Fluorescent Molecules Research Articles

An ESIPT-Based Fluorescent Probe for Wash-Free Cell Membrane Imaging.

The cell membrane tracking is important for studying the membrane function and diagnosing membrane-related diseases, so the development of fluorescent molecule specifically lighting up cell membrane is highly desirable. In this work, we designed and readily prepared a fluorescent dye (BOHI) that can target cell membrane. The pH-dependent emission spectra reveal that BOHI shows fluorescence based on the excited-state intramolecular proton transfer (ESIPT) under acidic and neutral conditions, while the blue-shifted emission from the phenolate anion of BOHI was observed when pH ≥ 9. Among various solvents, the weakest fluorescence emission was observed in H2O, which is favorable to wash-free imaging. The fluorescence intensity of BOHI is greatly affected by surfactants. The anionic surfactant can induce intense green fluorescence, while very weak fluorescence was observed in the solution of cationic surfactant. Hela cells images show that BOHI can specifically targets and lights up cell membrane.

STORM Super-Resolution Visualization of Self-Assembled γPFD Chaperone Ultrastructures in Methanocaldococcus jannaschii.

Gamma-prefoldin (γPFD), a unique chaperone found in the extremely thermophilic methanogen Methanocaldococcus jannaschii, self-assembles into filaments in vitro, which so far have been observed using transmission electron microscopy and cryo-electron microscopy. Utilizing three-dimensional stochastic optical reconstruction microscopy (3D-STORM), here we achieve ∼20 nm resolution by precisely locating individual fluorescent molecules, hence resolving γPFD ultrastructure both in vitro and in vivo. Through CF647 NHS ester labeling, we first demonstrate the accurate visualization of filaments and bundles with purified γPFD. Next, by implementing immunofluorescence labeling after creating a 3xFLAG-tagged γPFD strain, we successfully visualize γPFD in M. jannaschii cells. Through 3D-STORM and two-color STORM imaging with DNA, we show the widespread distribution of filamentous γPFD structures within the cell. These findings provide valuable insights into the structure and localization of γPFD, opening up possibilities for studying intriguing nanoscale components not only in archaea but also in other microorganisms.

Invisible photochromism and on-off switch based on thiophene-imidazole fused derivatives: Synthesis, properties and applications

Three novel diarylethene fluorescent molecules based on a thiophene-imidazole core have been successfully synthesized. Their photophysical properties, including photocyclization and ions response characteristics, as well as quantum chemical calculations were systematically investigated. All compounds exhibited significant invisible photochromism upon photocyclization, resulting in remarkable fluorescence “turn-off” properties. This phenomenon can be attributed to photoisomerization, specifically the transition from the open-ring isomer to the closed-ring isomer during this process, which was confirmed by 1H NMR data analysis. Moreover, AT-DPIo demonstrated high selectivity toward I− with pronounced changes in absorption from colorless to yellow and fluorescence from bright yellow to dark, enabling a limit of detection for I− at 9.75 × 10−7 mol L−1. These exceptional sensing properties make it suitable for rapid “naked-eye” detection of I− using test strips.

Immunological isolation and characterization of neuronal progenitors from human dental pulp: A laboratory-based investigation.

Dental pulp stem cells (DPSCs) contain a population of stem cells with a broad range of differentiation potentials, as well as more lineage-committed progenitors. Such heterogeneity is a significant obstacle to experimental and clinical applications. The aim of this study is to isolate and characterize a homogenous neuronal progenitor cell population from human DPSCs. Polysialylated-neural cell adhesion molecule (PSA-NCAM+) neural progenitors were isolated from the dental pulp of three independent donors using magnetic-activated cell sorting (MACS) technology. Immunofluorescent staining with a panel of neural and non-neural markers was used to characterize the magnetically isolated PSA-NCAM+ fraction. PSA-NCAM+ cells were then cultured in Neurobasal A supplemented with neurotrophic factors: dibutyryl cyclic-AMP, neurotrophin-3, B27 and N2 supplements to induce neuronal differentiation. Both PSA-NCAM+ and differentiated PSA-NCAM+ cells were used in Ca2+ imaging studies to assess the functionality of P2X3 receptors as well as membrane depolarization. PSA-NCAM+ neural progenitors were isolated from a heterogeneous population of hDPSCs using magnetic-activated cell sorting and anti-PSA-NCAM MicroBeads. Flow cytometry analysis demonstrated that immunomagnetic sorting significantly increased the purity of PSA-NCAM+ cells. Immunofluorescent staining revealed expression of pan-neuronal and mature neuronal markers, PGP9.5 and MAP2, respectively, as well as weak expression of the mature sensory markers, peripherin and islet1. ATP-induced response was mediated predominately by P2X3 receptors in both undifferentiated and differentiated cells, with a greater magnitude observed in the latter. In addition, membrane depolarizations were also detected in cells before and after differentiation when loaded with fast-voltage-responding fluorescent molecule, FluoVolt™ in response to potassium chloride. Interestingly, only differentiated PSA-NCAM+ cells were capable of spontaneous membrane oscillations. In summary, DPSCs contain a population of neuronal progenitors with enhanced neural differentiation and functional neural-like properties that can be effectively isolated with magnetic-activated cell sorting (MACS).

