PMID- 30390005
OWN - NLM
STAT- MEDLINE
DCOM- 20191119
LR  - 20191119
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 8
IP  - 1
DP  - 2018 Nov 2
TI  - Flow cytometric analysis of Xenopus laevis and X. tropicalis blood cells using 
      acridine orange.
PG  - 16245
LID - 10.1038/s41598-018-34631-0 [doi]
LID - 16245
AB  - 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.
FAU - Sato, Kei
AU  - Sato K
AUID- ORCID: 0000-0001-6923-5594
AD  - Faculty of Education and Integrated Arts and Sciences, Waseda University, 2-2 
      Wakamatsu, Shinjuku, Tokyo, 162-8480, Japan.
AD  - Major in Integrative Bioscience and Biomedical Engineering, Graduate School of 
      Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu, Shinjuku, 
      Tokyo, 162-8480, Japan.
FAU - Uehara, Azusa
AU  - Uehara A
AD  - Major in Integrative Bioscience and Biomedical Engineering, Graduate School of 
      Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu, Shinjuku, 
      Tokyo, 162-8480, Japan.
FAU - Kinoshita, Sayaka
AU  - Kinoshita S
AD  - Major in Integrative Bioscience and Biomedical Engineering, Graduate School of 
      Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu, Shinjuku, 
      Tokyo, 162-8480, Japan.
FAU - Nomura, Ikki
AU  - Nomura I
AD  - Major in Integrative Bioscience and Biomedical Engineering, Graduate School of 
      Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu, Shinjuku, 
      Tokyo, 162-8480, Japan.
FAU - Yagi, Minami
AU  - Yagi M
AD  - Major in Integrative Bioscience and Biomedical Engineering, Graduate School of 
      Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu, Shinjuku, 
      Tokyo, 162-8480, Japan.
FAU - Tanizaki, Yuta
AU  - Tanizaki Y
AD  - Major in Integrative Bioscience and Biomedical Engineering, Graduate School of 
      Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu, Shinjuku, 
      Tokyo, 162-8480, Japan.
FAU - Matsuda-Shoji, Yu
AU  - Matsuda-Shoji Y
AD  - Major in Integrative Bioscience and Biomedical Engineering, Graduate School of 
      Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu, Shinjuku, 
      Tokyo, 162-8480, Japan.
FAU - Matsubayashi, Atsushi
AU  - Matsubayashi A
AD  - Major in Integrative Bioscience and Biomedical Engineering, Graduate School of 
      Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu, Shinjuku, 
      Tokyo, 162-8480, Japan.
FAU - Endo, Nobuyasu
AU  - Endo N
AD  - Faculty of Education and Integrated Arts and Sciences, Waseda University, 2-2 
      Wakamatsu, Shinjuku, Tokyo, 162-8480, Japan.
FAU - Nagai, Yutaka
AU  - Nagai Y
AD  - Technology Center, Nihon Kohden Corporation, 1-31-4, Nishiochiai, Shinjuku, 
      Tokyo, Japan.
FAU - Kato, Takashi
AU  - Kato T
AUID- ORCID: 0000-0003-0384-7971
AD  - Faculty of Education and Integrated Arts and Sciences, Waseda University, 2-2 
      Wakamatsu, Shinjuku, Tokyo, 162-8480, Japan. tkato@waseda.jp.
AD  - Major in Integrative Bioscience and Biomedical Engineering, Graduate School of 
      Advanced Science and Engineering, Waseda University, 2-2 Wakamatsu, Shinjuku, 
      Tokyo, 162-8480, Japan. tkato@waseda.jp.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20181102
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Fluorescent Dyes)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/chemistry
MH  - Animals
MH  - Animals, Inbred Strains/blood
MH  - Animals, Wild/blood
MH  - Blood Cell Count/methods
MH  - *Blood Cells
MH  - Cell Separation/*methods
MH  - Flow Cytometry/*methods
MH  - Fluorescent Dyes/chemistry
MH  - Models, Animal
MH  - Species Specificity
MH  - Staining and Labeling/*methods
MH  - Xenopus laevis/*blood
PMC - PMC6214894
COIS- Y. N. is an employee of NIHON KOHDEN CORPORATION.
EDAT- 2018/11/06 06:00
MHDA- 2019/11/20 06:00
PMCR- 2018/11/02
CRDT- 2018/11/04 06:00
PHST- 2018/05/08 00:00 [received]
PHST- 2018/10/01 00:00 [accepted]
PHST- 2018/11/04 06:00 [entrez]
PHST- 2018/11/06 06:00 [pubmed]
PHST- 2019/11/20 06:00 [medline]
PHST- 2018/11/02 00:00 [pmc-release]
AID - 10.1038/s41598-018-34631-0 [pii]
AID - 34631 [pii]
AID - 10.1038/s41598-018-34631-0 [doi]
PST - epublish
SO  - Sci Rep. 2018 Nov 2;8(1):16245. doi: 10.1038/s41598-018-34631-0.