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Silver nanoparticle-induced degranulation observed with quantitative phase microscopy

机译：定量相显微镜观察银纳米颗粒诱导的脱粒

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摘要

The use of AgNP is becoming more and more widespread in biomedical field. But compared with the promising bactericidal function, other physiological effects of AgNP on cells are relatively scant. In this research, we propose quantitative phase microscopy (QPM) as a new method to study the degranulation, and AgNP-induced RBL-2H3 cell degranulation is studied as well. Firstly, HeLa cells as the cell control and PBS as the solvent control, we measured the cell volume and cross section profile (x-z plane) with QPM. The results showed that the volume and cross section profile changed only the RBL-2H3 cells exposed to calcium ionophore A23187, which demonstrates the validity of QPM in degranulation research. Secondly, 50μg/mL of AgNP was used instead of A23187, and the measurement of cell volume and cross section profile was carried out again. RBL-2H3 cell volume increased immediately after AgNP was added, and cross section profile showed that the cell surface became granulated, but HeLa cell was lack of that effect. Phase images obviously indicated the RBL-2H3 cell deformation. Thirdly, stained with Fluo-3/AM, intraccllular calcium ([Ca~(2+)]_I ) of single RBL-2H3 cell treated with AgNP was observed with fluorescent microscopy; incubated with AgNP for 20min, thernsupernatant of RBL-2H3 cells was collected and reacted with o-phthalaldehyde (OPA), then the fluorescent intensity ofrnhistamine-OPA complex was assayed with spectrofluorometer. The results of [Ca~(2+)]_I and histaminc increase showed that degranulation of AgNP-induced RBL-2H3 cell occurred. So, the cell volume was used as a parameter of degranulation in our study and AgNP-induced RBL-2H3 cells degranulation was confirmed by the cell volume increment, cross section profile change, and [Ca~(2+)]_I and histamine in supernatant increase.
机译：AgNP的使用在生物医学领域越来越广泛。但是，与有希望的杀菌功能相比，AgNP对细胞的其他生理作用相对较少。在这项研究中，我们提出了定量相显微镜（QPM）作为研究脱颗粒的新方法，并且还研究了AgNP诱导的RBL-2H3细胞脱颗粒。首先，以HeLa细胞为细胞对照，以PBS为溶剂对照，我们用QPM测量细胞体积和横截面轮廓（x-z平面）。结果表明，体积和横截面轮廓仅改变了暴露于钙离子载体A23187的RBL-2H3细胞，这证明了QPM在脱粒研究中的有效性。其次，使用50μg/ mL的AgNP代替A23187，并再次进行细胞体积和横截面轮廓的测量。加入AgNP后，RBL-2H3细胞体积立即增加，截面图显示细胞表面已成粒，但HeLa细胞缺乏这种作用。相图像明显表明RBL-2H3细胞变形。第三，用Fluo-3 / AM染色，用荧光显微镜观察单个AgBL处理的RBL-2H3细胞的细胞内钙（[Ca〜（2 +）] _ I）。用AgNP孵育20min，收集RBL-2H3细胞上清液并与邻苯二甲醛（OPA）反应，然后用荧光分光光度计测定组胺-OPA复合物的荧光强度。 [Ca〜（2 +）] _ I和组蛋白增加的结果表明，AgNP诱导的RBL-2H3细胞发生了脱颗粒。因此，在我们的研究中，将细胞体积用作脱颗粒的参数，并通过细胞体积增加，横截面轮廓变化以及[Ca〜（2 +）] _ I和组胺确定AgNP诱导的RBL-2H3细胞脱颗粒。上清液增加。

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  • 来源
    《Colloidal quantum dots for biomedical applications V》|2010年|P.757515.1-757515.12|共12页
  • 会议地点 San Francisco CA(US)
  • 作者

    Wenzhong Yang; Seungrag Lee; Jiyong Lee; Yoonsung Bae; Dugyoung Kim;

  • 作者单位

    Department of Information and Communications, Gwangju Institute of Science and Technology 1 Oryong-dong, Buk-gu, Gwangju 500-712, Republic of Korea;

    Department of Information and Communications, Gwangju Institute of Science and Technology 1 Oryong-dong, Buk-gu, Gwangju 500-712, Republic of Korea;

    Department of Information and Communications, Gwangju Institute of Science and Technology 1 Oryong-dong, Buk-gu, Gwangju 500-712, Republic of Korea;

    Department of Information and Communications, Gwangju Institute of Science and Technology 1 Oryong-dong, Buk-gu, Gwangju 500-712, Republic of Korea;

    Department of Information and Communications, Gwangju Institute of Science and Technology 1 Oryong-dong, Buk-gu, Gwangju 500-712, Republic of Korea;

  • 会议组织
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 医用物理学;
  • 关键词

    RBL-2H3 cell; degranulation; Silver nanoparticle (AgNP); cell volume; cross section profile; quantitative phase microscopy;

    机译：RBL-2H3细胞；脱粒银纳米粒子（AgNP）；细胞体积横截面轮廓定量相显微镜;

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