Solid-Emission-Tunable Squaraine with Thermal-Promoted Aggregate-State Transitions for Fast Thermal History Sensing.

Determining thermal history is crucial in many industrial processes, but reliable and sensitive organic thermal history indicators are currently absent. Herein, we report on the development of a squaraine-based fluorescent molecule, DPEA-SQ, for the detection of thermal exposure histories up to 436 K. DPEA-SQ forms multiple single crystals (DPEA-SQ-I, DPEA-SQ-II, and DPEA-SQ-III) with different conformations and aggregate-state packing modes, contributing to their different fluorescence wavelengths, lifetimes, and efficiencies. Interestingly, DPEA-SQ-I and DPEA-SQ-III undergo aggregate-state structural transitions to form the thermodynamically more stable DPEA-SQ-II, which are accompanied by changes in their fluorescence. By taking advantage of similar aggregate-state structural transformations during heating, a high-temperature thermal exposure history of up to 436 K is recorded and reflected by their fluorescence. To demonstrate the potential practical applications of DPEA-SQ, a DPEA-SQ-Powder/PDMS film is prepared and coated on an electric circuit board, which enables real-time monitoring of localized overheating by the naked eye. Additionally, the fluorescence peaks of DPEA-SQ-Powder and DPEA-SQ-Powder/PDMS films remain unchanged after storage at 373 K for 52 days, demonstrating high aggregate-state stability. The fast and reliable responses of this system make it an excellent candidate for the detection of overtemperature traces in electronic components and circuit diagnosis.

Aminonaphthalene and aminoquinoline photocages: meta-effect and photo-release of carboxylic acids and alcohols

Three series of compounds were tested as photocleavable protective groups (photocages), which are derivatives of 1,2-substituted aminonaphthalene 4, 1,3-substituted aminonaphthalene 5, or 5,6-substituted aminoquinolines 6. Their photophysical properties and photochemical reactivity in releasing alcohols and carboxylic acids were investigated. Moreover, mechanism of photochemical reaction was studied computationally and by transient absorption spectroscopy with time-resolution from femtoseconds to milliseconds. Photocages 4 and 5 are generally fluorescent molecules with high ΦF, whereas quinoline 6 is weekly fluorescent due to photoinduced charge transfer. Derivatives 4 release alcohol with low quantum yield (release of methanol from 4d is ΦR = 0.00010 ± 0.00005), but upon prolonged irradiation can be applied in organic synthesis to deprotect both aliphatic and aromatic alcohols. On the contrary, photocages 5 undergo relatively efficient release of carboxylic acids, (release of acetic acid from 5b is ΦR = 0.22 ± 0.01), in accord with the meta-effect in photochemistry, which is our main discovery that is based on the rational design of the chromophore. The photochemical reaction mechanism involves homolytic cleavage on the singlet excited state surface and formation of radicals which were detected by transient spectroscopy. The meta-effect did not promote the heterolytic cleavage as it was the case with aniline chromophores. The subsequent electron transfer delivers naphthalenmethyl carbocation and released carboxylates. The applicability of photocages 5 in organic synthesis was demonstrated on aliphatic and aromatic carboxylic acids. The potential use in biology was suggested due to their very weak photoinduced antiproliferative effect, which was essayed on three human cancer cell lines (MCF-7 breast carcinoma, HCT 116 colon carcinoma, and H460 lung carcinoma).

