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Lei Geng

Publications and source records attributed to Lei Geng.

12 recordsLinked to original sources
PubMed2005

Probing strong adsorption of solute onto C18-silica gel by fluorescence correlation imaging and single-molecule spectroscopy under RPLC conditions.

Understanding molecular adsorption at a chromatographic interface is of great interest for addressing the tailing problem in chemical separations. Single-molecule spectroscopy and confocal fluorescence correlation imaging are used to study the adsorption sites of C(18) silica beads under RPLC chromatographic conditions. The experiments show that cationic molecule rhodamine 6G laterally diffuses through the chromatographic interface of a C(18) hydrocarbon monolayer and acetonitrile with occasional reversible strong adsorptions. Fluorescence correlation imaging extracts the rare strong adsorption events from large data sets, revealing that the strong adsorption sites are randomly distributed throughout the silica beads. Virtually every imaging pixel of silica beads adsorbs molecules. Single-molecule spectroscopy of the 584 strong adsorption events observed indicates that the strong adsorptions persist on the time scales from several milliseconds to seconds, having an average desorption time of 61 ms. The strong adsorption events are rare, comprising 0.3% of the total observation time. The sizes of strong adsorption sites are within the optical resolution of confocal imaging.

Acetonitriles↗
PubMed2005

Correlation coefficient mapping in fluorescence spectroscopy: tissue classification for cancer detection.

Correlation coefficient mapping has been applied to intrinsic fluorescence spectra of colonic tissue for the purpose of cancer diagnosis. Fluorescence emission spectra were collected of 57 colonic tissue sites in a range of 4 physiological conditions: normal (29), hyperplastic (2), adenomatous (5), and cancerous tissues (21). The sample-sample correlation was used to examine the ability of correlation coefficient mapping to determine tissue disease state. The correlation coefficient map indicates two main categories of samples. These categories were found to relate to disease states of the tissue. Sensitivity, selectivity, predictive value positive, and predictive value negative for differentiation between normal tissue and all other categories were all above 92%. This was found to be similar to, or higher than, tissue classification using existing methods of data reduction. Wavelength-wavelength correlation among the samples highlights areas of importance for tissue classification. The two-dimensional correlation map reveals absorption by NADH and hemoglobin in the samples as negative correlation, an effect not obvious from the one-dimensional fluorescence spectra alone. The integrity of tissue was examined in a time series of spectra of a single tissue sample taken after tissue resection. The wavelength-wavelength correlation coefficient map shows the areas of significance for each fluorophore and their relation to each other. NADH displays negative correlation to collagen and FAD, from the absorption of emission or fluorescence resonance energy transfer. The wavelength-wavelength correlation map for the decay set also clearly shows that there are only three fluorophores of importance in the samples, by the well-defined pattern of the map. The sample-sample correlation coefficient map reveals the changes over time and their impact on tissue classification. Correlation coefficient mapping proves to be an effective method for sample classification and cancer detection.

Adenomatous Polyposis Coli↗
PubMed2005

Direct observation of frits and dynamic air bubble formation in capillary electrochromatography using confocal fluorescence microscopy.

Confocal fluorescence microscopy has been used to study the capillary electrochromatography (CEC) frits and dynamic air bubble formation under real chromatographic conditions. Confocal fluorescence microscopy provides a nondestructive way to view the three-dimensional structure of the frits with high spatial resolution. Frits prepared with four different procedures were studied: (1) sintering bare silica beads with sodium silicate; (2) sintering bare silica beads wetted with water; (3) sintering C18 beads wetted with water; and (4) sintering C18 beads wetted with water and then surfaced-recovered with C18. Frits prepared with sintering silicate-wetted beads have a high degree of heterogeneity, while the other three types of frits have similar, more homogeneous packing structures. Confocal fluorescence microscopy also provides sufficient temporal resolution for in situ observation of the dynamic processes in air bubble formation. In this study, air bubble formation is imaged during the reorganization process of the packing bed and is shown to occur close to the border between the packing bed and the outlet frit. Confocal fluorescence microscopy opens a new avenue in studying dynamic processes in situ in CEC separations.

Chromatography, Micellar Electrokinetic Capillary↗
PubMed2005

Statistical and generalized two-dimensional correlation spectroscopy of multiple ionization States. Fluorescence of neurotransmitter serotonin.

