and are used as such without further purifica~ tion, Luria-Bertani agar plates were obtained from Difco (USA), Filter paper discs were purchased from Toyo Roshi Kaisha, Ltd apan). MTT assay kit was purchased from Biosesang (Korea) 22. Preparation of CaTe@Cys QDs ‘The synthesis of CdTe QDs was carried out in accordance to an earlier reported procedure [27,38] and it consisted of two steps. In the former step, 30 mg of CdCl, was dissolved in 100 mL of D1 water in a three necked flask, and to it added 363 mg of Cys care fully under ultrasonication. The resultant solution was adjusted for the desired pH using 1M NaOH solution under vigorous stirring On the other hand, Te precursor solution was prepared using Te powder and NaBH, Typically, 20 mg of Te powder was mixed with 30 mg of NaBH ina vial bottle with 0.6 mL DL water. Then the vial bottle was sealed with a rubber stopper and heated at 50°C for 30 min. This process resulted in the generation of, NabYTe activated solution and it was withdrawn using a syringe. tn the latter step, Ca precursor solution was heated to 100°C approximately; at this point the prepared Te! precursor solution was injected into the Ca" precursor solution, The mixture was subjected to reflux at diferent reaction time interval in order to control the optical properties of the CdTe QDs. After completion of reaction the mix- ‘ure was brought down to ambient temperature. Then the precip- ftation of the CéTe QDs were done using iso-propanol and subjected to centrifugation at 4000 rpm. The purification process ‘was repeated several times using iso-propanol[water. Finally, the product obtained was dried overnight at 80°C and thus yielding the desired product namely, CaTe QDs, 23, Reactivation of C&Te@Cys QDs Recovery of fluorescence intensity to its initial level using Cys, ‘was previously reported by Li etal. [36]. Our experimental wrk ‘was built on this platform with slight modifications. Samples of the as-prepared CdTeaCys and Cys were prepared at different ‘molar ratios viz 1:1,1:2, 1:3, and 1-4. Then, the mixtures were sub- jected 10 sonication at raom temperature for duration of min. The FL intensity of reactivation states of CTe QDs was analyzed, 24, Characterization techniques Fluorescence spectra of QDs were recorded using a spectroflue- rometer (FP-6500, Jasco, Japan) at an excitation wavelength of A B o. o © xo. i 2 20. : : 2 0. YK °. 0“ |. im ea Secing and Bo-SosingResvrch 32015} 46-52 A B wo + 0m; 205 sme 200 ene “ -> NO PHS Te. — n 2 * i ie be a z \ WW o™. \ i ° Zena ae ero vn ig. 2.) Chemical stabity of CTe@cys QDs prepare 2 various p(B) uence of akiona ys on the FL intensity of C#Te@Cys Qs. (The molar ai of CHTe@Cys (Odeacetonsl evs was 1) a): anginal CATe@tyx QDs wthoeadtonl Cys molar ati of adeitionl Cy ere (B16) 2583 (e4,respectve The inset ves Image ofthe adding Cys soltions unger UV amp (before ane ser 355 nm. Fourier transform infrared (FT-IR) spectra were obtained using a FIR spectrophotometer (FI/IR-6300, Jasco, Japan), Trans- mission electron micrographs were captured using afield emission {ransmission electron microscope (IEM-2100F, JEOL, Japan) at 200 KV, Fluorescence microscopic images were taken using a Laser scanning confocal microscope (FV101, Olympus, Japan), Relative absorptivity ofthe cells were observed using UV-vis spectroscopy (Bio Tek, USA) 25. Evaluation of antibacterial activity Antibacterial activities of the synthesized compounds were determined using disc diffusion methods [39], Primarily, a 100 ul. suspension of E.coli IMSNU 10080 (ATCC 10798) (1.5 x 108 CFU im) was subjected to inoculation in a Luria-Bertani (LB) agar plates. Then the filter paper discs (6 mm) were placed over the inoculated plate, and to it added 15 ul of the compounds, and incubated at 37 °C for 24h. 26, Bacterial treatment and QDs labeling E coli cells were washed twice with phosphate buffer solution (pH 8) and was treated using Lysozyme-EDTA [3540], The “Hl prepared cells were resuspended in buffer A (20% sucrose, 0.03 M ‘Tris-HCl, EDTA 10 mM, pH8). After 10 min of incubation at room, temperature, cells