The HUMACTBP2 locus was investigated to collect population genetic data in the Korean population and to evaluate the applicability for the forensic field. An Automatic fluorescent-based sequencer (377 automatic DNA sequencer, ABI) was used to detect amplified fragments of the HUMACTBP2 locus electrophoresed on 4% denaturing polyacrylamide sequencing gels. ACTBP2 allelic ladder consisting of different sizes of 18 alleles was constructed and employed as an internal size standard in combination with a GS-350 size standard for precision of allele-band sizing. By utilizing different fluorescent dyes, both the allelic ladders and samples were able to be analyzed in the same lane by 99% orecision of allele-band sizing. Among the Korean population (n=224), 26 alleles in the range of 239-313 bp are determined. allele No. 6 is found 45 times (0.100) which is mostly frequent, and the rest of allele is distributed with their relative frequency of 0.002-0.100. The comparison between observed and expected numbers of homozygous and heterozygous individuals confirms that ACTBP2 locus is in the state of Hardy-Weinberg equilibrium among the Korean population. The heterozygosity is 0.9389+/-0.0034(93.89%), and the power of discrimination(PD) and power of exclusion(PEX) are calculated to be 0.991(99.1%)and 0.890(89.0%), respectively, showing the high informativeness for individual identification. Thus, these results mean that the HUMACTBP2 locus can effectively be used for the forensic application.
Alleles ; DNA ; Fluorescent Dyes ; Gels ; Genetic Variation*

AIMTo evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application.

METHODSSemen samples from 87 unselected infertile patients were used to perform the following assays: (i) detection of active caspase-3 (n=17); (ii) integrity of the mitochondrial membrane potential (MMP) (n=17); (iii) externalization of phosphatidylserine (EPS) (n=16); and (iv) detection of intact acrosomes via CD46 (n=37). After the assays, 4% paraformaldehyde was added to all aliquots. The fluorescence intensity of each sample was evaluated by flow cytometry on days 0, 3, 7, 10 and 14.

RESULTSDifferences of up to +/-5% positive spermatozoa from the value measured at day 0 were estimated as acceptable deviation. The Caspase-3 FLICA showed mean differences<5% at day 3, 7 and 10. At day 14 the mean difference was 7.6%. In contrast, the disrupted MMP and the EPS detection showed differences>5% at day 3. The CD46-FITC labeling displayed absolute differences<5% CD46-positive spermatozoa at days 3, 7, 10 and 14.

CONCLUSIONAlthough immediate analysis of the fluorescence signals is recommended, it is possible to evaluate caspase-3 activation up to 10 days and CD46 up to 14 days after staining of sperm. The FACS evaluation of MMP and EPS detection should be conducted on the same day.

In vivo fluorescence molecular imaging plays a more and more important role in the observation of diseases, drug research and biology research because of its low cost, simplicity and no ionizing radiation to biological tissue. Herein, the most important parts of the optical fluorescence molecular imaging and their advances are summarized, including fluorescent molecular probes, imaging systems and reconstruction algorithms. The application and development trend of this technology are also introduced in this paper.
Algorithms ; Fluorescent Dyes ; Molecular Imaging ; methods

The convective polymerase chain reaction (CCPCR) uses the principle of thermal convection to allow the reagent to flow in the test tube and achieve the purpose of amplification by the temperature difference between the upper and lower portions of the test tube. In order to detect the amplification effect in real time, we added a fluorophore to the reagent system to reflect the amplification in real time through the intensity of fluorescence. The experimental results show that the fluorescence curve conforms to the S-type trend of the amplification curve, but there is a certain jitter condition due to the instability of the thermal convection, which is not conducive to the calculation of the cycle threshold (CT value). In order to solve this problem, this paper uses the dynamic method, using the double S-type function model to fit the curve, so that the fluorescence curve is smooth and the initial concentration of the nucleic acid can be deduced better to achieve the quantitative purpose based on the curve. At the same time, the PSO+ algorithm is used to solve the double s-type function parameters, that is, particle swarm optimization (PSO) algorithm combined with Levenberg-Marquardt, Newton-CG and other algorithms for curve fitting. The proposed method effectively overcoms PSO randomness and the shortcoming of traditional algorithms such as Levenberg-Marquardt and Newton-CG which are easy to fall into the local optimal solution. The of the data fitting result can reach 0.999 8. This study is of guiding significance for the future quantitative detection of real-time fluorescent heat convection amplification.
Algorithms ; Fluorescence ; Fluorescent Dyes ; Polymerase Chain Reaction

This paper reviews the research progress on live-cell super-resolution fluorescence microscopy, discusses the current research status and hotspots in this field, and summarizes the technological application of super-resolution fluorescence microscopy for live-cell imaging. To date, this field has gained progress in numerous aspects. Specifically, the structured illumination microscopy, stimulated emission depletion microscopy, and the recently introduced minimal photon fluxes microscopy are the current research hotspots. According to the current progress in this field, future development trend is likely to be largely driven by artificial intelligence as well as advances in fluorescent probes and relevant labelling methods.
Artificial Intelligence ; Microscopy, Fluorescence ; Fluorescent Dyes ; Technology