Fluorescence spectra of neurotransmitter serotonin are analyzed with generalized and statistical two-dimensional correlation spectroscopy. A comparison is provided for these two emerging data analysis techniques. Both methods reveal correlations between spectral variables and demonstrate enhanced sensitivity in detecting the dynamic spectral changes over conventional one-dimensional spectroscopy. Both statistical and generalized 2D correlation analysis emphasize simultaneous spectral changes in response to external perturbations. Generalized 2D correlation spectroscopy further reveals the difference in rates of these dynamic changes. Using 2D correlation analysis, a third ionization species of serotonin is identified using pH and excitation wavelength perturbation. This species is a doubly deprotonated serotonin with very low fluorescence quantum yield, confirmed by using a laser excitation at longer wavelength and at higher pH. Taking advantage of the spectral differences between excitation of serotonin and tryptophan, as low as 3.8 nM serotonin can be detected in the presence of 20 microM tryptophan, with long-wavelength excitation. This represents the sensitive detection of serotonin in 5000-fold excess of tryptophan.

Fluorescence↗
PubMed2005

Investigation of the stability of polyelectrolyte multilayer coatings in open-tubular capillary electrochromatography using laser scanning confocal microscopy.

A simple polyelectrolyte multilayer (PEM) coating procedure was used for the development of stable modified capillaries. PEM coatings were constructed in fused-silica capillaries using alternating rinses of cationic and anionic polyelectrolytes. The multilayer coatings investigated in this study consisted of two and twenty layer pairs, or bilayers. A bilayer is one layer of a cationic polymer and one layer of an anionic polymer. Poly(diallyldimethylammonium chloride) was used as the cationic polymer, and the polymeric surfactant poly(sodium N-undecanoyl-L-leucylvalinate) was used as the anionic polymer. Previous studies for both chiral and achiral separations have shown that PEM-coated capillaries have excellent reproducibilities, remarkable endurance, and strong stabilities against extreme pH values when used in open-tubular capillary electrochromatography (OT-CEC). In this study, the stability of the coatings was further investigated after exposure to 0.1 M and 1.0 M NaOH. Structural changes of these coatings were monitored using laser scanning confocal microscopy (LSCM) after flushing the capillaries with NaOH. This technique allowed observation of the degradation of the coatings. Observations are discussed in terms of separations using OT-CEC. Electropherograms obtained from the chiral separation of 1,1'-binaphthyl-2,2'-dihydrogenphosphate in OT-CEC showed a decrease in selectivity and an increase in electroosmotic mobility after long exposure to NaOH. The ability to recover the capillaries by exposure to NaOH was also demonstrated. Measurements of electroosmotic mobility and selectivity showed that 2-bilayer and 20-bilayer PEM coatings could be completely removed from the capillary surface after approximately 3.5 and 9.5 h, respectively, of continuous exposure to 1 M NaOH.

Chromatography, Micellar Electrokinetic Capillary↗
PubMed2005

[Study on differential display genes of tolerant-damage of MRC-5 induced by formaldehyde of low dose].

OBJECTIVE: To observe the differentially expressed genes of the human embryo lung fibroblast (MRC-5) induced by formaldehyde (FA) of low dose using fluoro DD-PCR. METHODS: The dose-effect relation of FA toxicity to MRC-5 was acquired, No observed damage effect or proliferation concentration was used as low dose, and obvious damage concentration was used as high dose. MRC-5 was treated with low and high dose, and treated with high dose after pretreated with low dose for some time. Then Fluoro differential display polymerase chain reaction (Fluoro DD-PCR) was used to search differentially expressed genes of the differently treated groups of FA. 61 differential display straps were acquired and 11 of them were reamplified, cloned, sequenced and blasted. RESULTS: According to the dose-effect relation of FA toxicity to MRC-5, 100 micromol/L was choosed as low dose and 10 mmol/L was choosed as high dose. Samples of differently treated groups were amplified by means of fluro-DD-PCR, 61 differentially expressed straps were acquired. 11 differential display straps have been cloned, sequenced and blasted. Two of them were known genes: one was highly homologous to nuclear factor of activated T-cells 5 (NFAT5) and the other was highly homologous to tetratricopeptide repeat domain 3(TPRD-3). Nine of them were new genes. CONCLUSION: It seemed that FA of low dose could promote MRC-5 proliferation and find 61 differentially expressed genes of differently treated groups, and the result of clone could provide scientific base for mechanism of FA toxicity research.

Cell Line↗
PubMed2005

[Research on differential display genes of tolerant-damage induced by trichloroethylene].

