蛍光分光光度計・蛍光寿命測定装置
2005年
蛍光分光光度計 SPEX FluoroMaxについて記述のある文献一覧です。
掲載年 | タイトル | 著者 | 掲載誌 | 概要 |
|---|---|---|---|---|
2005 | Green fluorescent protein quantification in whole plants. | Halfhill, Matthew D; Millwood, Reginald J; Stewart, C Neal | journal name-Methods Mol Biol | As future biotechnology applications utilize recombinant proteins as commercial products, nondestructive assays will be necessary to determine protein concentrations accurately within plant tissues. Green fluorescent protein (GFP) has been proposed as a potential marker for the monitoring of transgenic plants and quantifying recombinant protein levels under field conditions. This chapter discusses the utility of using GFP fluorescence as an indicator of protein concentrations and the methods used to quantify GFP fluorescence in whole plant tissues. Furthermore, we discuss the accuracy and effectiveness of the portable General Fluorescence Plant Meter (GFP Meter, Opti-Sciences, Inc.) compared to a laboratory-based spectrofluorometer ( Fluoro-Max2 , Jobin Yvon & Glen Spectra). In whole plants, GFP fluorescence was shown to be variable at each leaf position over time and among different leaves on the same plant. A leaf had its highest GFP fluorescence after emergence, and subsequently, its fluorescence intensity decreased over time. Younger leaves were significantly more fluorescent than older leaves on the same plant. GFP fluorescence intensity was directly correlated with the concentration of soluble protein per unit wet mass and with another genetically linked recombinant protein (Bacillus thuringiensis Bta cry1Ac endotoxin protein). |
2005 | Criticality of pH for accurate fluorometric measurements of dipyridamole levels in biological fluids. | Oshrine, B; Malinin, A; Pokov, A; Dragan, A; Hanley, D; Serebruany, V; Aggrenox Compliance Task Force | journal name-Methods Find Exp Clin Pharmacol | Extended release dipyridamole (DIP) is widely used in clinical practice as an Aggrenox formulation, which is proven to improve outcomes for secondary stroke prevention in patients after acute vascular events. However, presently established fluorometry techniques are not suitable for trace amount determinations, because of the variable background fluorescence. The authors sought to determine whether biological fluid pH is important for the serial measures of DIP levels in the animal experiments and in patients treated with Aggrenox after ischemic stroke. Post-stroke patient (n = 34) and mice (n = 25) samples were tested to determine DIP levels by established techniques with FluoroMax 3 spectrofluorometer. Both the absorption and emission spectra of DIP were affected by modifications in pH. Fluorescence of DIP was found to be maximal at a wavelength of 490 nm (excitation 420 nm) and the spectral pattern was independent of pH. The intensity of fluorescence, however, was drastically lower at low pH (at pH 2.6, fluorescence was 4% of intensity at pH 9.8). Background plasma fluorescence, however, was completely unaffected by changes in pH. Using these fluorometric characteristics, a regression model that facilitates the efficient and sensitive determination of DIP concentration in biological fluids was formulated. Exploiting pH-dependent characteristics of DIP versus serum fluorescence patterns permits a convenient mathematical model to determine DIP concentration. This relatively inexpensive and time-efficient procedure can quantify drug levels in human/animal plasma/serum, thereby directly determining the level of patient adherence to the prescribed drug regimen, be it in the context of clinical trials or compliance with the animal protocol. Copyright 2005 Prous Science. All rights reserved. |
2005 | Autofluorescence characterization of DMBA-TPA induced two-stage carcinogenesis in mouse skin for the early detection of tissue transformation | Anvari, Bahman; Diagaradjane, Parmeswaran; Wong, Michael S.; Yaseen, Mohammed A.; Yu, Jie | abbreviated journal title-Progr. Biomed. Opt. Imaging Proc. SPIE | The use of autofluorescence technique in the characterization of the sequential tissue transformation process in 7,12-dimethylbenz(a)anthracene and 12-O-tetradecanoylphorbol-13-acetate (DMBA & TPA) induced two-stage mouse skin carcinogenesis model in conjunction with a suitable statistical method is being explored. The fluorescence excitation emission matrix (EEM) from experimental group (n=40; DMBA/TPA application), control group (n=6; acetone application) and the blank group (n=6; no application of DMBA/TPA or acetone) were measured every week using Fluoromax3 spectrofluorometer coupled with a waveguide fiber optic bundle (JY Horiba, NJ). The EEM was recorded for 19 excitation wavelengths from 280 to 460 nm at 10 nm intervals and the fluorescence emission was scanned from 300 to 750 nm. During the tissue transformation the epithelial tissues underwent biochemical and structural changes that are manifested in the tissue fluorescence. To correlate the tissue morphology with the observed fluorescence differences in the fluorescence emission, animals were sacrificed and the tissue biopsies were subjected to histopathological evaluation. The fluorescence emission corresponding to different fluorophores was extracted from the EEM, and the spectral data were used in multivariate statistical algorithm for the earliest diagnosis of the onset of tissue transformation. The intrinsic fluorescence from tryptophan, NADH and prophyrins showed distinct differences in the spectral signature during the tissue transformation, due to the altered metabolic activities of the cells. The statistical analysis of the spectral data corresponding to each excitation wavelength showed better classification accuracy at 280, 320, 350 and 405 nm excitations, corresponding to tryptophan, collagen, NADH and porphyrins with the classification accuracy of 74.3, 68.1, 64.6 and 74.7 % respectively. The variations in the spectral signature and the results of the statistical analysis suggest that porphyrins, tryptophan and NADH can be targeted as potential tumor markers in the early detection of the tissue transformation process. 22 Refs. |