Dihydroxanthene-based monoamine oxidase A-activated photosensitizers for photodynamic/photothermal therapy of tumors

Small molecule photosensitizers for combined in vivo tailored cancer diagnostics and photodynamic/photothermal therapy are desperately needed. Monoamine oxidase A (MAO-A)-activated therapeutic and diagnostic compounds provide great selectivity because MAO-A can be employed as a biomarker for associated Tumors. In order to screen photosensitizers with photodynamic therapeutic potential, we have created a range of near-infrared fluorescent molecules in this work by combining dihydroxanthene parent with various heterocyclic fluorescent dyes. The NIR fluorescent diagnostic probe, DHMQ, was created by combining the screened fluorescent dye matrices with the propylamino group, which is the recognition moiety of MAO-A, based on the oxidative deamination mechanism of the enzyme. This probe has a low toxicity level and can identify MAO-A precisely. It has the ability to use fluorescence imaging on mice and cells to track MAO-A activity in real-time. It has strong phototoxicity and can produce singlet oxygen when exposed to laser light. The temperature used in photothermal imaging can get up to 50 °C, which can harm tumor cells permanently and have a positive phototherapeutic impact on tumors grown from SH-SY5Y xenograft mice. The concept of using MAO-A effectively in diseases is expanded by the MAO-A-activated diagnostic-integrated photosensitizers, which offer a new platform for in vivo cancer diagnostics and targeted anticancer treatment.

Enhancement of Upconversion Luminescence through Three-Dimensional Design of Plasma Ti3O5-Coupled Structural Domain-Limiting Effects.

Upconversion luminescence plays a crucial role in various technological applications, and among the various valence states of lanthanide elements, Ln3+ has the highest stability. The 4f orbitals of these elements are in a fully empty, semifull, or full state. This special 4f electron configuration allows them to exhibit rich discrete energy levels. However, the 4f-4f transition of Ln3+ rare earth ions itself is prohibited, resulting in a lower luminescence efficiency. This limitation greatly hinders the practical application of upconversion luminescence. In this study, we report nanostructured luminescence-enhanced substrate platforms with both semiconductive local surface plasmons and spatially confined domain effects on a single defect semiconductor substrate. By coupling NaYF4:Yb-Er nanoparticle emitters to the surface of Ti3O5 NC-arrays plasmonic nanostructures, an ultrabright luminescence with a 32-fold increase in green emission and a 40-fold increase in red emission was achieved. Furthermore, the fluorescence resonance energy transfer characteristics observed in the R6G/NaYF4/Ti3O5 NC-array composite film enable accurate detection of fluorescent molecules. The results provide an innovative and intelligent approach to enhance the upconversion luminescence intensity of rare-doped nanoparticles and develop highly sensitive molecular detection systems based on the above luminescence enhancement.

Reactive Oxygen Species-Responsive Nano-platform with Dual-Targeting and Fluorescent Lipid-Specific Imaging Capabilities for the management of Atherosclerotic Plaques

Atherosclerosis (AS), a pathological cause of cardiovascular disease, results from endothelial injury, local progressive inflammation, and excessive lipid accumulation. AS plaques rich in foam cells are prone to rupture and form thrombus, which can cause life-threatening complications. Therefore, the assessment of atherosclerotic plaque vulnerability and early intervention are crucial in reducing the mortality rates associated with cardiovascular disease. In this work, A fluorescent probe FC-TPA was synthesized, which switches the fluorescence state between protonated and non-protonated, reducing background fluorescence and enhancing signal-to-noise ratio. On this basis, FC-TPA is loaded into cyclodextrin (CD) modified with phosphatidylserine targeting peptide (PTP) and coated with hyaluronic acid (HA) to construct the intelligent responsive diagnostic nanoplatform (HA@PCFT). HA@PCFT effectively targets atherosclerotic plaques, utilizing dual targeting mechanisms. HA binds strongly to CD44, while PTP binds to phosphatidylserine, enabling nanoparticle aggregation at the lesion site. ROS acts as a smart release switch for probes. Both in vitro and in vivo evaluations confirm impressive lipid-specific fluorescence imaging capabilities of HA@PCFT nanoparticles (NPs). The detection of lipid load in atherosclerotic plaque by fluorescence imaging will aid in assessing the vulnerability of atherosclerotic plaque. STATEMENT OF SIGNIFICANCE: Currently, numerous fluorescent probes have been developed for lipid imaging. However, some challenges including inadequate water solubility, nonspecific distribution patterns, and fluorescence background interference, have greatly limited their further applications in vivo. To overcome these limitations, a fluorescent molecule has been designed and synthesized, thoroughly investigating its photophysical properties through both theoretical and experimental approaches. Interestingly, this fluorescent molecule exhibits the reversible fluorescence switching capabilities, mediated by hydrogen bonds, which effectively mitigate background fluorescence interference. Additionally, the fluorescent molecules has been successfully loaded into nanocarriers functionalized with the active targeting abilities, which has significantly improved the solubility of the fluorescent molecules and reduced their nonspecific distribution in vivo for an efficient target imaging in atherosclerosis. This study provides a valuable reference for evaluating the performance of such fluorescent dyes, and offers a promising perspective on the design of the target delivery systems for atherosclerosis.

