Bivariate flow cytometric analysis and sorting of different types of maize starch grains

Zhang, Xudong; Feng, Jiaojiao; Wang, Heng; Zhu, Jiangjiang; Zhong, Yuyue; Liu, Linsan; Xu, Shutu; Zhang, Renhe; Zhang, Xinghua; Xue, Jiquan; Guo, Dongwei

doi:10.1002/cyto.a.23261

Cited by 22 publications

(16 citation statements)

References 26 publications

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“…Currently, flow cytometry is widely considered as an effective method for the classification of granules. So far, this method has only been used in corn starch [12]. Side-scattered light (SSC), forward-scattered light (FSC), and 1-aminopyrene-3,6,8-trisulfonic acid (APTS) represent the integral structure complexity, granule size, and fluorescence intensity (Figure 2), respectively.…”

Section: Resultsmentioning

confidence: 99%

“…The distribution of granule size was determined using Mastersizer 2000 [25]. The complexity and size of starch granule were obtained using flow cytometry (BD FACSAria™ III, Franklin Lakes, NJ, USA), according to the method of Zhang et al [12] with some modifications (the suspension was replaced with double distilled water and the unstained starch granules were used as negative control).…”

Section: Methodsmentioning

confidence: 99%

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Physicochemical Properties of Starches in Proso (Non-Waxy and Waxy) and Foxtail Millets (Non-Waxy and Waxy)

Yang

Zhang

et al. 2019

Molecules

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Proso and foxtail millets are widely cultivated due to their excellent resistance to biotic and abiotic stresses and high nutritional value. Starch is the most important component of millet kernels. Starches with different amylose contents have different physicochemical properties. In this study, starches in proso (non-waxy and waxy) and foxtail millets (non-waxy and waxy) were isolated and investigated. All the starch granules had regular polygonal round shapes and exhibited typical “Maltese crosses”. These four starches all showed bimodal size distribution. The waxy proso and foxtail millets had higher weight-average molar mass and branching degree and lower average chain length of amylopectin. These four starches all presented A-type crystallinity; however, the relative crystallinity of waxy proso and foxtail millets was higher. The two waxy millets had higher onset temperature, peak temperature, conclusion temperature, and gelatinization enthalpy. However, the two non-waxy millets had higher setback viscosity, peak time, and pasting temperature. The significantly different physicochemical properties of waxy and non-waxy millet starches resulted in their different functional properties.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Physicochemical Properties of Starches in Proso (Non-Waxy and Waxy) and Foxtail Millets (Non-Waxy and Waxy)

Yang

Zhang

et al. 2019

Molecules

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show abstract

“…Therefore, the electrical sensing zone method is unaffected by the particle color, shape, composition, or refractive index. Also flow cytometric analyses of starch granules have been reported [91][92][93].…”

Section: Analysis Of Entire Starch Particles and Glycogen Moleculesmentioning

confidence: 99%

Starch and Glycogen Analyses: Methods and Techniques

2020

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For complex carbohydrates, such as glycogen and starch, various analytical methods and techniques exist allowing the detailed characterization of these storage carbohydrates. In this article, we give a brief overview of the most frequently used methods, techniques, and results. Furthermore, we give insights in the isolation, purification, and fragmentation of both starch and glycogen. An overview of the different structural levels of the glucans is given and the corresponding analytical techniques are discussed. Moreover, future perspectives of the analytical needs and the challenges of the currently developing scientific questions are included.

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“…This method functions not only as a technology for isolating target cells but can also provide abundant information on targeted cells, which can help to guide subsequent experiments (2)(3)(4). Cell sorting is indispensable because heterogeneous cell suspensions can be puri ed into fractions containing single cell types based upon virtually unlimited combinations of user-de ned parameters (5). Therefore, the cell sorter is the foundation for numerous downstream studies.…”

Section: Introductionmentioning

confidence: 99%

Improved Strategy for Jet-in-air Cell Sorting With High Purity, Yield, Viability and Genome Stability

Song

Wang

et al. 2020

Preprint

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Background: The enrichment of viable cells is an essential step in single-cell sequencing for clinical and biotechnology research applications. A high-end jet-in-air sorter is a fluorescent-activated cell sorting sorter that ideally processes cells with high purity, yield and viability. We want to improve the strategy for jet-in-air cell sorting to get more effective cells, and ensure the downstream experiments. Method: In this study, to achieve high-purity, high-yield, high-viability cell sorting, we explored three aspects. First, we manually adjusted the drop delay through the beads experiment to make it more accurate and to ensure the highest yield in theory, thereby avoiding the influence of the instrument and operator. Second, the effects on cell apoptosis caused by adding 2% fetal bovine serum to the loading buffer were examined. Third, the effects on cell count, apoptosis, proliferation and gene expression caused by employing FBS and bovine serum albumin in the collecting buffer were analyzed. Statistical analysis was performed with GraphPad Prism 6 software. Significance was set at p < 0.05.Result: Our data showed that an additional manual correction of drop delay could lead to an improved cell yield of 98.28%±3.79%. Adding FBS (2%) to the loading buffer had no significant effect on the fate of sorted cells in 4 hours. However, employing a suitable concentration of FBS/BSA in the collecting buffer mediated a notable increase in cell count from 50% to ~100%, a significant decrease in cell apoptosis from ~40% to less than 10% for cell lines and ~70% to less than 40% for CD45+ primary cells from mouse spleen. And a clear increase in cell proliferation at 48 h after sorting for jurkat cells. Moreover, the level of gene expression remained steady in the 5% FBS collecting buffer. Conclusions: Our manuscript demonstrates techniques that can be easily followed to refine sort yields of healthy cells. Fluorescent-activated cell sorting users could easily obtain enough target cells with great viability and need not to worry about any problems related to collecting or loading buffer. Furthermore, adding extra expensive FBS in collecting or loading buffer to boost yield is no longer necessary. In all, this will make the downstream experiments smoother.

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Bivariate flow cytometric analysis and sorting of different types of maize starch grains

Cited by 22 publications

References 26 publications

Physicochemical Properties of Starches in Proso (Non-Waxy and Waxy) and Foxtail Millets (Non-Waxy and Waxy)

Physicochemical Properties of Starches in Proso (Non-Waxy and Waxy) and Foxtail Millets (Non-Waxy and Waxy)

Starch and Glycogen Analyses: Methods and Techniques

Improved Strategy for Jet-in-air Cell Sorting With High Purity, Yield, Viability and Genome Stability

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