Self-assembled aggregation-induced emission supramolecular probe for temozolomide detection and cell imaging

Self-assembled aggregation-induced emission supramolecular probe for temozolomide detection and cell imaging | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 2 January 2025 V1 Latest version Share on Self-assembled aggregation-induced emission supramolecular probe for temozolomide detection and cell imaging Authors : Chenrui Jiang , Feifei Chen 0009-0002-8960-9367 , Yue Chen , Yu Sun , Hua He , Pierre Dramou , Tao Xu , and Hongbin Xu 0000-0002-8406-9827 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.173578157.70405911/v1 230 views 166 downloads Contents Abstract Introduction Experimental section Results and discussions Fluorescence mechanism Optimization of detection conditions Analytical performance of fluorescent probe Analysis of TMZ in serum samples Inhibition of cell viability TMZ imaging in cells by CB[7]@TTPE Conclusions Acknowledgments References Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract In the proposed work, an efficient supramolecular fluorescent probe method based on the cucurbit[7]uril@1,1,2,2-tetra(biphenyl-4-yl)ethene (CB[7]@TTPE) was developed for detecting temozolomide (TMZ) at trace levels in serum samples and realizing fluorescence imaging of TMZ. Macrocyclic molecule CB[7] and aggregation-induced emission (AIE) molecule TTPE formed the self-assembled fluorescent probe with high degree of solubility via host-guest recognition. The sensing behavior of nanocomposites toward TMZ was investigated by Fourier transform infrared spectroscopy, nuclear magnetic resonance spectroscopy, ultraviolet-visible absorption spectroscopy, fluorescence spectroscopy and fluorescence lifetime. Under the optimal detection conditions, CB[7]@TTPE nanoprobe can be used for the rapid and sensitive analysis of temozolomide in the linear range of 1 to 20 μg/mL with the detection limit of 0.25 μg/mL. The probe system has excellent selectivity and can be applied to detect TMZ in actual serum samples with satisfactory recovery. Furthermore, CB[7]@TTPE exhibited great intracellular drug fluorescence imaging performance and was successfully used to monitor and report TMZ in human glioma cancer cells U87 and T98G. In summary, the prepared AIE composite displays strong fluorescence and good water solubility, which is suitable as a supramolecular fluorescent probe for the determination of TMZ in actual fluids and also provides a new idea for subsequent biological imaging. Introduction Temozolomide (TMZ) is a second generation imidazotetrazinone derivative with methylation properties [1-3]. As an oral alkylating agent, TMZ is a standard first-line chemotherapy drug for the clinical treatment of malignant glioblastoma such as glioblastoma multiforme and anaplastic astrocytoma. Its therapeutic concentration is about 1-15 μg/mL. The physicochemical properties and small size of temozolomide give it the ability to easily cross the blood-brain barrier [1, 3-5]. Therefore, TMZ has a high oral bioavailability, but its half-life in plasma is very short (1.8 h) with the low protein binding rate (only 15%), and its distribution in the brain is insufficient (17.8%). This requires repeated administration, which affects its therapeutic effect and increases adverse events (headache, seizure, bone marrow suppression, etc.) [6-8]. In conclusion, it is necessary to establish an uncomplicated, quick, sensitive and accurate analytical method for the specific monitoring of temozolomide in biological fluids and living cells to maximize the effectiveness and safety of the drug. At present, the techniques for determination of TMZ in complex matrix include spectrometry [9-11], high performance liquid chromatography [12-15], liquid chromatography-mass spectrometry [16, 17], capillary electrophoresis [6, 18] and electrochemistry [19, 20]. But the above methods still have some limitations, such as complicated sample preparation, expensive instruments, need of professional technicians, long detection time and low sensitivity. The fluorescent probe is widely utilized in photoelectrocatalysis, environmental monitoring, food safety, biological science and other research fields because of its low cost, simple operation, fast response, high sensitivity, real-time detection, and especially nondestructive imaging capability for