A direct heating model to overcome the edge effect in microplates

Lau, Chun Yat; Zahidi, Alifa Afiah Ahmad; Liew, Oi Wah; Ng, Tuck Wah

doi:10.1016/j.jpba.2014.09.021

Cited by 9 publications

(7 citation statements)

References 23 publications

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“…). As was expected from literature , a characteristic temperature distribution is observed during the heating phase with higher temperatures in the outer part than in the inner part of the plate. The heating phase takes about 30–40 min which also agrees well with previous data .…”

Section: Resultssupporting

confidence: 85%

“…Evaporation is assumed to cause these temperature variations within MTPs , and, thereby, also variations of enzyme kinetics . Plate sealing foils are often used to minimize evaporation and, thereby, edge effects . To quantify the temperature offset due to heat losses from evaporation, the MTPs were incubated on a shaken microplate tray in a temperature (36.3 ± 1.0°C) and humidity (45 ± 5%) controlled room.…”

Section: Resultsmentioning

confidence: 99%

“…The high level of parallelization and reduced sample volumes allow the generation of high amounts of quantitative data and, thereby, more rapid bioprocess development . However, a major disadvantage of experimentation with MTP is the susceptibility to spatial effects such as plate‐to‐plate variability, bowl and edge effects .…”

Section: Introductionmentioning

confidence: 99%

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Enzyme activity deviates due to spatial and temporal temperature profiles in commercial microtiter plate readers

Grosch

Sieben

Lattermann

et al. 2016

Biotechnology Journal

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Microtiter plates (MTP) and automatized techniques are increasingly applied in the field of biotechnology. However, the susceptibility of MTPs to edge effects such as thermal gradients can lead to high variation of measured enzyme activities. In an effort to enhance experimental reliability, to quantify, and to minimize instrument-caused deviations in enzyme kinetics between two MTP-readers, we comprehensively quantified temperature distribution in 96-well MTPs. We demonstrated the robust application of the absorbance dye cresol red as easily applicable temperature indicator in cuvettes and MTPs and determined its accuracy to ±0.16°C. We then quantified temperature distributions in 96-well MTPs revealing temperature deviations over single MTP of up to 2.2°C and different patterns in two commercial devices (BioTek Synergy 4 and Synergy Mx). The obtained liquid temperature was shown to be substantially controlled by evaporation. The temperature-induced enzyme activity variation within MTPs amounted to about 20 %. Activity deviations between MTPs and to those in cuvettes were determined to 40 % due to deviations from the set temperature in MTPs. In conclusion, we propose a better control of experimental conditions in MTPs or alternative experimental systems for reliable determination of kinetic parameters for bioprocess development.

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Section: Resultssupporting

confidence: 85%

Section: Resultsmentioning

confidence: 99%

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Enzyme activity deviates due to spatial and temporal temperature profiles in commercial microtiter plate readers

Grosch

Sieben

Lattermann

et al. 2016

Biotechnology Journal

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“…Flat transparency microplates originally developed for enhanced cost effectiveness (34,38,39) may offer a viable alternative to standard deep-well microplates because molding, incorporation of additives, and irradiation treatment can be performed more uniformly during manufacture. These designs have also been demonstrated to be free of cross talk and thermal edge effects (33,3). Further work using other antibody pairs in validated ELISAs will also shed more light on the impact of microplate surface heterogeneities on immunoassay variability.…”

Section: Articlesmentioning

confidence: 97%

“…A major difficulty with immunoassays is assay variation attributed to both microplate-and nonmicroplate-associated factors, and compounded by a complicity of these factors leading to exacerbation of assay variability. Operator competency; microplate, calibrator, and reagent lot differences; microplate edge effects arising from temperature and evaporation rate differentials; and convective forces during assay incubation and washing steps have been reported to contribute to variations in well-to-well and plateto-plate signal read out (1)(2)(3)(4)(5)(6)(7)(8). Microplate variability with row and/or column effects within the microplate as well as systematic differences between microplates has been observed (7,8).…”

mentioning

confidence: 99%

Variability in Microplate Surface Properties and Its Impact on ELISA

Lilyanna

Moriguchi

et al. 2018

The Journal of Applied Laboratory Medicine

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Background: Microplate-based immunoassays are widely used in clinical and research settings to measure a broad range of biomarkers present in complex matrices. Assay variability within and between microplates can give rise to false-negative and false-positive results leading to incorrect conclusions. To date, the contribution of microplates to this variability remains poorly characterized and described. This study provides new insights into variability in immunoassays attributable to surface characteristics of commercial microplates. Methods: Well-to-well assay variation in γ-treated and nontreated 96-well opaque microplates suitable for chemiluminescence assays was determined by use of a validated sandwich ELISA. Microplate surface characteristics were assessed by sessile drop contact angle measurements, scanning electron microscopy, energy dispersive x-ray spectroscopy, and atomic force microscopy. Results: All microplate types tested exhibited vendor-specific assay response profiles; and "rogue" plates with very high intraassay variation and deviant mean assay responses were found. Within-plate, location-dependent bias in assay responses and variability in well contact angle were also observed. We demonstrate substantial differences in well-surface properties with putative effects on protein-coating reproducibility and hence consistency in immunoassay responses. A surface "cleaning" effect on manufacturing residues was attributed to γirradiation, and treated microplates manifest increased polar functionalities, surface roughness, and assay responses. Conclusions: Our data suggest that tighter control of variability in surface roughness, wettability, chemistry, and level of residual contaminants during microplate preparation is warranted to improve consistency of ELISA assay read out. IMPACT STATEMENT In clinical and research settings, microplate-based immunoassays are subject to assay variability within and between microplates that can give rise to false-negative and false-positive results leading to incorrect conclusions. This article clarifies the contribution of microplates to assay variability, which is poorly understood to date. Detailed physicochemical analyses of microplate well surfaces provided new insights into immunoassay variability attributable to microplate surface properties and preparation processes. Our work prompts further investigations to elucidate surface parameters that influence antibody adsorption and biofunctionality, highlighting manufacturing issues that require further review to overcome problems with microplate variability and assay performance.

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Devices for Sample Preparation

2022

Devices and Systems for Laboratory Automation

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A direct heating model to overcome the edge effect in microplates

Cited by 9 publications

References 23 publications

Enzyme activity deviates due to spatial and temporal temperature profiles in commercial microtiter plate readers

Enzyme activity deviates due to spatial and temporal temperature profiles in commercial microtiter plate readers

Variability in Microplate Surface Properties and Its Impact on ELISA

Devices for Sample Preparation

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