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Proceedings Paper

Silver nanoparticle-induced degranulation observed with quantitative phase microscopy
Author(s): Wenzhong Yang; Seungrag Lee; Jiyong Lee; Yoonsung Bae; Dugyoung Kim
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Paper Abstract

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, intracellular calcium Ca2+]i of single RBL-2H3 cell treated with AgNP was observed with fluorescent microscopy; incubated with AgNP for 20min, the supernatant of RBL-2H3 cells was collected and reacted with o-phthalaldehyde (OPA), then the fluorescent intensity of histamine-OPA complex was assayed with spectrofluorometer. The results of Ca2+]i and histamine 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 [Ca2+]i and histamine in supernatant increase.

Paper Details

Date Published: 18 February 2010
PDF: 12 pages
Proc. SPIE 7575, Colloidal Quantum Dots for Biomedical Applications V, 757515 (18 February 2010); doi: 10.1117/12.842925
Show Author Affiliations
Wenzhong Yang, Gwangju Institute of Science and Technology (Korea, Republic of)
Seungrag Lee, Gwangju Institute of Science and Technology (Korea, Republic of)
Jiyong Lee, Gwangju Institute of Science and Technology (Korea, Republic of)
Yoonsung Bae, Gwangju Institute of Science and Technology (Korea, Republic of)
Dugyoung Kim, Gwangju Institute of Science and Technology (Korea, Republic of)


Published in SPIE Proceedings Vol. 7575:
Colloidal Quantum Dots for Biomedical Applications V
Marek Osinski; Wolfgang J. Parak; Thomas M. Jovin; Kenji Yamamoto, Editor(s)

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