were resuspended by lysozyme (the sample of a final concentration were 2mgimt) followed by incubation at 37°C for 20 min. Then the sample was centrifuged at 10,000 rpm for min and washed twice with buffer B (S0mM Tris-HCI, (05M sucrose, pH 8), Finally. E.coli cells were resuspended in buf- fer €(50 mM Tris-HCI, pH 8) and then mixed thoroughly with the prepared QDs. After shaking at room temperature for 4h, the cells were washed twice with bufler C 22.In vitro cell viability Cll viability was checked by the standard MTT assay with slight modifications to the earlier reported procedure (41 Briefly, the cells were grown in 96 wel plates at 37°C in a humidified atmosphere of 95% air/5% CO» After the treatment of the extra Cys at various concentrations, thiazolyl blue tetrazolium bromide was added to each well containing the eels (inal cone. 05 mg! ‘ml) and incubated for 2.5 h. Dimethyl sulfoxide was added to sol- ubilize the blue MTT-formazan product and the sample was incu- bated for further 30 min at room temperature, Absorbance of the solution was recorded at a wavelength of 550 nm, ig. 4. Sehenate representation depicting the eactvaton steps of CATE@CYs on adton of Cs. (A) Bae Ca" fons an (8) Bae Ci” tansadoal CVs showing the sttengthened FL intensity.4 Bie Seng and Bo Sesing Recut 32005) 46-52 43, Results and discussior 3.1, Characterization of CdTe@cys 3.11, Structural properties Fig. IA depicts the FAR spectra of free Cys (a) and CaTe@cys ‘QDs (b). The peaks observed for the CdTe@*Cys QD sample at an intensity of 1549em~* (asymmetric stretch) and 1304em"' (symmetric stretch) corresponds to the presence of COO™ group. ‘Another band located at 2900cm™ corresponds to the C-H stretching vibration (asymmetric, symmetric) of Cys in CdTe@cys, QDs. The band located at approximately 3286¢m' and 3330cm"" can be assigned to the N-H stretching. Also, the peak around 1598 cm" can be correlated to the N-H bend since Cys hhas an amine group [3642|. The absence of the peak at 2550em ? confirmed the formation of Cys capped-CalTe QDs. It indicates S-H stretches (symmetric) and thus confirming the formation of covalent bonds between thiols group and Cé?" ions ‘on the surface of CaTe QDs, 3.12. Influence of molar radio und acidity on the optical properties ‘The Emission spectra were recorded for determining the opti- mized molar ratios of Cd:Te:Cys and the pH value. Fig. 51 depicts the emission spectra of CdTe@Cys QDs sample which show a strong dependence of FL intensity against the change in Te" molar precursor solution while the concentations of Cys and Ca” were Fixed to be 4 and 1, respectively. ‘The higher degree of QDs luminescence can be correlated to the presence of excessive cadmium monomers in the initial stage of nanocrystals growth (3). The intensity of PL showed 2 steep increase with increasing concentration of Te* and the further raise in Te concentration leads to a decrease in intensity. When the molar concentration of Te?” was 0:24 the FL intensity reached maxima. The presence of excess Te* paved the way for oxidation ‘and in-turn attached itself to the surface of QDs. Ths resulted in ‘the appearance of defects on the QDs surface which in-turn dimin- ished the FL intensity of CéTe, ‘The surfactant plays a pivotal role in the synthesis of CdTe QDs, and henceforth the molar concentration ofthe surfactant was op mized by maintaining the ratio of precursors (Cd:Te) as constant. ‘The maximum FL intensity of CdTe was observed, when the molar ratio of Cd:Te:Cys was maintained to be 1:0.24:4, ata fixed Cé: Te (1:0.24),as shown in Fig. 1B. A further increase in Cys concentra- tions to 6 and 8 give rise to the redshift in the emission peak. This may be attributed to the presence of excessive Cys molecules, which impede the formation of CaTe QDs and finally afix itself Fig 4. Ansbactcal acct of addtional cs introduced aTeseys QDs on E col (ls {arg 000,016, 18, 18, 54,50, l4¢ngln of aon Cy ape contol oun) to the Cys andor Ca?"on the CaTe QD surface. A recent report by Omoho's group portrays the QDs of Te’'-rich surface, where a lower quantum