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两种体外细胞毒性检测方法的比较研究 总被引:8,自引:0,他引:8
目的比较两种常用的细胞毒性检测方法在医疗器械生物学评价中的相关性。方法分别采用MTT比色法和细胞增殖度法,在37℃条件下,将五种医疗器械/生物材料的浸提液分别与小鼠成纤维细胞(L-929)接触2天和2,4,7天,比较材料对细胞的毒性影响。结果5种不同的材料浸提液分别表现出不同程度的细胞毒性反应(0~2级)。将MTT比色法与细胞增殖度法(2天)的实验数据进行相关性分析,显示两者之间具有良好的相关性(R=0.977)。结论MTT比色法由于其检测所需的细胞量相对较少,试验步骤相对简便、检测周期短,因此具有一定的优越性,是个值得推荐的细胞毒性检测方法。 相似文献
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越来越多研究证明线索细胞(clue cell)是细菌性阴道病的重要致病因子,而细菌性阴道病(bacterial vaginosis BV)是妇产科最常见的疾病之一,感染率在15%~50%,且易复发[1,2].在BV的诊断中线索细胞检查是一项重要的指标.我们采用妇科细胞学涂片快速多项染色法(CTB)和盐水湿片法检查白带中的线索细胞,并且进行比较,了解二种方法的特异性和敏感性. 相似文献
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主要介绍了一类基于荧光检测的新型细胞传感器,这类传感器利用免疫细胞表面分子特异性识别、结合抗原的特性和生物(或化学)发光技术,通过检测荧光信号在数分钟内达到检测病原体或其他抗原的目的.这类传感器的发光原理主要是利用钙离子敏感型化学荧光探针发光,如Fluo-4等,或钙离子敏感型发光蛋白发光,如水母发光蛋白、绿色荧光蛋白等.现在已经应用的主要是B细胞传感器和肥大细胞传感器.这类传感器具有灵敏度高、检测准确、反应速度快的优点.同时又存在交叉反应、细胞不易保存等不足之处.这类传感器在疾病诊断、环境监测、生物战剂检测等领域具有较大的应用前景. 相似文献
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目的通过牛肾细胞在两种不同载体中培养效果的比较,为牛肾细胞在细胞工厂中规模化生产提供真实的、有力的支持。方法不同代次牛肾细胞在两种载体中经过相同培养条件进行培养。结果实验中原代牛肾细胞在细胞工厂接种密度为5.5×104/cm2左右,在15 L转瓶接种密度为9.0×104/cm2左右。一代牛肾细胞在细胞工厂接种密度为6.5×104/cm2左右,在15 L转瓶接种密度为10×104/cm2左右。二代牛肾细胞在细胞工厂接种密度为7.0×104/cm2左右,在15 L转瓶接种密度为14×104/cm2左右。两种载体中牛肾细胞生长状况均能达到培养要求。结论细胞工厂能在有限的空间内利用最大限度的培养表面培养牛肾细胞,不仅节约了传代前的细胞用量,而且提高了培养后的细胞产量。 相似文献
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细胞凋亡检测技术的进展 总被引:7,自引:0,他引:7
细胞凋亡是多细胞生物体内一个重要的生命现象,它已成为当前科学和医学领域的研究热点。随着对细胞凋亡的研究不断扩展和深化,许多新的指标逐渐被采纳;同时伴随着一些先进仪器的应用,原有一些指标的灵敏性和准确性也得到了提高,这些都将细胞凋亡的检测水平提升到一个新的高度。 相似文献
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Comparison of 5 microplate colorimetric assays forin vitro cytotoxicity testing and cell proliferation assays 总被引:1,自引:0,他引:1
This paper describes a critical comparative evaluation of 5 miniaturised colorimetric assays applicable to cytotoxicity testing of anti-tumour drugs (and other toxins)in vitro. Each assay shows a different linear range for optical density versus cell number, a different sensitivity to change in cell number and a different minimum detectable cell number; the values of these parameters vary with experimental conditions and with cell line used. All the methods gave good correlation with viable cell number (determined by colony forming efficiency) in toxicity assays after 3 or 4 days of treatment, but they underestimated cell death after 2 days. Toxicity levels for individual chemicals (in a standard 6-day assay) are similar for the different assays, irrespective of the mechanism of action of the chemical being tested. Two of the more recently developed assays (APNaOH and SRB) were found to be very sensitive under the conditions examined. 相似文献
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Parida MM Pandya G Bhargava R Bhattacharya R Jana AM 《Indian journal of experimental biology》1999,37(12):1223-1226
Efficacy of two colorimetric assays, viz. MTT (3-4,5-dimethylthiazol-2-(yl-2,5-diphenyl tetrazolium bromide) and neutral red (NR) assays, performed by integrating them to micro culture virus titration (MCVT), was compared with the conventional MCVT method in terms of percentages of infectivity and 50% infectivity end points by employing Polio virus type-3 and Dengue virus type 4 as the candidate viruses. The results suggested that MTT assay has an edge over NR assay as well as conventional MCVT method. For the first time, NR assay has been successfully employed for the determination of virus infectivity titre. 相似文献
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Henriksson E Kjellén E Wahlberg P Wennerberg J Kjellström JH 《In vitro cellular & developmental biology. Animal》2006,42(10):320-323
Summary The aim of this study was to evaluate some bioassays that are different in principle: cell counting, colony forming assay,
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), sulforhodamine B (SRB), crystal violet, and alamarBlue,
with respect to their ability to measure cisplatin-induced cell death of in vitro-cultivated squamous cell carcinoma of the
head and neck (SCCHN). Cisplatin was applied in concentrations of 1.0, 5.0, 10.0, 50.0, and 100 μM. The cells were incubated for 1 h, and the cell survival was measured 5 d after treatment. We found the colorimetric assays
and cell counting to be comparable. The colony forming assay indicated a higher degree of cell kill compared with the other
techniques. Measurement of cell survival after treatment with cisplatin can be done by use of any of the above tested assays.