Fabrication of chitosan-based fluorescent hydrogel membranes cross-linked with bisbenzaldehyde for efficient selective detection and adsorption of Fe2+

Chitosan, as a natural biomass material, is green, recyclable, sustainable and well biocompatible. The molecular chain is rich in active groups such as amino and hydroxyl groups, and its preparation of fluorescent probes has the advantages of biocompatibility and efficient detection performance. In this study, a bis(benzaldehyde) (BHD) fluorescent functional molecule was designed. Then a series of fluorescent chitosan-based hydrogel films (CSBHD) were prepared using chitosan as raw material and BHD as cross-linking agent. As a fluorescent probe for metal ions, CSBHD was able to efficiently detect Fe2+ with a linear correlation of fluorescence intensity in the range of 0–160 μM, and the limit of detection (LOD) was 0.55 μM. Moreover, it has excellent adsorption performance for Fe2+ ions, with a maximum adsorption capacity of 223.5 g/mg at 500 mg/L Fe2+ concentration. Finally, we characterised the structure and microscopic morphology of CSBHD films and found that CSBHD as a hydrogel film has a high cross-linking density, good water resistance, excellent thermal stability, strong resistance to swelling, and excellent stability in cycling tests. Hence, it has great potential for application in adsorption and detection of Fe2+ ions. It also provides a good strategy for the application of chitosan based fluorescent probe materials in environmental monitoring and heavy metal ion adsorption.

Rigid symmetric Rhodamine fluorescent dyes as fast selective detection of aluminum and iron ions applied to cell imaging

Fluorescent probe molecules have the advantages of good selectivity, high sensitivity, and easy operation, which have attracted much attention. In this paper, a novel symmetric rhodamine fluorescent probe with structural rigidity was synthesized by a simple reaction using rhodamine 6G as raw material. And it was characterized by NMR and electrospray spray mass spectrometry. Single crystal was prepared by solvent evaporation method, which was the first single crystal of the compound reported domestically and internationally. Special spiral ring structures were observed inside the crystal. Among different competing cations, the probe exhibited high selectivity and good sensitivity towards Al3+ and Fe3+ metal ions. The limits of detection were 0.76 nM and 0.49 nM, respectively. In the presence of Al3+ and Fe3+, the solution changed from colorless to pink. At the same time, there was a significant shift in the UV-vis spectrum and fluorescence emission spectrum. The UV-vis absorption peak appeared at 530 nm, while the emission at 556 nm was turned on. In addition, biological experiments have shown that the RP probe has good membrane permeability and has the potential to detect Al3+ and Fe3+ in Hela cells.

NIR-Activated Water-soluble Molecular Probe Based on Cu(ii) Ion Quenching Mechanisms and Substituted Strategies: For In vitro/In vivo H2S Detection

Human health, environmental contamination, and food safety are tightly associated with hydrogen sulfide (H2S). The safe and highly sensitive monitoring of H2S in tissues, organs, and blood has presented ongoing challenges. Herein, a novel near-infrared (NIR)-activated water-soluble molecular probe, denoted as IR820-TPY-Cu was developed for detecting hydrogen sulfide. This probe is built upon the fluorescent molecule IR820, a derivative of Indocyanine green (ICG). Upon reacting with H2S, IR820-TPY-Cu exhibits active fluorescence (670-750nm), which is attributed to the Cu (II) ion quenching mechanism and substitution strategy. IR820-TPY-Cu exhibits selectively activated near-infrared fluorescence signal changes, a low detection limit of 0.114μM, and a rapid in vitro response (3min) for H2S detection. Moreover, the biocompatible probe IR820-TPY-Cu not only quantitatively monitors H2S in environmental water and various food specimens (red wine, beer, meat, milk, and sweet potato) through colorimetric sensing but also successfully images endogenous and exogenous H2S in living cells, zebrafish, and mice. This research provides a promising approach for monitoring H2S in environmental contamination, ensuring food safety, and studying living systems.