yield was observed than that of the C@-rich sur- face. This may be ascribed to the density of uncoordinated Te fons that gives rise to the process of hole-trapping | 44, ‘Another important parameter to be considered in the aqueous ‘media synthesis of quantum dotsis the pH value ast controls both the emission wavelength and PL quantum yield Three different pH values namely pH =4, 8 and 11 were chosen for the synthesis of CdTe@Cys QDs and their PL spectra, shape and size were studied in detail. The pH value of precursor solution of as-prepared QDs samples was controlled by 1N NaOH solution. This experiment ave a strong evidence for the influence of pilin determination of the maximum emission wavelength of CéTe@Cys QDs. A com- plete depletion in the PL emission intensity was observed at a pH value as lower as 4 or at a higher pH value of 11. But, at the pH value of 8, emission intensity was considerably strong (results not shown}, Transmission electron micrograph (TEM) reveals the complete profile of the particle size and the state of aggregation of Cys capped Cae QDs, and is shown in Fig S2. There are translu- cent gray areas for the surfactant of organic Cys on the edge of particles. The average particle size of CdTe@Cys QDs was about ‘Snim with a good particle size distribution, 3.13, Fuorescence stability of CdTe@Cys QDs In order to extend the application of CéTe@Cys QDs to bio- labeling studies, a systematic investigation of FL. intensity at vari- (ous times for the CdTe@Cys QDs prepared at various pH |45] was cartied out in a phosphate butler saline as medium (Vis. 2A). ‘The intensity of PLwas decreased to a formal are shape for pris- tine CéTe@Cys QDs in the absence of PBS. Ata pH value of 6, the FL. intensity of PBS that contains CaTe@Cys QDs was almost quench ing, and a gradual aggregation of nanoparticles was visualize. Few of the researchers have reported, the absence of aggregation ata pH value of 8 [45), The brightest FL. intensity was found within this range. The quenching patterns of CaTe@Cys QDs in PBS (pH 7) ‘were in analogue to that of pristine QDs; however a weaker FL intensity was noted, Ata pH value of 8 the decrease in FL intensity ‘was noted after an hours' duration. But after an interval of 2, an enhancement in the intensity was observed when, compared to that of the original intensity of the unmingled QDs. After 24h, the FL intensity of CdTe@Cys in PBS at pH 6-7 and pristine CaTe@cys were almost quenched, However at a pH value of 8, Fl intensity of CéTe@Cys in PBS was found to be higher than that of pristine CaTe@Cys, Therefore, further experiments were preceded Using PBS dispersed CdTe@Cys prepared at a pHl value of 8. 3.2 Reactivation of fluorescence in CATe@Cys QDs 3.2.1, Influence of additional Cys on FL intensity of CaTe@®Cys QDs ‘The surfactants play a significant role in protection of the sur~ face and core from oxidation and subsequent degeneration. Few of the notable surfactants are TGA and mevcaptopropionic acid [20]. Recent reports revealed the possible recovery of FL intensity fof CaTe QDs to its initial level by the addition of Cys (36). There- fore, we investigated the efficacy of supplemental Cys on CdTe@Cys by the addition of Cys solutions at different molar ratios and the FL intensity was recorded. However, we inferred that the addition of Cys to the CdTe@Cys QDs resulted in a steep increase ff FL intensity when compared to that of the as-synthesized CaTe@Cys QDs (Fig. 28 and Fig. 3). ‘Additionally, we observed the change in color forthe additional Cys introduced CaTe@Cys QDs sample even with the naked eye ‘under visible light. A further increase in the amount of Cys led to the red shift which is notable by the peak value of the emission spectra of additional Cys-introduced CdTe@Cys QDs. The FLso 1. im ea Sensing an -Sesng Resvrch 3 (2015) 4-82 intensity of ad ‘optimization after 5 min when the ratio of CeTe@Cys:Cys was fixed tobe 1:1 nal Cys-introduced CéTe@Cys QDs reached 3.22. Photoluminescence stability of additonal Cys introduced CaTe@cys QDs Systematic photoluminescence stability was studied