targeted molecules [21-23]. However, the probe based on traditional organic fluorophores has a small Stokes shift, and exists the aggregation-induced quenching (ACQ) effect and the interference of fluorescence bleaching, which leads to the poor photostability. Different from ACQ chromophores, fluorescent molecules with aggregation-induced emission (AIE) properties can emit significant fluorescence in aggregation and solid states on account of the restriction mechanism of intramolecular motion [24-27]. Nevertheless, the problem of poor biocompatibility caused by unsatisfactory water dispersibility will still restrict the application of aggregation-induced luminogen (AIEgen) in the practical biomedical field. Cucurbit[n]uril (CB[n], n = 5-8, 10) is a highly symmetrical pumpkin-like supramolecular host composed of 2n methylene groups bridged by n glycoluril units. It has two identical electronegative polar carbonyl portals and a neutral rigid hydrophobic cavity, which can selectively combine with guest molecules through various noncovalent interactions such as hydrogen bond, van der Waals force, ion-dipole interaction, hydrophobic interaction to constitute a steady host-guest complex [28-31]. Although there are cucurbituril macrocyclic hosts with different cavity sizes, compared with other homologues in CB[n] family, CB[7] possesses a suitable cavity size to produce excellent binding affinity with the target, and has good water solubility and biocompatibility, so it has been extensively applied in complex biological systems, such as molecular recognition, drug delivery and so on [32-34]. In this study, a novel self-assembled “turn-off” supramolecular fluorescent probe was developed for selective identification and determination of temozolomide by employing the host-guest recognition between AIEgen tetraphenylethene derivative (1,1,2,2-tetra(biphenyl-4-yl)ethene, TTPE) and macrocyclic supramolecule CB[7]. Finally, the proposed probe could also achieve the visual imaging of TMZ in living cells, which has the potential for drug targeted tracking. We have confirmed the AIE characteristics of TTPE and the interaction between CB[7] and TTPE [22]. Experimental section Materials and reagents Temozolomide was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. 1,1,2,2-tetra(biphenyl-4-yl)ethene was obtained from Jilin Zhongke In-depth Research Technology Co., Ltd. Cucurbit[7]uril was acquired from Shanghai Bide Pharmaceutical Technology Co., Ltd. Dulbecco’s modified eagle medium (DMEM) and phosphate buffered saline (PBS) were purchased from Gibco company of America. All materials were used directly without further purification and reagents were analytical reagent grade. Ultrapure water was worked the whole process. Instruments Fluorescence measurement was carried out utilizing a RF-5301PC fluorescence spectrophotometer (Shimadzu, Japan). Ultraviolet-visible (UV-Vis) absorption spectrum data were gained through a UV-1800 UV-vis spectrophotometer (Shimadzu, Japan). Fourier-transform infrared spectroscopy (FT-IR) was recorded by a FT-IR 8400s Fourier-transform infrared spectrophotometer (Shimadzu, Japan). Nuclear magnetic resonance spectroscopy ( 1 H NMR) was generated on a Bruker Avance-500 (500 MHz, Bruker, Switzerland). Fluorescence lifetime was obtained from FLS1000 test instrument (Edinburgh, United Kingdom). The fluorescence images of cells were observed and photographed via NCF950 laser scanning confocal microscopy (LSCM). Fluorescence determination procedure CB[7] (2.33 mg) and TTPE (0.32 mg) were mixed in solid state for 5 min, and afterwards ultrapure water was added to acquire CB[7]@TTPE stock solution. The aggregation-induced emission supramolecular probe CB[7]@TTPE (1.5 mL, 80 μM) based on host-guest interaction was added into phosphate buffer (PB, 20 mM). Then, a variety of TMZ with various concentrations were mixed in the system, and incubated with shaking at room temperature for 10 min. Finally, the fluorescence spectrum of the resultant solution was measured in the range from 400 to 600 nm at an excitation wavelength of 380 nm, and the fluorescence change value F/F 0 was calculated for quantitative analysis (F/F 0 = F CB[7]@TTPE+TMZ /F CB[ 7]@TTPE ). Application of serum samples Briefly, the healthy human serum was diluted 20-fold and a series of temozolomide solutions with different concentrations were prepared