Flow cytometry is a multi-parameter, rapid and efficient method for qualitative analysis and quantitative determination of various fluorescently labeled particles in liquid flow. Flow cytometry has been applied in multiple disciplines such as immunology, virology, molecular biology, cancer biology and infectious disease monitoring. However, the application of flow cytometry in plant research is hampered due to the special composition and structure of plant tissues and cells, such as cell walls and secondary metabolites. In this paper, the development, composition and classification of flow cytometry were introduced. Subsequently, the application, research progress and application limitations of flow cytometry in plant field were discussed. At last, the development trend of flow cytometry in plant research was prospected, which provides new perspectives for broadening the potential application scope of plant flow cytometry.
Flow Cytometry/methods* ; Plants ; Fluorescent Dyes

It is well known that the apposition of bone at implant surface would be influenced by the microstructure of titanium implants. The purpose of this study was to compare bone healing around the screw-shaped titanium implant with three different surface topographies in the canine mandibles by histological and biomechanical evaluation. All mandibular premolars of six mongrel dogs were extracted and implants were placed one month later. The pure titanium implants had different surface topographies: smooth and machined (Steri-Oss(R): Group I); acid-etched (Osseotite(R): Group II); sandblasted and acid-etched (ITI(R), SLA: Group III) surface. The fluorescent dyes were injected on the 2nd (calcein), 4th (oxytetracycline HCl) and 12th (alizarin red) weeks of healing. Dogs were sacrificed at 4 and 12 weeks after implantation. The decalcified and undecalcified specimens were prepared for histological and histometrical evaluation of implant-bone contact. Some specimens at 12 weeks after implantation were used for removal torque testing. Histologically, direct bone apposition to implant surface was found in all of the treated groups. More mature and dense bone was observed at the implant-bone interface at 12 weeks than that at 4 weeks after implantation. Under the fluorescent microscope, thick regular green fluorescent lines which mean early bone apposition were observed at the implant-bone interface in Group III, while yellow and red fluorescent areas were found at the implant-bone interface in Group I and II. The average implant-bone contact ratios at 4 weeks of healing were 54.3% in Group I, 57.7% in Group II and 66.2% in Group III. In Group I, implant-bone contact ratio was significantly lower than Group II and III(p<0.05). The average implant-to-bone contact ratios at 12 weeks after implantation were 64.3% in Group I, 66.7% in Group II and 71.2% in Group III. There was no significant difference among the three groups. In Group I and II, the implant-bone contact ratio at 12 weeks increased significantly in comparison to ratio at 4 weeks(p<0.05). The removal torque values at 12 weeks after implantation were 90.9 Ncm in Group I, 81.6 Ncm in Group II and 77.1 Ncm in Group III, which were significantly different(p<0.05). These results suggest that bone healing begin earlier and be better around the surface-treated implants compared to the smooth surface implants. The sandblasted and acid-etched implants showed the most favorable bone response among the three groups during the early healing stage and could reduce the waiting period prior to implant loading.
Animals ; Bicuspid ; Dogs ; Fluorescent Dyes ; Mandible ; Titanium* ; Torque

Over the past 40 years, flow cytometry has emerged as a leading, application-rich technology that supports high-resolution characterization of individual cells which function in complex cellular networks such as the immune system. This brief overview highlights advances in multiparameter flow cytometric technologies and reagent applications for characterization and functional analysis of cells modulating the immune network. These advances significantly support high-throughput and high-content analyses and enable an integrated understanding of the cellular and molecular interactions that underlie complex biological systems.
Antibodies ; Flow Cytometry ; Fluorescent Dyes ; Immune System ; Immunophenotyping

OBJECTIVETo prepare a pH fluorescence probe based on styrylcyanine dyes for live cell imaging.

METHODSThe Probe 1 was prepared by reaction of 4-pyridinecarboxaldehyde with 1,1,2-trimethylbenz[e]indole. The influence of pH on the fluorescent properties was examined, and the cell viability was examined using cell counting kit-8. The Probe 1 was used as a pH fluorescence probe in living cell.

RESULTSProbe 1 emitted green fluorescence under neutral and basic conditions but orange fluorescence under acid condition. Probe 1 selectively stained the cytoplasmic regions of living cells without significantly affecting the cell viability.

CONCLUSIONThe pH-sensitive fluorescent probe prepared based on styrylcyanine possesses good ability of cell membrane permeation for live cell fluorescent imaging.

As a new type of carbon nanomaterials, fluorescent carbon dots (fluorescent CDs) have many advantages when compared with the traditional fluorescent probes. They are photoluminescence stable and resistance to photo bleaching. Moreover, they are excellent in biocompatibility, low-toxic and easy to modify. All these above make them a promising optical image material as a probe in optical image. This article reviews structure, the common carbon sources, the preparation methods, and the light-emitting principles of the carbon dots. We also introduce the research progress of fluorescent carbon dots in biomedicine, and the problems need to be resolved in the study of fluorescent CDs.
Carbon ; chemistry ; Fluorescent Dyes ; chemistry ; Nanostructures ; chemistry ; Quantum Dots ; chemistry