for the additional Cys introduced CaTe@Cys samples. Typically, five sam- ples were prepared with various molar ratios of CdTe@Cys QDs to supplemental Cys, namely 1:1, 1:2, 1:3, and 1:4 along with ‘the pristine sample as reference. The stability of pristine and addi tonal Cys introduced CaTe@Cys QDs were evaluated by analyzing its FL intensity at various time intervals from 5s to a week at pH & in PBS medium. It can be clearly observed that FL intensity of CaTe@cys QDs rose steeply with increasing concentration of add tional Cys. But in the case of pristine CaTe@Cys QDs sample the FL Intensity was substantially quenched after 1h. Fig S3 depicts the FL intensity of additional Cys introduced CaTe@Cys samples in which it showed an instant recovery and thus contributes to the increased! intensity after a time period of 2h, But, it dwindled after a days’ time. In the case of samples with the ratios of 1:3 and 1:4, the FL intensity was observed to be stronger even after a week, 3.3. Application 3.3.1. Bffects ofthe additional Cys to CaTe@Cys QDs on antibacterial ‘activity and in vitro cell viability The antibacterial effects were evaluated by the addition of Cys and the role of excessive Cys was predicted in the case of Esche- richia coli (E coli) cells, Fig. 4, depicts the efficiency of the CaTe@cys QDs towards the execution of bacteria. However there is a decrement in the antibacterial activity with an incremental concentration of additional Cys (sample a-h in Fig 4). Cytotoxic level of E.coli was completely eliminated at a Cys concentration of 18 ngimL. Hence, the survival of . coli follows a concentra tion-dependent inclination towatds the added Cys [47]. Therefore, itcan be well established that, the Cys addendum isthe key factor inthe reduction of antibacterial capacity; and the concentration of added Cys should reach a particular level for an efficient interfer tence in the antibacterial elfcacy of CaTe@ys QDs. Further investigations were carried out with respect tothe color variation of CdTeaCys QDs solution to that ofthe externally intro: duced additional Cys. Primarily, the as-synthesized CdTe@Cys QDs solution was colorless but after the addition of Cys it turned into red. This phenomenon may be ascribed to the formation of a ‘new complex by the introduction of additional Cys to CdTe@cys QDs. Arelatively lower oxidative ability was discernible in the case of Cys reactivated CaTe@Cys QDs and thus cause only a meager amount of damage to the bacteria, when compared to that of the bare CdTe QDs and CdTe@Cys without introduction of additional Cys Also, in vitro efficacy of extra Cys was examined by cell viabil- ty assay using a human cervical cancer HeLa cel line (Fig. S). In this work, we substantiated the enhanced proliferation sta bility in the living cells as a function of additional Cys concentra tions. As the concentration of additional Cys increased, an increase in absorbance was noted. It gives 2 clear picture of the increased levels of living cells than at a Cys concentration of OmgimL When the concentration of Cys was Smgiml, an enhanced stability was observed. The results obtained from both the bio-stability studies namely the antibacterial test and MTT assay are in good agreement with each other in terms ofthe viabil ity of the bacteria and living cells. The Cytotoxicity factor related to the cadmium metal jon has been eliminated. The presence of excessive Cys gives rise to the formation of complex via thiolate coordination between the cysteine residues and cadmium ions ‘The presence of additional Cys are extremely competent in theLie a Sensing ond Bo Sensing Research 3 (2018) 46-52 s process of cadmium fons capture, whereby the ability to block cad~ jum toxicity was increased in vivo [48]. All these facets namely the disappearance of antibacterial eflect and the sharp improve- ‘ment in cell viability reveals the reduction in toxicity. 