with the diluted blank human serum. Then the fluorescence intensity was recorded according to the above-mentioned procedure to gain the TMZ content in each sample. All the tests were repeated three times. Cell viability assay In vitro cytotoxicity of supramolecular system was assessed by the standard CCK-8 method. U87 cells were inoculated in 96-well plates and incubated at 5% carbon dioxide and 37°C for 24 h, followed by treatment with different concentrations of TMZ and CB[7]@TTPE with TMZ for 24 h. After removing the medium, 20 µL of CCK-8 (5 mg/mL) was added to each well and incubated for 1 h. Subsequently, the absorbance value at 450 nm was measured and the cell survival rate was calculated. Cell imaging Human glioma cancer cells U87 and T98G (displayed drug resistance to temozolomide) were cultured in DEME medium with 10% fetal bovine serum in an incubator containing 5% carbon dioxide at 37°C for 24 h. The cultured cells were digested to obtain cell suspension, and U87 or T98G cell suspension with appropriate density was removed and inoculated onto the laser confocal dish. 100 μM CB[7]@TTPE, 80 μM CB[7]@TTPE with a certain concentration of TMZ and commercial dyes were added into the dish respectively, and then the dishes were placed in the incubator for 2 h. Then U87 cells and T98G cells were added with 4% paraformaldehyde and reacted for 15 min, followed through washing with PBS three times. After processing, Confocal microscopy was employed to obtain fluorescence images under blue (405 nm excitation) or green (488 nm excitation) light source. Results and discussions Response of supramolecular probe to TMZ The prepared CB[7]@TTPE probe with a certain concentration has powerful fluorescence intensity. The reaction of temozolomide to supramolecular probe was preliminarily studied by adding its solution into the fluorescence system. It can be seen from Figure 1 that TMZ has the capability to significantly quench the fluorescence intensity of AIE macrocyclic molecular probes, laying a foundation toward subsequent quantitative analysis experiments. Figure 1 Turn off of TMZ for CB[7]@TTPE fluorescence system. Fluorescence mechanism A functional self-assembled supramolecular fluorescent probe was built via the host-guest inclusion between CB[7] and aggregation-induced emission molecule TTPE for monitoring and imaging of temozolomide (Scheme 1). The synthesized CB[7]@TTPE system exhibits great water solubility and appropriate fluorescence intensity. Subsequently, the fluorescence response mechanism between supramolecular probe CB[7]@TTPE and TMZ was investigated by a series of characterization methods, such as FT-IR spectroscopy, 1 H NMR spectroscopy, ultraviolet absorption spectroscopy, fluorescence spectroscopy and fluorescence lifetime. Scheme 1 Schematic diagram of response mechanism between the proposed fluorescent probe CB[7]@TTPE and TMZ. To start with, the FT-IR technique was utilized to explore the host-guest interaction between TMZ, CB[7] and CB[7]@TTPE. The two portals of supramolecular host CB[7] are overspread with polar carbonyl groups, and it can be seen from Figure S1a that its valuable representative absorption peak appears at 1735 cm -1 . The characteristic peaks of N−H in TMZ were 3421 cm -1 and 3388 cm -1 , and the absorption peaks at 1759 cm -1 and 1732 cm -1 corresponded to the C=O stretching vibration peaks of primary amide and tertiary amide, respectively. In addition, the absorption peak observed at 1600 cm -1 belonged to the main special peak of C=C, and the antisymmetric bending vibration absorption peak of methyl C−H emerged at 1452 cm -1 . Compared with the FT-IR spectra of CB[7] and CB[7]@TTPE, the absorption peak of carbonyl group in CB[7]@TTPE+TMZ moved to 1716 cm -1 . It also altered evidently contrasted with TMZ, that is, some typical absorption peaks disappeared in FT-IR spectrum. The above results proved that there was an interaction between temozolomide and CB[7]@TTPE inclusion complex. Secondly, the host-guest recognition between CB[7] and TMZ in aqueous solution was ulteriorly substantiated by 1 H NMR (Figure S1b). Assignments for CB[7], TMZ and its complex are as follows: CB[7] (D 2 O, ppm), 4.13 (d, 14H, J = 15.3 Hz, CH 2 ), 5.44 (s, 14H, CH), 5.66 (d, 14H, J = 15.3 Hz, CH 2 ). TMZ (D 2 O, ppm), 3.89 (s, CH 3 ), 8.53 (d, NH 2 ). CB[7]-TMZ (D 2 O, ppm), 3.67 (s, CH 3 ), 4.08 (14H, CH 2 ), 5.35 (14H, CH), 5.57 (14H, CH 2 ), 8.54 (s, NH 2 ). Compared with the 1 H NMR spectrum in the presence of TMZ alone, the methyl proton signal of temozolomide in CB[7]-TMZ solution system obviously shifted to up-field by 0.22 ppm due to the shielding effect of hydrophobic cavity of macrocyclic molecule CB[7], while the proton response of NH 2 underwent a slight downfield shift. At the same time, the resonance peak of proton hydrogen in CB[7] host also changed owing to the addition of temozolomide. The above characterization consequences nicely present that TMZ can form host-guest interaction with CB[7] and CB[7]@TTPE via hydrogen bonding, hydrophobic interaction and ion-dipole interaction. Then, the fluorescence lifetime of CB[7]@TTPE probe without and with temozolomide was studied. Figure 2 exhibited the fluorescence attenuation curve of CB[7]@TTPE probe. The average fluorescence lifetimes of CB[7]@TTPE and CB[7]@TTPE+TMZ were calculated to be 1.87 ns and 1.76 ns, respectively. The research results displayed that TMZ had no significant effect on the fluorescence lifetime of CB[7]@TTPE composites. Therefore, it can be considered that there is no charge transfer between them. Figure 2 Fluorescence decay curve and fitting line of AIE supramolecular probe CB[7]@TTPE in the absence and presence of TMZ. Finally, by comparing the UV-Vis spectra of TMZ, CB[7]@TTPE and CB[7]@TTPE+TMZ at different concentrations, it was found that TMZ and CB[7]@TTPE only owned the original characteristic absorption peaks without appearing new absorption peaks (Figure 3a). This indicates that there is no ground state complex formed between CB[7]@TTPE probe and the analyte temozolomide, which does not satisfy the prerequisite for static quenching. Hence, the fluorescence quenching mechanism can exclude static quenching. As shown in Figure 3b, the UV-Vis absorption spectrum of TMZ did not overlap with the fluorescence emission spectrum of CB[7]@TTPE, so it can be considered that there is no fluorescence resonance energy transfer effect between them, because of the overlap of donor emission spectrum and acceptor absorption spectrum as a prerequisite. Subsequently, the excitation spectrum of CB[7]@TTPE and the absorption spectrum of TMZ were analyzed (Figure 4). It was observed that the UV-Vis spectrum of temozolomide partially overlapped with the excitation spectrum of AIE supramolecular probe CB[7]@TTPE, which well proved the existence of inner filter effect (IFE) according to the outcomes of spectral overlap. Figure 3 a: UV-Vis absorption spectra of TMZ, CB[7]@TTPE, and CB[7]@TTPE+TMZ; b: UV-Vis absorption spectrum of TMZ and fluorescence emission spectrum of CB[7]@TTPE. Figure 4 UV-Vis absorption spectrum of analyte TMZ and fluorescence excitation spectrum of CB[7]@TTPE probe. Optimization of detection conditions Some influence factors such as pH, temperature and response time of the solution system were researched for the purpose of acquiring the best measurement performance. According to the pH test consequences (Figure S2), the F/F 0 of the solution system was relatively stable and did not vary significantly in the pH range of 4-9, suggesting that it has a wide detectable pH range. Considering that CB[7]@TTPE probe containing TMZ owned the best fluorescence quenching effect at pH = 8, it was finally selected as the subsequent standard detection condition. Next, the incubation temperature was chosen as the variate to explore (Figure S3). The comparative fluorescence intensity of the system was the optimum in the temperature range of 30°C-40°C. When the temperature rose from 40°C to 50°C, F/F 0 increased clearly. This discovery indicates that the growth of incubation temperature is unfavorable to the stability of the probe solution. Based on the above situation, it was decided to conduct the follow-up experiment at 30°C. Moreover, the fluorescence intensity ratio of the system under different reaction times was shown in Figure S4. F/F 0 weakened rapidly within 10 min after the addition of TMZ, and then the fluorescence intensity of the solution tended to be steady. Hence, 10 min was selected as the optimal response time for fluorescence measurement. Analytical performance of fluorescent probe Under the optimum assay procedures, dissimilar concentrations of the TMZ solution were appended to the CB[7]@TTPE inclusion