OBJECTIVE: To observe the differentially expressed genes of the human normal liver cell (L-02) induced by low concentration of trichloroethylene (TCE). METHODS: The dose-effect relation of TCE toxicity was analyzed by means of MTT. 5 micromol/L was chosen as low concentration while 40 micromol/L as high concentration. Then L-02 were treated with low concentration, high concentration, pretreated with low concentration then attacked with high concentration of TCE respectively. Fluorescence differential display polymerase chain reaction (Fluoro DD-PCR) was used to search differentially expressed genes of the different treatment groups of TCE. Results 51 differential expressed strap were found, 11 differential straps have been cloned and sequenced. 9 of them were known gene, while 2 of them were new genes. CONCLUSION: The differential straps would be identified to provide scientific base for research deeply the mechanism of low concentration TCE inducing adaptive response of L-02.

Cells, Cultured↗
PubMed2002

Effect of polymerization on the subdomain 3/4 loop of yeast actin.

The Holmes F-actin model predicts a polymerization-dependent conformation change of a subdomain 3/4 loop with a hydrophobic tip (residues 266-269), allowing interaction with a hydrophobic surface on the opposing strand of the filament producing filament stabilization. We introduced cysteines in place of Val(266), Leu(267), and Leu(269) in yeast actin to allow attachment of pyrene maleimide. Pyrene at each of these positions produced differing fluorescence spectra in G-actin. Polymerization decreased the fluorescence for the 266 and 267 probes and increased that for the 269 probe. The direction of the fluorescence change was mirrored with a smaller and less hydrophobic probe, acrylodan, when attached to 266 or 269. Following polymerization, increased acrylamide quenching was observed for pyrene at 266 or 267 but not 269. The 267 probe was the least accessible of the three in G- and F-actin. F-actin quenching was biphasic for the 265, 266, and 269 but not 267 probes, suggesting that in F-actin, the pyrene samples multiple environments. Finally, in F-actin the probe at 266 interacts with one at Cys(374) on a monomer in the opposing strand, producing a pyrene excimer band. These results indicate a polymerization-dependent movement of the subdomain 3/4 loop partially consistent with Holmes' model.

Acrylamides↗
PubMed2002

In situ time-resolved fluorescence spectroscopy in the frequency domain in capillary electrochromatography.

In situ time-resolved fluorescence spectroscopy for capillary electrochromatography (CEC) is described in the frequency domain. Fluorescence decay of the solute molecules is collected directly in the packed stationary phase of the CEC capillary. The fluorescence lifetime profile of the solute molecules reveals the microenvironments they experience in the C18 chromatographic interface. A quartz flow cell and experimental optimization of the signal-to-noise ratio are described that enable the collection of high-quality decay data and subsequent calculation of fluorescence lifetime profiles of the solute molecules. The distribution of pyrene (PY), 1-pyrenemethanol (PY-MeOH), and 1-pyrenebutanol (PY-BuOH) into the C18 stationary phase and the solute-C18 phase interactions are probed, under separation conditions for CEC. All three molecules display a Gaussian distribution of lifetimes, consistent with an ensemble of heterogeneous microenvironments in the C18 stationary phase. The least polar molecule PY diffuses deeply into and interacts extensively with the C18 phase, experiencing high hydrophobicity and significant heterogeneity of microenvironments. The retention order of PY-MeOH, PY-BuOH, and PY in CEC is determined by their interactions with the stationary phase, revealed by their fluorescence lifetime distributions.

Journal Article↗
PubMed2002

Imaging solute distribution in capillary electrochromatography with laser scanning confocal microscopy.

A method for the direct observation of solute molecules interacting with a C18 stationary phase under real separation conditions in capillary electrochromatography (CEC) is investigated. The experiments were performed in a capillary electrochromatographic mode; however, the method and findings are useful both in CEC and revered-phase liquid chromatography. The distribution of solute molecules in the packed capillary is directly imaged with laser scanning confocal fluorescence microscopy. Conventional imaging techniques produce images where the C18 silica beads cannot be distinctively identified as a result of the deep depth of field. The optical sectioning capability of confocal imaging overcomes this problem to afford clearly defined images of the stationary-phase packing and the surrounding mobile phase. Fluorescein molecules are preferentially distributed in the mobile phase under reversed-phase chromatographic conditions. Nile Red and rhodamine 6G molecules prefer the environments of the porous C18 beads. Intensity distributions over time for areas within the stationary-phase beads differ from distributions of areas outside the beads in the mobile phase. Images taken at different depths into the capillary probe the internal structure of the C18 beads. While the internal structures of most beads are porous, confocal images show a small fraction (2%) of the silica beads have porous shells and nonporous cores. The capability of imaging the stationary phase distinctively from the mobile phase opens the possibilities of studying the quality of stationary phase, the structure of the column packing, and the mechanisms of separation.

Journal Article↗