CN115825025A - Application of chiral carbon quantum dots - Google Patents
Application of chiral carbon quantum dots Download PDFInfo
- Publication number
- CN115825025A CN115825025A CN202211248927.8A CN202211248927A CN115825025A CN 115825025 A CN115825025 A CN 115825025A CN 202211248927 A CN202211248927 A CN 202211248927A CN 115825025 A CN115825025 A CN 115825025A
- Authority
- CN
- China
- Prior art keywords
- carbon quantum
- chiral carbon
- bacteria
- quantum dots
- gram
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses application of chiral carbon quantum dots as a bacteria and/or fungus imaging material, and also discloses a method for distinguishing gram-positive bacteria/fungi and gram-negative bacteria. The chiral carbon quantum dot (the levorotatory chiral carbon quantum dot or the dextrorotatory chiral carbon quantum dot) has the advantages of low toxicity, good biocompatibility, photobleaching resistance and stable fluorescence property, has chiral property, has fluorescence emission performance depending on excitation wavelength, and is suitable to be used as an imaging material of bacteria and/or fungi. By adopting the chiral quantum dots as imaging materials, the method can realize the differentiation of gram-positive bacteria and fungi and the differentiation of gram-positive bacteria and gram-negative bacteria.
Description
Technical Field
The invention relates to a new application of chiral carbon quantum dots, in particular to an application of the chiral carbon quantum dots as fungus/bacteria imaging materials.
Background
Carbon Quantum Dots (CQDs), also called Carbon Dots (CDs) or Carbon nanodots, are spherical fluorescent nanoparticles with Carbon as a framework structure, good dispersibility and particle size less than 10nm, and are a novel zero-dimensional Carbon-based nanomaterial discovered after fullerene, carbon nanotube and graphene. Due to the characteristics of nano-size effect, adjustable photoluminescence property, low toxicity, good biocompatibility, easy surface modification and the like, the carbon quantum dots are widely researched and applied in the fields of biological detection, fluorescence imaging, catalysis, tumor resistance and the like.
On the other hand, bacterial infections are one of the biggest challenges facing the world, and rapid diagnosis of bacterial infections is of crucial importance for clinical treatment. The standard method for identifying unknown bacteria is gram staining, which classifies bacterial species into two categories: gram positive and gram negative. However, this method has some disadvantages, such as cumbersome procedures and the easy generation of false positive results. Quantitative real-time PCR (qPCR) has been considered a highly sensitive bacterial species identification technique, but this technique is too expensive for many places in developing countries, which greatly limits its practical application. Therefore, the invention of rapid bacterial infection diagnosis and simple and efficient bacterial identification technology is very urgent.
Disclosure of Invention
The invention aims to solve the technical problem of providing a new application of chiral carbon quantum dots.
The technical problem to be solved by the present invention is to provide a method for differentiating gram-positive bacteria/fungi from gram-negative bacteria.
In order to solve the technical problems, the invention provides an application of chiral carbon quantum dots as a bacterial and/or fungal imaging material.
Specifically, the inventor finds that the chiral carbon quantum dots (the left-handed chiral carbon quantum dots or the right-handed chiral carbon quantum dots) have good chiral properties, stable fluorescence performance and fluorescence emission characteristics dependent on laser wavelength through a large amount of research. In addition, the chiral carbon quantum dots have low toxicity, good biocompatibility and photobleaching resistance. Therefore, the method has good application prospect in the aspect of imaging materials of bacteria and/or fungi.
Specifically, in an embodiment of the present invention, a preparation method of the chiral carbon quantum dot comprises: sodium hydroxide, L-cysteine or D-cysteine and water are mixed according to the proportion of (0.1-1): (3-7): (120-200), reacting for 12-20 h at 100-140 ℃, and centrifuging and dialyzing the obtained reaction solution to obtain the chiral carbon quantum dots.
