Comparison of Fluorometric and UV Spectrophotometric Findings for DNA Isolated From Formalin-Fixed Paraffin-Embedded Blocks, Fine Needle Aspiration Cytology Smears, and Blood
Introduction: Fine needle aspiration cytology (FNAC) smear may serve as a convenient sample for DNA extraction for molecular pathology in addition to more commonly used formalin-fixed paraffin-embedded (FFPE) sections. DNA quantification done by fluorometer is more accurate than UV vis spectrophotometer regardless of the source. This study was conducted to compare DNA yield and quality from cytology smears, FFPE sections and peripheral blood using both fluorometer and spectrophotometer. Further, introspection was made to check for the adequacy of DNA extracted from cytology smears with respect to DNA extracted from core biopsies.
Method: DNA was extracted from 10 fresh peripheral blood samples, core biopsies and FNAC smears. The DNA was quantified using a fluorimeter and UV vis spectrophotometer in all cases.
Results: Statistically significant difference was seen between the data obtained from UV vis spectrophotometry and flourometry. The quantity of DNA extracted from FNAC smears was higher than that of core biopsy as per fluorometry data (mean DNA of core biopsy = 1.9ng/µl, of FNAC = 3.3ng/µl).
Conclusion: DNA estimation by fluorometry is more accurate and precise than spectrophotometry in FFPE, FNAC and whole blood samples. DNA yield from FNAC slides is comparable to that from core biopsies.
Novel high throughput sequencing – fluorometric approach demonstrates Microcystis blooms across western Lake Erie are promoted by grazing resistance and nutrient enhanced growth
Cyanobacterial harmful algal blooms (CHABs) are a global public health threat. While CHABs are often promoted by nutrients, an important and often overlooked influence on bloom dynamics is zooplankton grazing. In the present study, zooplankton grazing and nutrient enrichment experiments were combined with next generation sequencing and fluorometric analyses to quantify differential grazing and nutrient effects on specific cyanobacterial genera across the western basin of Lake Erie. Grazing by two different sized daphnids, Daphnia magna and Daphnia pulex, was compared to protozooplankton grazing effects assessed via a dilution approach at sites within the Maumee and Sandusky Bays where Planktothrix, Microcystis, Synechococcus, and Dolichospermum were the dominant genera. Daphnid grazing significantly reduced Synechococcus net growth rates at most sites as well as Planktothrix net growth in Sandusky Bay and Dolichospermum in Maumee Bay.
Dilution resulted in significant growth increase of Synechococcus at half of the sites and Planktothrix at most sites evidencing substantial grazing pressure by the protozooplankton community on these genera. In contrast, Microcystis populations were largely unaffected by daphnids and protozooplankton grazing but benefitted from nutrient enrichment more than other CHAB genera. When diatoms were present in moderate abundance, grazing rates by daphnids on diatoms were significantly greater than grazing rates on cyanobacteria. The novel approach used in this study established differences in grazing pressure and nutrient effects on differing taxa and revealed that, while many taxa were grazed by multiple classes of zooplankton (e.g. Planktothrix, Synechococcus, Dolichospermum, diatoms), the lack of grazing pressure on Microcystis coupled with nutrient-enhanced growth in western Lake Erie promotes the occurrence of CHABs of this genus.
Fluorometric determination of ziram using CsPbBr 3 quantum dots
A strategy based on CsPbBr3 quantum dots (QDs) is described for the determination of ziram pesticide. A facile and inert gas-free method was used for the synthesis of CsPbBr3 QDs. The obtained CsPbBr3 QDs displayed turn-off fluorescence behavior for ziram. The fluorescence intensity of the CsPbBr3 QDs (Ex/Em = 365/516 nm) was inversely proportional to the concentration of ziram (0.10 to 50.0 ppm) with a detection limit of 0.086 ppm. Notably, satisfactory recoveries (100 ± 0.25 to 107 ± 5.72%) were obtained in spiked fruit samples, which demonstrated that this method is capable of detecting ziram in real samples.
In addition, the mechanism for the detection of ziram was investigated in detail. According to the results, this mechanism can be tentatively explained by fluorescence quenching originating from the increased surface defects and the structural changes of the CsPbBr3 QDs. The detection ability of this strategy shows promising applicability in food safety.
Spectrophotometric and fluorometric detection of DNA/BSA interaction, antimicrobial, anticancer, antioxidant and catalytic activities of biologically active methoxy substituted pyrimidine-ligand capped copper nanoparticles
- New Schiff base ligand (DPMN) was synthesized from the condensation of 2-hydroxy-5-nitrobenzaldehyde and 2-amino-4,6-dimethoxypyrimidine which was confirmed by spectroscopic and analytical methods. Solid air stable copper nanoparticles (DPMN-CuNPs) were synthesized from its copper chloride salt and it is stabilized by the prepared Schiff base ligand by phase transfer assisted synthesis which is a modified Brust-Schiffrin technique. The formation of ligand stabilized copper nanoparticles was confirmed by UV-Visible and FT-IR spectroscopic techniques. The size, surface morphology and quality of DPMN-CuNPs were analyzed by SEM and TEM techniques. Antioxidant activities of DPMN and DPMN-CuNPs with DPPH, SOD, peroxide and nitrous oxide were analyzed by electronic absorption spectroscopy.