However, the majority of SCCHN cell lines available do not form colonies easily, or at all. Therefore, comparing the chemosensitivity
between such cell lines is limited to alternative assays. In this respect, any of the tested colorimetric assays can be used.
However, they seem to underestimate cell kill. Cell counting is also an alternative. This technique, however, is time consuming
and operator dependent, as in the case of manual counting, or relatively expensive when counting is performed electronically,
compared with the colorimetric assays. 相似文献
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Ekaterina Kozaeva Viviënne Mol Pablo I. Nikel Alex Toftgaard Nielsen 《Microbial biotechnology》2022,15(9):2426-2438
Randomized strain and pathway engineering are critical to improving microbial cell factory performance, calling for the development of high-throughput screening and selection systems. To facilitate this effort, we have developed two 96-well plate format colorimetric assays for reliable quantification of various ketones and aldehydes from culture supernatants, based on either a vanillin-acetone reaction or the 2,4-dinitrophenylhydrazine (2,4-DNPH) reagent. The vanillin-acetone assay enabled accurate and selective measurement of acetone titers up to 2 g l−1 in a minimal culture medium. The 2,4-DNPH-based assay can be used for a wide range of aldehydes and ketones, shown here through the optimization of conditions for 15 different compounds. Both assays were implemented to improve acetone production from different substrates by an engineered Escherichia coli strain. The fast and user-friendly colorimetric assays proposed here open the potential for iterative rounds of (automated) strain and pathway engineering and screening, facilitating the efforts towards further boosting production titers of industrially relevant ketones and aldehydes. 相似文献
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Norma L. Fowler 《Oecologia》1984,62(3):424-428
Summary Individuals of two perennial bunch grasses, Bouteloua rigidiseta and Aristida longiseta, were transplanted into otherwise undisturbed natural short-grass vegetation in which these species are dominants. Significant differences in survivorship and growth rate were found among quadrats for both species. The relative favorability of quadrats for Bouteloua, but not for Aristida, changed during the course of the two year study. For neither species was a consistent relationship between pre-existing density and transplant performance found. However, one patch type was identified in which Bouteloua transplants grow well, although this species is not naturally present there, apparently because it cannot establish. Microsite differences, created by different soil in transplanted peat pots, were also found to be important. Bouteloua had a higher survivorship the first year, Aristida the second, which suggests that Aristida is the more tolerant of drier years. 相似文献
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The effects of various bioactive sphingolipids (sphingosine 1-phosphate, sphingosine 1-phosphocholine, ceramide 1-phosphate, ceramide beta-glucoside and beta-lactoside, and gangliosides) on cell proliferation and apoptosis are reviewed. It is concluded that the balance between the bioeffector sphingolipids determines their overall effect on cell. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 3; see also http://www.maik.ru. 相似文献
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Two-hybrid systems have become favored tools for detection and analysis of protein interactions because of their low cost and ease of use compared to biochemical or biophysical interaction technologies. It is possible to augment the utility of two-hybrid systems and derivative systems such as dual-bait two-hybrid systems by adapting strategies that speed the analysis of the relative strength of a series of protein-protein associations. This report describes two simple techniques that employ either a flatbed scanner or a plate reader to quantitate the activity of colorimetric reporters such as LacZ or GusA commonly used in two-hybrid approaches. 相似文献
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In this paper, we describe two simple, label-free, homogenous assays using gold nanoparticles (Au NPs)-one to detect coagulation-related proteins and the other to screen inhibition reactions. The first nanosensor functions on the basis of the fact that thrombin catalyzes fibrinogen to form long-chain fibrins, which then induce aggregation of Au NPs. We applied this sensor to study the interactions of thrombin, inhibitors, cofactors, and antidotes. We further used thrombin-conjugated Au NPs (Thr-Au NPs) to analyze the levels of fibrinogen in plasma samples via fibrinogen-induced aggregation of Thr-Au NPs. The limit of detection (LOD; S/N=3) of this sensor for fibrinogen in plasma was 10nM. The Thr-Au NP probe provided quantitative results for fibrinogen in plasma samples that correlated (R(2)=0.97) with those obtained using a clinical von Clauss clotting rate assay. In addition, the Thr-Au NP-based sensor could be used to monitor thrombin concentrations in plasma samples under physiological conditions. Compared with conventional assays, these label-free assays offer several advantages, such as rapid and simple readout by the naked eye or by UV-vis absorption spectroscopy. 相似文献