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Distinct deposition of amyloid-bspecies in brains with Alzheimer’s disease pathology visualized with MALDI imaging mass spectrometry
EP29713
Poster Title: Distinct deposition of amyloid-bspecies in brains with Alzheimer’s disease pathology visualized with MALDI imaging mass spectrometry
Submitted on 13 Feb 2019
Author(s): Nobuto Kakuda1, Tomohiro Miyasaka1, Takashi Nirasawa2, Shigeo Murayama3, Yasuo Ihara1 and Masaya Ikegawa1
Affiliations: 1.Doshisha University, Faculty of Life and Medical Systems, Kyoto, Japan 2.Bruker Japan, K.K., Yokohama, Japan 3.The Brain Bank for Aging Research, Tokyo Metropolitan Geriatric Hospital, Itabashi, Japan
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Abstract: Subjects: Human cortical specimens for IMS and immunohistochemistry were obtained from those brains that were removed processed and placed in -80˚C within 8h postmortem at the Brain bank at Tokyo Metropolitan Institute of Gerontology. For all brains registered at the brain bank we obtained written informed consents for their use for medical research from patients or patient’s family. Each brain specimen was taken from occipital cortex of 5 AD patients and 5 controls.

MALDI Imaging: Frozen tissue sections were cut on a cryostat (CM1950, Leica Microsystems, Wetzlar, Germany) at a 10mm thickness onto indium-tin-oxide–coated glass slides (Bruker Daltonics, Bremen, Germany). Before matrix coating, treated with a formic acid. For mass spectrometric measurements, tissue areas were defined using the FlexControl 3.8 and Flex Imaging 5.0 software packages (both Bruker Daltonics). Spectra were acquired using the rapifleX MALDI Tissuetyper (Bruker Daltonics) in positive linear mode, whereas ions were detected in a mass range of m/z 2,000 to 20,000 with spatial resolution of 20, 50 and 100mm. A ready-made protein standard was used for spectra calibration (Bruker Daltonics). Visualization and statistical analysis were used Flex Imaging and SciLS Lab 2016a (SCiLS, Bremen, Germany).

Immunohistochemistry: Fresh frozen sections were post- fixed by 4% paraformaldehyde (PFA) in phosphate buffered saline (PBS). After rinses with PBS-T, bound antibodies were visualized with secondary antibodies conjugated with Alexa dyes (life technology). The specimens were analyzed by confocal-laser-scanning-microscope (LSM700; Carl Zeiss Inc.)
Summary: Here we demonstrate a detailed analysis of the different variants of Ab peptides in the brains from patients with and without AD pathology including a variety of cerebral amyloid angiopathy (CAA).References: Kakuda N and MiyasakaT et al (2017) Distinct deposition of amyloid-β species in brains with Alzheimer’s disease pathology visualized with MALDI imaging mass spectrometry. ActaNeuropathologicaComm. 5: 73. Oct 16.Report abuse »
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