Flow cytometric analysis of Xenopus laevis and X. tropicalis blood cells using acridine orange

Kei Sato, Azusa Uehara, Sayaka Kinoshita, Ikki Nomura, Minami Yagi, Yuta Tanizaki, Yu Matsuda-shoji, Atsushi Matsubayashi, Nobuyasu Endo, Yutaka Nagai, Takashi Kato

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2 Citations (Scopus)

Abstract

Automated blood cell counters can distinguish cells based on their size and the presence or absence of a nucleus. However, most vertebrates have nucleated blood cells that cannot be counted automatically. We established an alternative automatic method for counting peripheral blood cells by staining cells with the fluorescent dye acridine orange (AO) and analysing cell populations using flow cytometry (FCM). As promising new animal models, we chose Xenopus laevis and three inbred strains of X. tropicalis. We compared the haematological phenotypes, including blood cell types, cell sizes, cellular structure, and erythrocyte lifespans/turnover rate among X. laevis and the three inbred strains of X. tropicalis. Each cell type from X. laevis was sorted according to six parameters: forward- and side-scattered light emission, AO red and green fluorescence intensity, and cellular red and green fluorescence. Remarkably, the erythrocyte count was the highest in the Golden line, suggesting that genetic factors were associated with the blood cells. Furthermore, immature erythrocytes in anaemic X. laevis could be separated from normal blood cells based on red fluorescence intensity. These results show that FCM with AO staining allows for an accurate analysis of peripheral blood cells from various species.

Original languageEnglish
Article number16245
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - 2018 Dec 1

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Acridine Orange
Xenopus laevis
Blood Cells
Fluorescence
Flow Cytometry
Erythrocytes
Staining and Labeling
Erythrocyte Count
Cellular Structures
Fluorescent Dyes
Cell Size
Vertebrates
Animal Models
Phenotype
Light
Population

ASJC Scopus subject areas

  • General

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Flow cytometric analysis of Xenopus laevis and X. tropicalis blood cells using acridine orange. / Sato, Kei; Uehara, Azusa; Kinoshita, Sayaka; Nomura, Ikki; Yagi, Minami; Tanizaki, Yuta; Matsuda-shoji, Yu; Matsubayashi, Atsushi; Endo, Nobuyasu; Nagai, Yutaka; Kato, Takashi.

In: Scientific Reports, Vol. 8, No. 1, 16245, 01.12.2018.

Research output: Contribution to journalArticle

Sato, K, Uehara, A, Kinoshita, S, Nomura, I, Yagi, M, Tanizaki, Y, Matsuda-shoji, Y, Matsubayashi, A, Endo, N, Nagai, Y & Kato, T 2018, 'Flow cytometric analysis of Xenopus laevis and X. tropicalis blood cells using acridine orange', Scientific Reports, vol. 8, no. 1, 16245. https://doi.org/10.1038/s41598-018-34631-0
Sato, Kei ; Uehara, Azusa ; Kinoshita, Sayaka ; Nomura, Ikki ; Yagi, Minami ; Tanizaki, Yuta ; Matsuda-shoji, Yu ; Matsubayashi, Atsushi ; Endo, Nobuyasu ; Nagai, Yutaka ; Kato, Takashi. / Flow cytometric analysis of Xenopus laevis and X. tropicalis blood cells using acridine orange. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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