complex. It was observed from Figure S5 that the addition of the target TMZ would affect the fluorescence intensity of the probe system, and the higher concentration of TMZ solution, the more obvious fluorescence quenching phenomenon. There was a good linear relationship between TMZ concentration and relative fluorescence intensity in the range of 1-20 μg/mL. The linear regression equation was F/F 0 = − 0.0099C TMZ + 0.9902 (R 2 = 0.9999) with a low detection limit of 0.25 μg/mL (3σ/S, where σ is the standard deviation of blank samples and S is the slope of calibration curve). Compared with other TMZ analytical techniques (Table 1), the developed supramolecular fluorescence probe method has the advantages of inexpensiveness, simple operation, no troublesome sample pretreatment, high sensitivity, fast response speed and suitable determination range. Table 1 Comparison of other reported approaches for TMZ determination UV spectrophotometric method Centrifugation to settle the lipids 4-18 μg/mL 0.32 μg/mL / Liposomal dispersion [9] Near- infrared fluorescent nanosensors / / Down to 30 μM 6 h U-87 MG human glioblastoma cells and SKH-1E mice [10] RP-HPLC-DAD Liquid-liquid extraction and evaporation using nitrogen 0.2-20 μg/mL 0.084 μg/mL 15 min Rat plasma and brain [13] Capillary electrophoresis Buffer electrolyte pretreatment 10-15 μg/mL 0.33 μg/mL 0.28 μg/mL 1.4 min Water and serum [18] Supramolecular probe Not needed 1-20 μg/mL 0.25 μg/mL 10 min This work The complexity of the actual environment makes it an important index that the supramolecular fluorescent probe can specifically determine TMZ, so the selectivity of this method was investigated. Adding interferents such as amino acids, glutathione, glucose, uric acid, diverse cations and anions into the detection system could not apparently alter the fluorescence intensity of the probe, and only the presence of TMZ could cause the fluorescence quenching of the system (Figure 5). The experimental outcomes demonstrate that the constructed CB[7]@TTPE probe has excellent selectivity toward temozolomide and strong anti-interference ability in its practical determination. Figure 5 Specificity of the probe solution for the determination of temozolomide. The concentration of interfering substances and TMZ are both 1 mg/mL. Analysis of TMZ in serum samples In order to verify the feasibility of CB[7]@TTPE fluorescent probe in the actual fluid determination, temozolomide added in human serum samples was detected. As shown in Table 2, the average recoveries of assaying TMZ by this fluorescence technology were in the range of 102.34% to 109.07%, and the RSD values were less than 1%. The above outcomes revealed that the novel aggregation-induced emission supramolecular probe system was fast and uncomplicated to perform with satisfactory precision, accuracy and repeatability, which can be employed in the real analysis of TMZ. Table 2 Detection of TMZ in human serum samples Serum 6 6.14 ± 0.05 102.34 0.86 11 11.92 ± 0.08 108.38 0.71 15 16.36 ± 0.14 109.07 0.84 a Mean ± standard deviation (n = 5) Inhibition of cell viability In order to demonstrate the biocompatibility of supramolecular probe, the CCK-8 assay was applied to evaluate the toxic effects of temozolomide and CB[7]@TTPE probe with temozolomide on living U87 cells. As shown in Figure 6, the viability of cell growth cultured with different concentrations of the probe system was up to 85% even after prolonged incubation to high concentrations. This result suggested that the fluorescent probe owned low cytotoxicity and good biocompatibility, which was also the premise and basis for its application in biology. Figure 6 Cell viability of U87 cells incubated with different concentrations of TMZ and CB[7]@TTPE+TMZ for 24 h. TMZ imaging in cells by CB[7]@TTPE With the promising TMZ sensing property of supramolecular probe, we further researched the applicability of CB[7]@TTPE for fluorescence imaging of drugs in living cells. As described the LSCM images of bright field in Figure 7, CB[7]@TTPE successfully entered the U87 cell with good cellular morphology and located in the cytoplasm. Under the λ ex of 405 nm and 488 nm, human glioma cancer U87 cells can emit bright blue emission when they were co-incubated with CB[7]@TTPE for 2 h. When U87 cells were co-incubated with the mixture of CB[7]@TTPE and TMZ (40 μM) for 2 h, the blue