332, CdTe@Cys QDs labeled E.coli cells Confocal microscopic images also demonstrated an enhance ‘ment in the fluorescence intensity by the addition of Cys to that ‘of the labeled bacterial cells, Initially. the pretreated coli cells ‘were labeled with both the samples namely the pristine CdTe@Cys ‘QDs and additional Cys introduced CaTe@Cys QDs based on an ear lier reported procedure [34]. Lysozyme plays a significant role in the membrane fluidity and permeability of the cell membrane and thus promotes the QDs to enter into cells for labeling [35.40]. When E, coli cells were treated with the pristine CdTe@cys, ‘QDs, an absence of fluorescence was observed inthe bacteria ells. ‘On the other hand, when the E.coli cells were subjected to treat. ‘ment by the additional Cys introduced CaTe@Cys QDs an enhance- rent in the fluorescence was observed and this may be attributed to the introduction of additional Cys molecules (Fs, 5). 4. Conclusions In summary, we have successfully demonstrated a novel method for the reactivation of degenerated FL intensity along with the reduction in antibacterial activity of CdTe@Cys QDs using addi tional Cys supplementation. The influence of precursor’s ratio, pH value and concentrations of additional Cys were studied systemat- ically and optimized. Along with an enhanced chemical stability a strong FL intensity was recorded at a pH value of 8. I was inferred that an additional Cys is required for CéTe@Cys QDs samples to bring about the requisite enhancement in FL. intensity when com- pared to the pristine CdTewCys QDs. Additionally, an impeded ‘ytotoxicity of CdTe@ys QDs was discernible due to the introdue- tion of addtional Cys. Addition of the higher concentration of Cys plays a vital role in the enhancement of the bacterial activity of {CATe@Cys QDs and cell viability. Antibacterial studies, confirmed that the addition of Cys at a concentration greater than 18 ng/mL. Ihave restricted the antibacterial effect. Cell viability studies also ‘confirm the reduction in cytotoxicity at a concentration greater than 5 mgiml of additional Cys. Confocal microscopic images also proved its efficiency. All these results provided significant evi- ‘ences fori to be utilized as an efficient labeling agent forthe tar- sgeted cells, Conflict of interest ‘The authors declared that we have no conflicts of interest to this work ‘Acknowledgments ‘This work was supported by the Regional University Research Program (NRF 2014-08793) and the Priority Research Centers, Program (NRF 2010-0029634), through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology. Appendix A. Supplementary data ‘Supplementary data associated with this article ean be found, in the online version, at hitp|/dxdoi.org/10.1016)jsbst-2014.12.001 References 111M, Aawond, ML. Steigeald Le, Sus, The quantom mechanic of are Semiconductor esters "quanta ds) Anau Rev. Phys: Chem 81 (2990 {2} SE. Waiste,L Swart & van Drie SG. Hickey, ¢ ée Mello Donegs. Highly Temnesen watersinbe ele quantum got Nano Lett? (2005 903-307 Ip) YUL ¥- Zhou, 1% Wangs. Pette, YZiae,Z Tang C. Ne Chilly of uation site coling ales the ettancy of euantun das Angew Gem ine 80 (2011 s850~5854, [a) i Ceseke- Mertz M. Monte, Quantum dos as versal probes in medic Sciences: synthesis Mater Sing 352013) 1006-102. Is) ¥.Baaliot Chang, Levy-Nsseabaum, 5. Jon RW Kall, Langer, 0 Forcihzad, Quantum dot-apraner onjgates er syachonous cancer ng, therapy. ava Sensing of ug livery based on Br-fuorescence reson Jo) Mt zhang. lng co, iLL. Cuan € Sun. Lu, Rapid determination of Telamin In mii uang water-soluble CéTe quan dot a fuorescence Probes food Ade Contr Part Chem Anal Control 29 (2012; 133-348 (7) EMGhamed ¥en Ha J ng enti Po" detection by Ie) ¥hen chen ¥ Hei Un, sheng, CLS Lua, Q Ca Cstinecappee (ate Go-based sensor for snp ap Selaciveeeecuon of nicole, Ip) EP oven Maines, Maina Cara, Re Keatownll, A Ru Medina, ‘multicommmutated flow injecion analysis Talanta 109 (2013) 203-208, Ino) x Geng Nis Xing Sone Gs Sun Gch, Zong. 2 it Z dane Soni Xe i guint another, tproctectronie ns Ad Mater, 222040) 638-642, tru Sieh DEN Wong te Pang engi mnie aeen ‘Sf eopper i one Talsnta 1102012] 314-518 [na] Rs. Couhan. JH, Naz A Quresn.E co-qantum dot biocniuates as ioe el uorescentreparers for probing cell eamape.