Wherein, the levo-cysteine or dextro-cysteine is used as a chiral source and a carbon source, sodium hydroxide is used as a catalyst for the carbonization polymerization reaction, water is used as a dispersing agent, and the hydrothermal reaction is carried out after a reaction system is formed. Centrifuging after hydrothermal reaction, filtering the obtained supernatant by using a cylindrical filter membrane with the aperture of 0.2-0.3 mu m, dialyzing for 12-36 h by using a dialysis belt with the molecular weight cutoff of 500-1000 Da (taking water as dialysis external liquid, replacing the dialysis external liquid every 4 h) to obtain a chiral carbon quantum dot aqueous solution, and then carrying out vacuum freeze drying at-100 to-50 ℃ for 24-48 h to obtain a finished product of the chiral carbon quantum dot.
The chiral carbon quantum dots prepared by the method are uniformly dispersed, and the average particle size is 4-5.5 nm. More specifically, the average grain diameter of the levorotatory chiral carbon quantum dots (L-CDs) is 5-5.5 nm, and the lattice spacing is 0.2-0.25 nm; the average grain diameter of the right-handed chiral carbon quantum dots (D-CDs) is 4-5 nm, the lattice spacing is 0.3-0.35 nm, and both have good crystal structures. Furthermore, the chiral carbon quantum dots prepared by the method have stable fluorescence performance, which lays a good foundation for the chiral carbon quantum dots as imaging materials of bacteria and/or fungi.
The traditional semiconductor quantum dots have high toxicity and poor light stability, and are difficult to be applied as imaging materials of bacteria and/or fungi. In addition, the chiral amino acid functionalized modified graphene quantum dot can selectively kill bacteria without toxicity to mammalian cells, and can perform biological imaging on escherichia coli, staphylococcus aureus and Hela cells within 3 h. However, the carbon quantum dot cannot effectively distinguish gram-negative bacteria from gram-positive bacteria within a short time (30 min), and cannot distinguish gram-negative bacteria from fungi, so that the carbon quantum dot cannot be used as an imaging material for gram-positive bacteria/fungi.
After the chiral carbon quantum dots and the bacteria/fungi are incubated for about 30min, gram-positive bacteria and fungi can be well dyed, and fluorescence imaging is not performed on the gram-negative bacteria, so that the aim of distinguishing the gram-negative bacteria from the gram-positive bacteria/fungi is fulfilled. In addition, the chiral carbon quantum dot can realize blue, green and red three-color fluorescence imaging.
Further, in one embodiment of the present invention, the bacteria are gram-positive bacteria. Specifically, the inventor finds that the chiral carbon quantum dots can realize good staining of gram-positive bacteria through a large number of experiments, so that the chiral carbon quantum dots can display blue, green and red fluorescence under the excitation of three kinds of laser, and further achieve the purpose of identifying the chiral carbon quantum dots. Preferably, the gram-positive bacteria are staphylococcus aureus (s. Aureus, ATCC 25923), methicillin-resistant staphylococcus aureus (MRSA, ATCC 43300), enterococcus faecalis (e.faecalis, ATCC 29212), bacillus subtilis (b.subtilis, CMCC (B) 63501).
Further, in one embodiment of the present invention, the fungi are candida albicans (c.albicans, ATCC 10231), candida parapsilosis (c.parapsilosis, ATCC 22019), but are not limited thereto.
Correspondingly, the invention also discloses a method for distinguishing gram-positive bacteria/fungi and gram-negative bacteria, which comprises the steps of incubating the bacteria to be detected and the chiral carbon quantum dots together and then detecting.
Wherein, the chiral carbon quantum dots can be left-handed chiral carbon quantum dots (L-CDs) or right-handed chiral carbon quantum dots (D-CDs).
Wherein the common incubation time is more than or equal to 30min, and if the incubation time is less than 30min, the fluorescence observed in gram-positive bacteria/fungi is weaker. Preferably, the incubation time is 30-40 min, and the fluorescence is obvious, the detection accuracy is high, and the detection efficiency is high.