- DNA interaction between DPMN and DPMN-CuNPs with CT-DNA was carried out using electronic absorption, fluorescence, viscometric measurements and cyclic voltammetric techniques. Interaction between BSA and the synthesized compounds analyzed by electronic absorption spectroscopy, Antimicrobial studies confirmed that the synthesized DPMN-CuNPs possess significant biological activities than DPMN. Anticancer results suggest that prepared DPMN-CuNPs have significant anticancer activity against different cancer cell lines and least toxic effect against the normal (NHDF) cell line. Other than the positive response in biological evaluation, our DPMN-CuNPs possess good catalytic activity in methyl orange reduction, methylene blue degradation and nitro phenol reduction.
Imidazole for Fluorometric |
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| 19028-22 | NACALAI TESQUE | 25G | 95.2 EUR |
Imidazole for Fluorometric |
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| 19028-64 | NACALAI TESQUE | 100G | 237.3 EUR |
GST Fluorometric Assay Kit |
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| 55R-1350 | Fitzgerald | 100 assays | 513 EUR |
NADH Fluorometric Assay Kit |
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| K2037-100 | ApexBio | 100 assays | 557 EUR |
Zinc Assay Kit (Fluorometric) |
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| K428-100 | Biovision | each | 627.6 EUR |
HDAC Fluorometric Assay kit |
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| TBS2060 | Tribioscience | 100 tests | 289 EUR |
Urea Assay Kit (Fluorometric) |
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| MET-5180 | Cell Biolabs | 200 assays | 452 EUR |
Starch Assay Kit (Fluorometric) |
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| MET-5026 | Cell Biolabs | 100 assays | 410 EUR |
Malate Assay Kit (Fluorometric) |
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| MET-5120 | Cell Biolabs | 100 assays | 395 EUR |
Lysine Assay Kit (Fluorometric) |
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| MET-5129 | Cell Biolabs | 100 assays | 525 EUR |
Starch Assay Kit (Fluorometric) |
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| MBS169282-100Assays | MyBiosource | 100Assays | 580 EUR |
Starch Assay Kit (Fluorometric) |
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| MBS169282-5x100Assays | MyBiosource | 5x100Assays | 2655 EUR |
Malate Assay Kit (Fluorometric) |
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| MBS169576-100Assays | MyBiosource | 100Assays | 560 EUR |
Malate Assay Kit (Fluorometric) |
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| MBS169576-5x100Assays | MyBiosource | 5x100Assays | 2575 EUR |
Lysine Assay Kit (Fluorometric) |
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| MBS169586-100Assays | MyBiosource | 100Assays | 710 EUR |
Lysine Assay Kit (Fluorometric) |
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| MBS169586-5x100Assays | MyBiosource | 5x100Assays | 3275 EUR |
Glucose Fluorometric Assay Kit |
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| K2221-100 | ApexBio | 100 assays | 571 EUR |
Lactate Fluorometric Assay Kit |
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| K2140-100 | ApexBio | 100 assays | 586 EUR |
Inosine Fluorometric Assay Kit |
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| K712-100 | Biovision | each | 639.6 EUR |
Calcium Assay Kit (Fluorometric) |
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| K409-100 | Biovision | each | 574.8 EUR |
Glycine Assay Kit (Fluorometric) |
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| K589-100 | Biovision | each | 829.2 EUR |
Lactate Assay Kit (Fluorometric) |
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| MET-5013 | Cell Biolabs | 100 assays | 390 EUR |
Choline Assay Kit (Fluorometric) |
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| MET-5042 | Cell Biolabs | 96 assays | 440 EUR |
Guanine Assay Kit (Fluorometric) |
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| MET-5148 | Cell Biolabs | 100 assays | 430 EUR |
Alcohol Assay Kit (Fluorometric) |
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| STA-621 | Cell Biolabs | 100 assays | 450 EUR |
Glucose Assay Kit (Fluorometric) |
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| STA-681 | Cell Biolabs | 500 assays | 465 EUR |
Ethanol Fluorometric Assay kit |
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| TBS2095 | Tribioscience | 100 tests | 259 EUR |
Sucrose Fluorometric Assay Kit |
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| E-BC-F042-96T | Elabscience Biotech | 96T | 350 EUR |
Determination of L-Phenylalanine in Human Plasma Samples with New Fluorometric Method
The measurement of phenylalanine in biological fluids for the diagnosis of phenylketonuria (PKU) in newborns and the monitoring/therapeutic drug monitoring of individuals with PKU are especially important. Owing to the importance of PKU monitoring in clinical medicine, a new fluorometric method was developed for the determination of L-phenylalanine in serum samples. This method is based on the relationship between phenylalanine ammonia-lyase (PAL) and o-phthalaldehyde (OPA). PAL catalyzes the conversion of phenylalanine to ammonia and trans-cinnamic acid. The formed ammonia reacts with OPA in the presence of sodium sulfite, giving a fluorescent product. The presence of sulfide in an alkaline environment prevents OPA from reacting with other amino acids while allowing it to react only with ammonia.
Method characterization and optimization studies, such as the effects of pH, temperature, and interferents, were carried out. The amount of L-phenylalanine in a human serum sample was successfully determined by using the fluorescence emission intensity of the fluorescent product formed as a result of the reaction between OPA and ammonia. The linear range of the method is between 10 μM and 10 mM. The obtained result showed good agreement with the results of the biochemistry analysis laboratory. Recoveries of 95.41% and 73.39% were obtained for phenylalanine and ammonia, respectively. This PAL-OPA-based fluorometric method for phenylalanine is practical, easy to operate, low cost, highly sensitive, and selective and can also be used for the simultaneous determination of ammonia in human serum samples.