Design and Preparation of Multicolor Long‐Lifetime Fluorescent Material Based on Charge‐Separated States

Design and Preparation of Multicolor Long‐Lifetime Fluorescent Material Based on Charge‐Separated States

Light-Driven Mitochondrion-to-Nucleus DNA Cascade Fluorescence Imaging and Enhanced Cancer Cell Photoablation.

Nucleic acids are mainly found in the mitochondria and nuclei of cells. Detecting nucleic acids in the mitochondrion and nucleus in cascade mode is crucial for understanding diverse biological processes. This study introduces a novel nucleic acid-based fluorescent styrene dye (SPP) that exhibits light-driven cascade migration from the mitochondrion to the nucleus. By introducing N-arylpyridine on one side of the styrene dye skeleton and a bis(2-ethylsulfanyl-ethy)-amino unit on the other side, we found that SPP exhibits excellent DNA specificity (16-fold, FDNA/Ffree) and a stronger binding force to nuclear DNA (-5.09 kcal/mol) than to mitochondrial DNA (-2.59 kcal/mol). SPP initially accumulates in the mitochondrion and then migrates to the nucleus within 10 s under light irradiation. By tracking the damage to nucleic acids in apoptotic cells, SPP allows the successful visualization of the differences between apoptosis and ferroptosis. Finally, a triphenylamine segment with photodynamic effects was incorporated into SPP to form a photosensitizer (MTPA-SPP), which targets the mitochondria for photosensitization and then migrates to the nucleus under light irradiation for enhanced photodynamic cancer cell treatment. This innovative nucleic acid-based fluorescent molecule with light-triggered mitochondrion-to-nucleus migration ability provides a feasible approach for the in situ identification of nucleic acids, monitoring of subcellular physiological events, and efficient photodynamic therapy.

Photorewriting, Time-Resolved Encryption, and Unclonable Anticounterfeiting with Artificial Intelligence Authentication via a Reversible Photoswitchable System.

In the fields of photolithographic patterning, optical anticounterfeiting, and information encryption, reversible photochromic materials with solid-state fluorescence are emerging as a potential class of systems. A design strategy for reversible photochromic materials has been proposed and synthesized through the introduction of photoactive thiophene groups into the molecular backbone of aryl vinyls, compounds with unique aggregation-induced emission properties, and solid-state reversible photocontrollable fluorescence and color-changing properties. This work develops novel photochromic inks, films, and cellulose hydrogels for enhancing the security of information encryption and anticounterfeiting technologies. They achieve rapid and reversible color change under ultraviolet light irradiation. Dependent upon the rate of color change, higher levels of time-resolved security can be achieved. This feature is important for enhancing the confidentiality of encrypted information and the reliability of security labels. Color-changing cellulose hydrogels, inks, and films consisting of three photochromic fluorescent molecules have quick photoactivity, great photoreversibility and photostability, and good processability, making them ideal for time-delayed anticounterfeiting and smart encryption. Furthermore, specialized algorithms are used to construct convolutional neural networks, and image analysis is performed on these systems, thus solving the current problem of the time-consuming information decryption process. This artificial intelligence method offers new opportunities for enhanced data encryption.

Visible Light-Driven Metal-free and Photocatalyst-free Intramolecular Cyclization of Chalcones to Access Phenanthrenes

We report a visible light-driven metal-free and photocatalyst-free protocol to the synthesis of phenanthrenes via intramolecular cyclization of chalcones. The transformation proceeds via light irradiation and base- and oxygen-based promotion and enables the generation of a series of phenanthrenes. The further functionalization of the as-synthesized phenanthrene to afford fluorescent molecule was explored.