The detection is performed by flow cytometry or laser Confocal (CLSM) microscopy, but is not limited thereto. Preferably, detection is by laser Confocal (CLSM) microscopy; the detection wavelengths are 405nm, 488nm and 552nm respectively. Further, when laser confocal microscopy (CLSM) is adopted for detection, if the bacteria to be detected show blue, green and red fluorescence under excitation of three excitation wavelengths of 405nm, 488nm and 552nm, the bacteria to be detected are gram-positive bacteria or fungi; otherwise, the bacteria to be detected are gram-negative bacteria.
Preferably, in one embodiment of the present invention, the gram-negative bacteria are escherichia coli, proteus mirabilis and/or salmonella; the gram-positive bacteria are staphylococcus aureus, methicillin-resistant staphylococcus aureus, enterococcus faecalis and/or bacillus subtilis.
Preferably, in one embodiment of the present invention, the fungus is candida albicans and/or candida parapsilosis;
the gram-negative bacteria are escherichia coli, proteus mirabilis and/or salmonella.
The implementation of the invention has the following beneficial effects:
the invention provides an application of chiral quantum dots as an imaging material of bacteria and/or fungi. Specifically, the chiral carbon quantum dot (the left-handed chiral carbon quantum dot or the right-handed chiral carbon quantum dot) has the advantages of low toxicity, good biocompatibility, photobleaching resistance and stable fluorescence performance, has chiral properties, has fluorescence emission depending on laser wavelength, and is suitable for serving as an imaging material of bacteria and/or fungi. By adopting the chiral quantum dot as an imaging material, the detection of gram-positive bacteria and fungi and the differentiation of the gram-positive bacteria and the gram-negative bacteria can be achieved.
Drawings
FIG. 1 is a diagram of structural characterization and photophysical properties of chiral carbon quantum dots in example 1 and example 2; wherein A is a circular dichroism spectrogram of chiral carbon quantum dots (D-CDs, L-CDs); b is a Zeta potential diagram of chiral carbon quantum dots (D-CDs and L-CDs); c is a transmission electron microscope topography and a particle size distribution diagram of the right-handed chiral carbon quantum dots (D-CDs); d is a high-resolution transmission electron microscope and a crystal lattice morphology diagram of the right-handed chiral carbon quantum dots (D-CDs); e is a fluorescence spectrum of right-handed chiral carbon quantum dots (D-CDs); f is a fluorescence spectrum of right-handed carbon quantum dots (D-CDs) in tetrahydrofuran-water mixtures with different proportions; g is a line graph of the change of the fluorescence intensity of right-handed carbon quantum dots (D-CDs) in tetrahydrofuran-water mixtures with different proportions; h is a transmission electron microscope topography of right-handed carbon quantum dots (D-CDs) in a 60% (v: v) tetrahydrofuran-water mixture;
fig. 2 is a diagram of structural characterization and photophysical properties of the chiral carbon quantum dots in example 1 and example 2; wherein A is an ultraviolet-visible absorption spectrogram of chiral carbon quantum dots (D-CDs, L-CDs); b is a Fourier transform infrared spectrogram of chiral carbon quantum dots (D-CDs and L-CDs); c is a transmission electron microscope topography and a particle size distribution diagram of the levorotatory chiral carbon quantum dots (L-CDs); d is a high-resolution transmission electron microscope and a crystal lattice topography of the levorotatory chiral carbon quantum dots (L-CDs); e is a fluorescence spectrum of the levorotatory chiral carbon quantum dots (L-CDs); f is a fluorescence spectrum of the L-chiral carbon quantum dots (L-CDs) in the tetrahydrofuran-water mixture with different proportions; g is a line graph of the change of the fluorescence intensity of the L-chiral carbon quantum dots (L-CDs) in the tetrahydrofuran-water mixture with different proportions; h is a transmission electron microscope topography of levorotatory chiral carbon quantum dots (L-CDs) in a 70% (v: v) tetrahydrofuran-water mixture;
FIG. 3 is a diagram showing the results of laser Confocal (CLSM) microscope images and flow cytometry detection of chiral carbon quantum dots and gram-positive bacteria; wherein A is right-handed chiral carbon quantum dots (D-CDs), and B is left-handed chiral carbon quantum dots (L-CDs);
FIG. 4 is a diagram showing the results of laser Confocal (CLSM) microscope images and flow cytometry detection of chiral carbon quantum dots and gram-negative bacteria; wherein A is right-handed chiral carbon quantum dots (D-CDs), and B is left-handed chiral carbon quantum dots (L-CDs);
FIG. 5 is a diagram of a laser Confocal (CLSM) microscope image of chiral carbon quantum dots and fungi and a flow cytometer detection result; wherein A is right-handed chiral carbon quantum dots (D-CDs), and B is left-handed chiral carbon quantum dots (L-CDs).