emission from the cells was obviously weakened or even disappeared. Under the same observation conditions, TMZ can diminish the fluorescence of CB[7]@TTPE in human glioma cancer cells. Figure 7 Confocal fluorescence images of U87 cells incubated with CB[7]@TTPE probe without and with TMZ (40 μM) at 37°C for 2 h. Then, the fluorescence imaging of supramolecular inclusion complex in drug-resistant cells was further explored (Figure 8). When CB[7]@TTPE was incubated with T98G cells under the same conditions, the intracellular TMZ could be clearly imaged and monitored through the difference of fluorescence intensity. The significantly enhanced fluorescence observed in T98G cells compared with TMZ-sensitive U87 cells may be related to reduced cellular uptake of the drug, which was in line with the reported results that lower TMZ levels in drug-resistant cells led to reducing the quenching effect of probe system [35, 36]. The consequences of the above cell imaging assays showed that the fluorescence method was beneficial to monitor the drug resistance of temozolomide. Therefore, CB[7]@TTPE can be used as a fluorescent probe to observe and analyze temozolomide in complex cellular environment. Figure 8 a: Fluorescence images of U87 cells and T98G cells incubated with supramolecular probe with TMZ for 2 h; b: Comparison of CB[7]@TTPE fluorescence intensity in U87 cells and T98G cells. Conclusions In short, a rapid, sensitive and reliable supramolecular fluorescence sensing method based on host-guest interaction was constructed to selectively measure the level of temozolomide in actual serum samples and monitor it in living cells. CB[7]@TTPE nanoprobe fully embodies the advantages from each component (such as excellent optical property of TTPE and high molecular recognition performance of CB[7]). At the same time, the composites are stable and exhibit great solubility in aqueous solution. Through the IFE process, TMZ molecules can further quench the fluorescence of CB[7]@TTPE system with host-guest recognition function, thus a fluorescence detection platform for temozolomide was established. After optimizing various sensing conditions, the consequences indicated that AIE supramolecular probe CB[7]@TTPE possessed outstanding sensitivity, accuracy and selectivity for TMZ determination with low detection limit. The developed nanocomposite was successfully applied for the TMZ analysis of real serum samples. More importantly, the probe had wonderful biocompatibility and could enter human glioma cancer cells for TMZ imaging. It is believed that CB[7]@TTPE fluorescent probe can provide a new idea for rapid, sensitive and selective detection of temozolomide, and possesses potential application prospects in biomedical fields such as cell imaging and drug resistance monitoring. 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Biomedicine & Pharmacotherapy, 2023, 162, 114643. Google Scholar Information & Authors Information Version history V1 Version 1 02 January 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Keywords aggregation-induced emission cell imaging self-assembly supramolecular probe temozolomide detection Authors Affiliations Chenrui Jiang The First Affiliated Hospital of Ningbo University View all articles by this author Feifei Chen 0009-0002-8960-9367 The First Affiliated Hospital of Ningbo University View all articles by this author Yue Chen China Pharmaceutical University View all articles by this author Yu Sun The First Affiliated Hospital of Ningbo University View all articles by this author Hua He China Pharmaceutical University View all articles by this author Pierre Dramou China Pharmaceutical University View all articles by this author Tao Xu The First Affiliated Hospital of Ningbo University View all articles by this author Hongbin Xu 0000-0002-8406-9827 [email protected] The First Affiliated Hospital of Ningbo University View all articles by this author Metrics & Citations Metrics Article Usage 230 views 166 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Chenrui Jiang, Feifei Chen, Yue Chen, et al. Self-assembled aggregation-induced emission supramolecular probe for temozolomide detection and cell imaging. Authorea . 02 January 2025. DOI: https://doi.org/10.22541/au.173578157.70405911/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu . 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