Pyrrole-substituted difluoroboron β-diketonate dyes: Facile synthesis and multi-state optical properties

Difluoroboron β-diketonate complexes (BF2bdks) are classic fluorescent molecules that have been applied in luminescent materials, oxygen and mechanical sensors, and most investigations concentrate on BF2bdks with varying aromatic groups without heteroatoms. In this work, a new molecular platform BF2bdks with heterocycle-substituted is prepared in one pot, and six unsymmetric π-expanded BF2bdks are synthesized via condensation reaction. B-1∼6 exhibit comparable fluorescence properties in solution, aggregated and solid states. B-3 (683 nm, Φf = 13.1 %) and B-5 (700 nm, Φf = 2.1 %) show near infra-red (NIR) emission with moderate fluorescence intensity in solution state. Solid emission of B, B-1-3 and B-5 are detected with fluorescence quantum yield from 12.8 % to 25.6 %, and B-2, B-3 and B-5 show AIE phenomenon in aggregated state. The results are supported by crystal analysis and DFT calculation in some extent. In a word, this work enriches the heterocycle-substituted BF2bdks system. B-3 and B-5 with moderate intensity NIR emission in solution, aggregated and solid states at the same time, providing a simple preparation method for multi-states luminescent dyes.

AIE-labeled fluorescent polystyrene nanoplastics for quantitative analysis in macrophages uptake

Nanoplastics have attracted significant attention due to their harmful effects on the environment and organisms. However, accurately detecting and quantifying nanoplastics that are ingested by organisms poses a challenge. Conventional organic fluorescent-labeled nanoplastics have reduced labeling efficiency due to aggregation-caused quenching (ACQ), leading to artifacts caused by leakage of fluorescent molecules. To address these limitations, this study utilized AIE fluorescence for internally labeling polystyrene nanoparticles. AIE-active molecules do not emit fluorescence in dilute solutions, thus preventing accuracy issues associated with fluorescence leakage. Three model plastics (TPE@PS) of varying particle sizes were synthesized via the swelling method. The preparation parameters were optimized, followed by stability tests conducted on TPE@PS. Fluorescence microscopy imaging revealed significant uptake of TPE@PS by RAW246.7 cells. A successful standard curve of fluorescence intensity-concentration was established successfully and utilized to accurately quantify the amount of TPE@PS that was ingested by RAW246.7 cells. The results indicate a consistent increase with increasing concentration in TPE@PS uptake across all particle sizes during the fixed exposure duration of 48 h. Furthermore, RAW264.7 cells exhibited a higher propensity to uptake smaller TPE@PS particles. Flow cytometry was also employed to validated the suitability of TPE@PS as a model nanoplastic for tracing purposes.

MINFLUX nanoscopy: Visualising biological matter at the nanoscale level

Since its introduction in 2017, MINFLUX nanoscopy has shown that it can visualise fluorescent molecules with an exceptional localisation precision of a few nanometres. In this overview, we provide a brief insight into technical implementations, fluorescent marker developments and biological studies that have been conducted in connection with MINFLUX imaging and tracking. We also formulate ideas on how MINFLUX nanoscopy and derived technologies could influence bioimaging in the future. This insight is intended as a general starting point for an audience looking for a brief overview of MINFLUX nanoscopy from theory to application.

In situ quantitative mapping of coding single nucleotide polymorphism on mRNA inside cells by SERS-fluorescence dual-mode probe

Surface Enhanced Raman Scattering (SERS) has potential in nucleic acid detection, but its ability on single nucleotide polymorphisms (SNPs) detection in the early diagnosis of diseases is still limited. In fact, there is few research about in situ detection of coding SNPs which are carried on mRNA. In this paper, we developed SERS-Fluorescence Dual-mode probes to achieve in situ quantitative mapping of intracellular cSNP. The probes were based on Au@Ag core-shell structure that possessed both excellent biocompatibility and localized surface plasmon resonance. Fluorescence can provide a rapid visualization of intracellular cSNP, while SERS can achieve its in situ quantification. The quantitation was conducted by comparing the SERS signals of the reporting molecule p-mercaptobenzoic acid and fluorescent molecule FAM, the detecting limitation of which can be achieved as low as 1.4 fM. This ratiometric detection is more accurate than traditional analysis based on signal intensity directly. It successfully achieved intracellular imaging of the G12D mutation site of KRAS gene in A549 cells, which can help to get the distribution of cSNP in single cell more clearly. And it also demonstrated the good mapping ability for biomolecules (1003 cm−1 for phenylalanine) simultaneously. In conclusion, this kind of probe could be used as a potential tool to monitor cSNP as well as biomolecules in multiple biological processes, which could provide reference for disease prediction.