Detailed Description
To make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying drawings.
Example 1 preparation and characterization of L-chiral carbon Quantum dots (L-CDs)
The embodiment provides a preparation method of a levorotatory chiral carbon quantum dot, which comprises the following specific steps:
(1) Precisely weighing 0.06g of sodium hydroxide in 15mL of ultrapure water, stirring and dissolving by using a glass rod, adding 0.5g of L-cysteine, stirring and dissolving, and performing ultrasonic treatment for 10min until the sample is fully and uniformly mixed;
(2) Transferring the dispersed solution into a 50mL polytetrafluoroethylene lining, and filling the lining into a steel sleeve of a high-pressure reaction kettle;
(3) Placing the reaction kettle in an electric heating forced air drying box, and carrying out hydrothermal reaction for 16h at 120 ℃;
(4) After the reaction is finished, cooling the reaction kettle to room temperature, centrifuging the obtained orange-red suspension at 10000rpm for 10min, and collecting supernatant;
(5) Filtering the supernatant with a 0.22 μm cylindrical filter membrane, transferring the obtained filtrate into a MW =1000Da dialysis bag, and dialyzing in deionized water for 24h, wherein the dialysis external liquid is replaced every 4 h;
(6) After the dialysis process is finished, collecting the yellow solution in the dialysis bag to obtain pure left-handed chiral carbon quantum dot aqueous solution;
(7) And (3) freeze-drying the yellow carbon quantum dot solution for 48h at-60 ℃ by using a vacuum freeze dryer to obtain brown powder, namely the levorotatory chiral carbon quantum dot, and storing the brown powder at 4 ℃ for later use.
Example 2 preparation and characterization of right-handed chiral carbon Quantum dots (D-CDs)
The embodiment provides a preparation method of a dextrorotatory chiral carbon quantum dot, which specifically comprises the following steps:
(1) Precisely weighing 0.06g of sodium hydroxide in 15mL of ultrapure water, stirring and dissolving by using a glass rod, adding 0.5g of dextro-cysteine, stirring and dissolving, and performing ultrasonic treatment for 10min until the sample is fully and uniformly mixed;
(2) Transferring the dispersed solution into a 50mL polytetrafluoroethylene lining, and filling the lining into a steel sleeve of a high-pressure reaction kettle;
(3) Placing the reaction kettle in an electric heating forced air drying box, and carrying out hydrothermal reaction for 16h at 120 ℃;
(4) After the reaction is finished, cooling the reaction kettle to room temperature, centrifuging the obtained orange-red suspension at 10000rpm for 10min, and collecting supernatant;
(5) Filtering the supernatant with a 0.22 μm cylindrical filter membrane, transferring the obtained filtrate into a MW =1000Da dialysis bag, and dialyzing in deionized water for 24h, wherein the dialysis external liquid is replaced every 4 h;
(6) After the dialysis process is finished, collecting the yellow solution in the dialysis bag to obtain a pure right-handed chiral carbon quantum dot aqueous solution;
(7) And (3) freeze-drying the yellow carbon quantum dot solution for 48h at-60 ℃ by using a vacuum freeze dryer to obtain brown powder, namely the right-handed chiral carbon quantum dot, and storing the brown powder at 4 ℃ for later use.
