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Biomed Opt Express. 2015 Aug 7;6(9):3303-12. doi: 10.1364/BOE.6.003303. eCollection 2015 Sep 1.

In vivo imaging of activated microglia in a mouse model of focal cerebral ischemia by two-photon microscopy.

Author information

1
Division of Integrative Biosciences and Biotechnology, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-Gu, Pohang, Gyeongbuk 790-784, South Korea.
2
Department of Chemistry, Pohang University of Science and Technology, 77 Cheongam-Ro, Nam-Gu, Pohang, Gyeongbuk 790-784, South Korea.
3
Department of Neurology, Seoul National University College of Medicine, 103 Daehak-Ro, Jongno-Gu, Seoul 110-799, South Korea.
4
Department of Radiation Oncology, Seoul National University College of Medicine, 103 Daehak-Ro, Jongno-Gu, Seoul 110-799, South Korea.

Abstract

Microglia are brain resident macrophages rapidly responding to various stimuli to exert appropriate inflammatory responses. Although they have recently been exploited as an attractive candidate for imaging neuroinflammation, it is still difficult to visualize them at the cellular and molecular levels. Here we imaged activated microglia by establishing intracranial window chamber (ICW) in a mouse model of focal cerebral ischemia by using two-photon microscopy (TPM), in vivo. Intravenous injection of fluorescent antibodies allowed us to detect significantly elevated levels of Iba-1 and CD68 positive activated microglia in the ipsilateral compared to the contralateral side of the infarct. We further observed that indomethacin, a non-steroidal anti-inflammatory drug significantly attenuated CD68-positive microglial activation in ICW, which was further confirmed by qRT-PCR biochemical analyses. In conclusion, we believe that in vivo TPM imaging of ICW would be a useful tool to screen for therapeutic interventions lowering microglial activation hence neuroinflammation.

KEYWORDS:

(000.1430) Biology and medicine; (180.2520) Fluorescence microscopy; (180.4315) Nonlinear microscopy

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