Abstract
Absolute counting of total leukocytes or specific subsets within small amounts of whole blood is difficult due to a lack of techniques that enable separation of all leukocytes from limited amounts of whole blood. In this study, a microfluidic device equipped with a size-controlled microcavity array for highly efficient separation of leukocytes from submicroliters of whole blood was developed. The microcavity array can separate leukocytes from whole blood based on differences in the size and deformability between leukocytes and other blood cells. Leukocytes recovered on aligned microcavities were continuously processed for image-based immunophenotypic analysis. Our device successfully recovered over 90% of leukocytes in 1μL of whole blood without pretreatment such as density gradient centrifugation or erythrocyte lysis. In addition, the proposed system successfully performed absolute enumeration of human CD4 + and CD8 + leukocytes from 1μL of whole blood, and the obtained data showed good correlation with conventional flow cytometric analysis. Our microfluidic device has great potential as a tool for a point-of-care leukocyte analysis system.
| Original language | English |
|---|---|
| Pages (from-to) | 2017-2024 |
| Number of pages | 8 |
| Journal | Biotechnology and Bioengineering |
| Volume | 109 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 2012 Aug |
| Externally published | Yes |
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Keywords
- CD4/CD8
- Cell separation
- Electroforming
- Leukocyte
- Microfluidic device
- Whole blood
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology
Cite this
Leukocyte counting from a small amount of whole blood using a size-controlled microcavity array. / Hosokawa, Masahito; Asami, Marie; Nakamura, Seita; Yoshino, Tomoko; Tsujimura, Noriyuki; Takahashi, Masayuki; Nakasono, Satoshi; Tanaka, Tsuyoshi; Matsunaga, Tadashi.
In: Biotechnology and Bioengineering, Vol. 109, No. 8, 08.2012, p. 2017-2024.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Leukocyte counting from a small amount of whole blood using a size-controlled microcavity array
AU - Hosokawa, Masahito
AU - Asami, Marie
AU - Nakamura, Seita
AU - Yoshino, Tomoko
AU - Tsujimura, Noriyuki
AU - Takahashi, Masayuki
AU - Nakasono, Satoshi
AU - Tanaka, Tsuyoshi
AU - Matsunaga, Tadashi
PY - 2012/8
Y1 - 2012/8
N2 - Absolute counting of total leukocytes or specific subsets within small amounts of whole blood is difficult due to a lack of techniques that enable separation of all leukocytes from limited amounts of whole blood. In this study, a microfluidic device equipped with a size-controlled microcavity array for highly efficient separation of leukocytes from submicroliters of whole blood was developed. The microcavity array can separate leukocytes from whole blood based on differences in the size and deformability between leukocytes and other blood cells. Leukocytes recovered on aligned microcavities were continuously processed for image-based immunophenotypic analysis. Our device successfully recovered over 90% of leukocytes in 1μL of whole blood without pretreatment such as density gradient centrifugation or erythrocyte lysis. In addition, the proposed system successfully performed absolute enumeration of human CD4 + and CD8 + leukocytes from 1μL of whole blood, and the obtained data showed good correlation with conventional flow cytometric analysis. Our microfluidic device has great potential as a tool for a point-of-care leukocyte analysis system.
AB - Absolute counting of total leukocytes or specific subsets within small amounts of whole blood is difficult due to a lack of techniques that enable separation of all leukocytes from limited amounts of whole blood. In this study, a microfluidic device equipped with a size-controlled microcavity array for highly efficient separation of leukocytes from submicroliters of whole blood was developed. The microcavity array can separate leukocytes from whole blood based on differences in the size and deformability between leukocytes and other blood cells. Leukocytes recovered on aligned microcavities were continuously processed for image-based immunophenotypic analysis. Our device successfully recovered over 90% of leukocytes in 1μL of whole blood without pretreatment such as density gradient centrifugation or erythrocyte lysis. In addition, the proposed system successfully performed absolute enumeration of human CD4 + and CD8 + leukocytes from 1μL of whole blood, and the obtained data showed good correlation with conventional flow cytometric analysis. Our microfluidic device has great potential as a tool for a point-of-care leukocyte analysis system.
KW - CD4/CD8
KW - Cell separation
KW - Electroforming
KW - Leukocyte
KW - Microfluidic device
KW - Whole blood
UR - http://www.scopus.com/inward/record.url?scp=84862824414&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84862824414&partnerID=8YFLogxK
U2 - 10.1002/bit.24471
DO - 10.1002/bit.24471
M3 - Article
C2 - 22367741
AN - SCOPUS:84862824414
VL - 109
SP - 2017
EP - 2024
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
SN - 0006-3592
IS - 8
ER -