The levorotatory carbon quantum dots (L-CDs) and dextrorotatory carbon quantum dots (D-CDs) obtained in the embodiments 1 and 2 are tested, and the specific test method is as follows:
1) 2mL of chiral carbon quantum dot solution with the concentration of 0.01mg/mL is absorbed by a pipette and placed in a quartz cuvette, and a CD signal is collected in a circular dichroism spectrometer; placing the sample in an ultraviolet spectrophotometer to test the ultraviolet absorption spectrum of 200-800 nm; and testing the fluorescence emission spectrum of the chiral carbon quantum dots under the excitation wavelength of 280-500 nm.
2) Preparing the chiral carbon quantum dots into 0.5mg/mL solution by using ultrapure water, performing ultrasonic dispersion treatment for 20min, sucking 1mL of solution by using a liquid-transferring gun, injecting the solution into a Malvern Zeta potential vessel, and performing Zeta potential test on the chiral carbon quantum dots by using a Malvern laser particle size analyzer ZS 90.
3) Preparing 1mg/mL solution of chiral carbon quantum dots by using ultrapure water, performing ultrasonic dispersion treatment for 20min, sucking 10 mu L of the dispersed solution by using a liquid transfer gun, dropwise adding the solution into a copper mesh special for a 300-mesh transmission electron microscope, naturally drying the solution for 24h at room temperature, and observing the solution under the transmission electron microscope at 200kV and 300kV acceleration voltage.
4) Accurately weighing the chiral carbon quantum dot powder and dry potassium bromide according to the mass ratio of 1.
5) Aggregation-induced emission effect test: preparing a mixture of tetrahydrofuran and water with the volume ratio v: v =0% -100%, adding chiral carbon quantum dots with the final concentration of 10 mu g/mL, and testing the fluorescence spectrum at the excitation wavelength of 340 nm.
The specific test results are shown in fig. 1. Specifically, the circular dichroism spectrum of fig. 1A shows that L-CDs and D-CDs present opposite and symmetrical chiral signals, which indicates that the carbon quantum dots successfully inherit the chiral property of the L/D cysteine raw material; FIG. 1B shows that chiral L/D-CDs have similar Zeta potential values; the Transmission Electron Microscope (TEM) test result of C, D in FIG. 1 shows that the prepared chiral carbon quantum dots are spherical and uniformly dispersed, and have an average particle size of 4-5.5 nm, wherein the average particle size of L-CDs is 5.2nm, the average particle size of D-CDs is 4.3nm, and the prepared chiral carbon quantum dots have a good crystal structure and a lattice spacing of 0.22 or 0.33nm; the fluorescence spectra in fig. 1E and 2E show that the chiral carbon quantum dots have fluorescence emission characteristics dependent on the excitation wavelength, and have maximum emission under excitation at 340 nm; figure 2A shows uv spectra showing the presence of two absorption peaks at 275nm and 320nm for chiral carbon quantum dots, respectively, due to C = C pi-pi transition and C = O n-pi transition; FIG. 2B is a Fourier transform infrared spectrogram at 3400cm -1 Has a broad peak and is at 3207cm -1 Due to O-H and N-H stretching vibrations, 2975 and 2920cm -1 The peak at (A) is derived from C-H and is at 1610cm -1 And 1135cm -1 Respectively, may be related to C = O and C-O stretching vibrations at 1380cm -1 The peaks of (A) may be derived from C-N, N-H and COO - . Wherein, 2550 and 1190cm -1 The peaks at (A) are related to the presence of S, respectively due to-S-H and C-S, and the peaks at these two positions of L/D-CDs are not evident in comparison to the starting cysteine, probably due to polymerization during hydrothermal processing. According to infrared spectrum, the chiral carbon quantum dots have-COOH, -OH and-NH 2 And the like, which is related to the good water solubility characteristic of the carbon quantum dots. In the aggregation-induced emission behavior test of the chiral carbon quantum dots, fig. 1F shows that the fluorescence intensity of the carbon quantum dots increases with the volume ratio of THF within a certain range, and D-CDs and L-CDs are respectively in THF: h 2 Maximum fluorescence intensity reached at O =60% and 70% (v: v). Further observing by a transmission electron microscope to obtain the chiral carbon quantum dots in THF-H 2 Aggregated particles with the diameter of 20-30nm are formed in an O two-component system, and the fact that the L/D-CDs have aggregation induction effect is verified.