| Mater Chem B Uh ede |e Sica, X Chen, Nanopore tig [04] & Chang. X Yang. F Wang, ¥. Wang Yang, N. Zhang. 8 Wang. Water soluble Auoresceace quantum de rode being ver cancer cei, Mater Sl Mater (05) LE Medina, AT. Uyeda, ER. Goldman H. Martouss, Quantum dot Dioceses for imaging labeling abd sensing. Nt Mate. 4 (2005) 435 116) Mak So.6.X, AM Loening SS, Gani Rao, Samoan quantum a7) X ichale: Fae Ueto ays Doase, UG Sundacesan A Wu 5 Gamphi.s. Wess, Quantum dos for ve cel vito lnabng, and famnnmes Scene 3072005) 538-548, {ijoresceat manopaticies fr cel labeling J Mater Cheas. 8 1 (2013) 2315- (09) ALL Nu ¥.ULF Yang X Su, A novel pcylodein-ds opal biosensor [20] SA Owens, Mc Carpente LW Sonne CA Mle, Renan, CA Ogoakr, En Meng, D1 hes, Reversee-phase HPLC separation of wate-solibe onset ann as, Phy, Cem 15 201) 1852 f 1 th ti 1 ok ep nde ch vn a0 ns fF YM a. ea ee (25) ¥. Su, Mt Hu, Fan ¥ He 1, ULM. Wang. P Shen @ Muang Te ‘eleased cadmium ions and nanopatiie properties, Biomaterials 31 (2010} ‘eros (tan Fao, Sa. STO Masha Coe oan (an tin eta x Yaa. py fact cau fa un ng’ ua. ac ng Suc gnddepenet cl itn abet atone ery pomerconce ccs dhe se 0 (07se 4 Ki Sensing an Bi-Sensing Research 3 (201) 46-52 [29] 8A. Rezalins, JS. sto. amium-conaining nanoparticles: perspectives ‘an pharmacology an toiclony 9 guantm do Tova Appl Pharmacol 238 (2008 280-28 [30] Ea contreras. ho, zhu, HL Pappas, Gs, W. Zhong. VA olin “oxity of quaniom dots ane cadrun salt to Caeordds legos ater Imulegeneratina expose nv Sl Techno 47 (2012) 1348-1754, (a) C wang Zheng ¥ tang. ML Co, | Huo, Du % Mao, D. Zhou. Rape fetermination ofthe tony of quantum dts with lamin bacena evar Mater 1772010) 1154-1137, [32] USiex Mot D1 oven, EM. Phares, MC Cabeza 8, Quintero Chacaesaton of estes capped Cae quantum dets ad appleaton to fest cl) dfency In boil samples from cca Il patent. Ata {Gm Aca 785 (2013) 11-118, [33] ¥ Wu, Zhang 8. Sun One pot aqueous synthesis of nar inared emiting is quantum dos, ppl. suet Se. 258 (2012) 7181-7187 (34) Z wel USun,.)2 Zhang 1 Yang ¥-Yane L sh Chene nadie eae furth uprcotvertg aabopitides for Us Wire and in Wo Boimaste Siomateals 352044) 387-302 (35) C Yang We. ¥ Ls J Zia, B.S, Dee and apd quant dots [being of Echt Cot els, Cala Interiace S393 (2013) 436-448 (36) MCL zo zhao Sun YM Wang 2 Done 8 Xu, reparation an Dusteauon of Catete capped ae quantum dots ands see secoer of GesenteArcee,|in 130 (3010) 095 aga HHyetotheal sys of til capped Cae nanoparticles ab thei opel properties, Pays Chem. Chem Pls. 13 2015) 2903-2011 [38] 5 Chen] Tan ¥ Jang. Y Zhao, | Zhang. 8. Thao, A one step selective Iioresceace turn-on deteton a tine an homacyaeine based on Tac i an ats pertinence 7872013) 7 199] PS. Morey) Baon. MA Piller, PC Tenover RH Yoke Mansa of iia Miroiioy. th ASML Washington, De. 1995, pp 1527-1538 Mol Ps. Wang XL Sacer on cel tne quant dr syresied in gueous Sotin for Boop! labeling, Pays. cher. C115 (2009) 7670~ wore. IDS tee. Kit, HR Lee, MI. Uae, ¥, Shiba, ¥ Oishi AS. Kim, Fypescanchea ‘double véophllebleck copatye: micelles of pay fethyene oxide) and polyeycerol fr piitesponsive “drag eeivey macromolecules 13 (2012) 1190-1198, [a2] 8 The Huy MH Seo, x Zhang, Y- Le Selecveoposesing of clenbutera panies mpd plmercappe Ae gu (a3) CLL Hla Fang | en, iasian enhancement ofthe quantum ye of Cafe nanocrystals synthesized in aqueous phase by consoling the pH and Concentration of pecuse soln) Latin 116 (2008) 55-68. [ay 8" Omego, JF Aldana CD. Heyes. Radatie and noazadtive Metine fnginceing of goanturn dota mulpe” solvent by_surtace fom Stechomnetsy and lgands, Ps. Chem. 117 (2015) 2517-252. as} Me Cut Bua, Nin © Wey §. Zhang, CZ, taeation of intact Dll cas by “tec Ce tant eis ah (as) an, Lu. Zho.@ XU. Zao, Conbolable sytess and cell imaging {tudes on CT quanto dots togethor appr by satan an tog ie Se] Caled lnetace St 33 2000) 364-508, ar) 2 tac 1, HB, Gan Tok X Yang Mechanise of atineodiat ‘evi of Ce quant dos Lange 2¢ (2008) 5445-5452. [40] jitervand. 0. Leing, Vay Cadman) complex formation with ‘stene and peice tory Chem #82008) 3758-5771.