Example 3 confocal imaging testing of chiral carbon quantum dots and bacteria
The test strains used were: gram-positive bacteria: staphylococcus aureus (s.aureus, ATCC 25923), methicillin-resistant staphylococcus aureus (MRSA, ATCC 43300), enterococcus faecalis (e.faecalis, ATCC 29212), bacillus subtilis (b.subtilis, CMCC (B) 63501). Gram-negative bacteria: escherichia coli (e.coli, ATCC 25922), proteus mirabilis (p.mirabilis, pro10 (AO 250)), salmonella (s.typhimurium, ATCC 14028). Fungi: candida albicans (c.albicans, ATCC 10231), candida parapsilosis (c.parapsilosis, ATCC 22019). Single bacterial colonies were picked with the inoculating loop in 5mL LB broth, single Candida albicans colonies were picked in Staphylococcus saxifragi medium, and cultured overnight with shaking at 37 ℃ and 200rpm for 18h. The bacteria and fungi were collected by centrifugation at 5000rpm for 3min, resuspended in sterile PBS buffer (10 mM, pH = 7.4), and the bacterial concentration was adjusted to 1X 10 6 ～10 7 CFU/mL, bacteria, fungi and L/D-CDs (final concentration 50. Mu.g/mL) were incubated at 37 ℃ for 30min on a shaker at 200 rpm. After incubation, wash 2 times with sterile PBS, finally resuspend with 1mL PBS and vortex mix. Pipette 10. Mu.L of the pipette tip into a clean slide, cover the slide carefully with tweezers, and observe the slide under a confocal laser microscope with a 100-fold oil lens.
The confocal images of fig. 3A, 3B, 5A, and 5B show that after being treated with chiral carbon quantum dots for 30min, four gram-positive bacteria and two fungi respectively show blue, green, and red fluorescence under laser with three excitation wavelengths of 405, 488, and 552nm. Fig. 4A, B shows that no significant fluorescence of carbon quantum dots was observed for any of the three gram-negative bacteria tested, demonstrating that chiral carbon quantum dots enter gram-positive bacteria/fungi in a short time and cannot enter gram-negative bacteria. Gram-positive bacteria/fungi can be better distinguished from gram-negative bacteria.
Example 4 flow cytometry testing of chiral carbon quantum dots and bacteria
Single colonies of the test strains were picked with the inoculating loop in 5mL LB broth, single colonies of Candida albicans were picked in a Saccharomycotina medium, and cultured with shaking at 37 ℃ and 200rpm for 18h, respectively. The bacteria/fungi were collected by centrifugation at 5000rpm for 3min, resuspended in sterilized PBS buffer (10mM, pH = 7.4), and the bacterial concentration was adjusted to 1X 10 6 ～10 7 CFU/mL, bacteria/fungi were incubated with L/D-CDs (final concentration 50. Mu.g/mL) at 37 ℃ for 30min on a shaker at 200rpm, with PBS as a control. After incubation, washing with sterilized PBS for 2 times, finally resuspending with 1mL PBS, vortexing and mixing uniformly, and quantitatively detecting the fluorescence intensity of the chiral carbon quantum dots in the bacteria by using a flow cytometer.
The results are shown in FIGS. 3, 4 and 5. Compared with the control group, the flow cytometry can detect the obvious increase of the fluorescence signal in gram-positive bacteria/fungi, and three kinds of gram-negative bacteria can not detect the obvious fluorescence signal. The chiral carbon quantum dots can be proved to be capable of better distinguishing gram-positive bacteria/fungi from gram-negative bacteria.
While the foregoing is directed to the preferred embodiment of the present invention, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention.
Claims (10)
1. Application of chiral carbon quantum dots as bacterial and/or fungal imaging materials.
2. The use of claim 1, wherein the bacteria are gram positive bacteria.
3. The use of claim 1 or 2, wherein the bacteria are staphylococcus aureus, methicillin-resistant staphylococcus aureus, enterococcus faecalis, and/or bacillus subtilis;
the fungi are Candida albicans and/or Candida parapsilosis.
4. A method for distinguishing gram-positive bacteria and gram-negative bacteria is characterized by comprising the steps of incubating bacteria to be detected and chiral carbon quantum dots together, and then detecting; wherein the incubation time is more than or equal to 30min;
after incubation, blue, green, and red fluorescence emitted from chiral carbon quantum dots was observed in gram-positive bacteria and not in gram-negative bacteria at excitation wavelengths of 405nm, 488nm, and 552nm.
5. The method of claim 4, wherein the gram-negative bacteria are Escherichia coli, proteus mirabilis, and/or Salmonella;
the gram-positive bacteria are staphylococcus aureus, methicillin-resistant staphylococcus aureus, enterococcus faecalis and/or bacillus subtilis.
6. A method for distinguishing fungi from gram-negative bacteria is characterized by comprising the steps of incubating bacteria to be detected and chiral carbon quantum dots together, and then detecting; wherein the incubation time is more than or equal to 30min;
after incubation, blue, green, and red fluorescence emitted from chiral carbon quantum dots was observed in fungi at excitation wavelengths of 405nm, 488nm, and 552nm, and no fluorescence was observed in gram-negative bacteria.
7. The method of claim 6, wherein the fungus is Candida albicans and/or Candida parapsilosis;
the gram-negative bacteria are escherichia coli, proteus mirabilis and/or salmonella.
8. The method of claim 4 or 6, comprising:
resuspending the bacteria to be detected with sterilized PBS, wherein the concentration of the bacteria to be detected is 1 × 10 6 ～1×10 7 CFU/mL；
Mixing the chiral carbon quantum dots with the resuspended bacteria to be detected, and incubating for 10-30 min in a shaking table with the temperature of 30-45 ℃ and the rpm of 180-250; wherein the concentration of the chiral carbon quantum dots is 40-70 mu g/mL;
and washing the incubated product for 1-3 times by using sterilized PBS, finally re-suspending by using sterilized PBS, and detecting by using a flow cytometer or a laser confocal microscope after uniformly vortexing.
9. The application of claim 1, wherein the chiral carbon quantum dot is prepared by a method comprising the following steps:
sodium hydroxide, L-cysteine or D-cysteine and water are mixed according to the proportion of (0.1-1): (3-7): (120-200), reacting for 12-20 h at 100-140 ℃, and centrifuging and dialyzing the obtained reaction solution to obtain the chiral carbon quantum dots.
10. The method of claim 4 or 6, wherein the chiral carbon quantum dot is prepared by a method comprising:
sodium hydroxide, L-cysteine or D-cysteine and water are mixed according to the proportion of (0.1-1): (3-7): (120-200), reacting for 12-20 h at 100-140 ℃, and centrifuging and dialyzing the obtained reaction solution to obtain the chiral carbon quantum dots.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211248927.8A CN115825025A (en) | 2022-10-12 | 2022-10-12 | Application of chiral carbon quantum dots |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211248927.8A CN115825025A (en) | 2022-10-12 | 2022-10-12 | Application of chiral carbon quantum dots |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115825025A true CN115825025A (en) | 2023-03-21 |
Family
ID=85524676
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211248927.8A Pending CN115825025A (en) | 2022-10-12 | 2022-10-12 | Application of chiral carbon quantum dots |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115825025A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116509775A (en) * | 2023-07-05 | 2023-08-01 | 科妍生物工程研究(天津)有限公司 | Rosewood seed skin care lotion and preparation method and application thereof |
-
2022
- 2022-10-12 CN CN202211248927.8A patent/CN115825025A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116509775A (en) * | 2023-07-05 | 2023-08-01 | 科妍生物工程研究(天津)有限公司 | Rosewood seed skin care lotion and preparation method and application thereof |
| CN116509775B (en) * | 2023-07-05 | 2023-10-03 | 科妍生物工程研究(天津)有限公司 | Rosewood seed skin care lotion and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | One-step hydrothermal synthesis of photoluminescent carbon nanodots with selective antibacterial activity against Porphyromonas gingivalis | |
| Huang et al. | Facile and green synthesis of photoluminescent carbon nanoparticles for cellular imaging | |
| Nagvenkar et al. | A one-step sonochemical synthesis of stable ZnO–PVA nanocolloid as a potential biocidal agent | |
| Khan et al. | Targeted highly sensitive detection of multi-drug resistant salmonella DT104 using gold nanoparticles | |
| CN108300464B (en) | Preparation method of antibacterial N-doped carbon quantum dot, product and application thereof | |
| Dong et al. | Preparation of magnetically separable N-halamine nanocomposites for the improved antibacterial application | |
| CN1243059C (en) | Manufacture process of nano fibroin partical | |
| CN108774519B (en) | Ascorbic acid detection method | |
| Shivananda et al. | RETRACTED ARTICLE: Biosynthesis of colloidal silver nanoparticles: Their characterization and potential antibacterial activity | |
| CN115825025A (en) | Application of chiral carbon quantum dots | |
| CN106997799A (en) | A kind of preparation method and its SERS application of high-performance gold shell magnetic microballoon | |
| Ramteke et al. | Synthesis and broad spectrum antibacterial activity of magnetite ferrofluid | |
| Hsu et al. | Is the effect of surface modifying molecules on antibacterial activity universal for a given material? | |
| CN108872171A (en) | A kind of application of fluorescent carbon point and preparation method thereof and detection tetracycline | |
| CN112390282A (en) | Hydrophilic zinc oxide quantum dot and preparation method and application thereof | |
| CN111154485B (en) | Preparation of sulfur-nitrogen double-doped carbon quantum dot and application of sulfur-nitrogen double-doped carbon quantum dot in tetracycline detection | |
| CN108865132B (en) | Fluorescent carbon quantum dot and preparation method and application thereof | |
| Yan et al. | Synthesis and characterization of fluorescent chitosan-ZnO hybrid nanospheres | |
| CN109122678A (en) | A kind of nano-silver loaded is in the nano-ag composite and its preparation method and purposes of amphipathic biological carbon material | |
| CN103332752B (en) | Method for preparing monodispersed alpha-Fe2O3 nanoparticles | |
| Parvin et al. | Polyelectrolyte carbon quantum-dots: new player as a noninvasive imaging probe in Drosophila | |
| Wang et al. | Synergistically enhanced photothermal transition of a polyoxometalate/peptide assembly improved the antibiofilm and antibacterial activities | |
| CN112408483B (en) | Benzoic acid functionalized modified non-stoichiometric molybdenum oxide nanocrystal and preparation method and application thereof | |
| CN111514308B (en) | PH-induced charge-inversion antibacterial gold nanorod and preparation method and application thereof | |
| CN110496232B (en) | Magnetic-gold dimer multifunctional nano probe and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |