Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function

Gulnaz Stybayeva, Onur Mudanyali, Sungkyu Seo, Jaime Silangcruz, Monica MacAl, Erlan Ramanculov, Satya Dandekar, Anthony Erlinger, Aydogan Ozcan, Alexander Revzin

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Characterization of leukocytes is an integral part of blood analysis and blood-based diagnostics. In the present paper, we combine lensless holographic imaging with antibody microarrays for rapid and multiparametric analysis of leukocytes from human blood. Monoclonal antibodies (Abs) specific for leukocyte surface antigens (CD4 and CD8) and cytokines (TNF-α, IFN-γ, IL-2) were printed in an array so as to juxtapose cell capture and cytokine detection antibody (Ab) spots. Integration of Ab microarrays into a microfluidic flow chamber (4 μL volume) followed by incubation with human blood resulted in capture of CD4 and CD8 T-cells on specific Ab spots. On-chip mitogenic activation of these cells induced release of cytokine molecules that were subsequently captured on neighboring anticytokine Ab spots. The binding of IL-2, TNF-α, and IFN-γ molecules on their respective Ab spots was detected using horseradish peroxidase (HRP)-labeled anticytokine Abs and a visible color reagent. Lensfree holographic imaging was then used to rapidly (∼4 s) enumerate CD4 and CD8 T-lymphocytes captured on Ab spots and to quantify the cytokine signal emanating from IL-2, TNF-α, and IFN-γ spots on the same chip. To demonstrate the utility of our approach for infectious disease monitoring, blood samples of healthy volunteers and human immunodeficiency virus (HIV)-infected patients were analyzed to determine the CD4/CD8 ratio, an important HIV/AIDS diagnostic marker. The ratio obtained by lensfree on-chip imaging of CD4 and CD8 T-cells captured on Ab spots was in close agreement with conventional microscopy-based cell counting. The present paper, describing tandem use of Ab microarrays and lensfree holographic imaging, paves the way for future development of miniature cytometry devices for multiparametric blood analysis at the point of care or in a resource-limited setting.

Original languageEnglish
Pages (from-to)3736-3744
Number of pages9
JournalAnalytical Chemistry
Volume82
Issue number9
DOIs
Publication statusPublished - May 1 2010
Externally publishedYes

Fingerprint

Microarrays
Throughput
Imaging techniques
Antibodies
Blood
T-cells
Cytokines
Interleukin-2
Viruses
Molecules
Horseradish Peroxidase
Surface Antigens
Microfluidics
Microscopic examination
Chemical activation
Monoclonal Antibodies
Color
Monitoring

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function. / Stybayeva, Gulnaz; Mudanyali, Onur; Seo, Sungkyu; Silangcruz, Jaime; MacAl, Monica; Ramanculov, Erlan; Dandekar, Satya; Erlinger, Anthony; Ozcan, Aydogan; Revzin, Alexander.

In: Analytical Chemistry, Vol. 82, No. 9, 01.05.2010, p. 3736-3744.

Research output: Contribution to journalArticle

Stybayeva, G, Mudanyali, O, Seo, S, Silangcruz, J, MacAl, M, Ramanculov, E, Dandekar, S, Erlinger, A, Ozcan, A & Revzin, A 2010, 'Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function', Analytical Chemistry, vol. 82, no. 9, pp. 3736-3744. https://doi.org/10.1021/ac100142a
Stybayeva, Gulnaz ; Mudanyali, Onur ; Seo, Sungkyu ; Silangcruz, Jaime ; MacAl, Monica ; Ramanculov, Erlan ; Dandekar, Satya ; Erlinger, Anthony ; Ozcan, Aydogan ; Revzin, Alexander. / Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function. In: Analytical Chemistry. 2010 ; Vol. 82, No. 9. pp. 3736-3744.
@article{25949987fe7c456ca251779fb53e86ea,
title = "Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function",
abstract = "Characterization of leukocytes is an integral part of blood analysis and blood-based diagnostics. In the present paper, we combine lensless holographic imaging with antibody microarrays for rapid and multiparametric analysis of leukocytes from human blood. Monoclonal antibodies (Abs) specific for leukocyte surface antigens (CD4 and CD8) and cytokines (TNF-α, IFN-γ, IL-2) were printed in an array so as to juxtapose cell capture and cytokine detection antibody (Ab) spots. Integration of Ab microarrays into a microfluidic flow chamber (4 μL volume) followed by incubation with human blood resulted in capture of CD4 and CD8 T-cells on specific Ab spots. On-chip mitogenic activation of these cells induced release of cytokine molecules that were subsequently captured on neighboring anticytokine Ab spots. The binding of IL-2, TNF-α, and IFN-γ molecules on their respective Ab spots was detected using horseradish peroxidase (HRP)-labeled anticytokine Abs and a visible color reagent. Lensfree holographic imaging was then used to rapidly (∼4 s) enumerate CD4 and CD8 T-lymphocytes captured on Ab spots and to quantify the cytokine signal emanating from IL-2, TNF-α, and IFN-γ spots on the same chip. To demonstrate the utility of our approach for infectious disease monitoring, blood samples of healthy volunteers and human immunodeficiency virus (HIV)-infected patients were analyzed to determine the CD4/CD8 ratio, an important HIV/AIDS diagnostic marker. The ratio obtained by lensfree on-chip imaging of CD4 and CD8 T-cells captured on Ab spots was in close agreement with conventional microscopy-based cell counting. The present paper, describing tandem use of Ab microarrays and lensfree holographic imaging, paves the way for future development of miniature cytometry devices for multiparametric blood analysis at the point of care or in a resource-limited setting.",
author = "Gulnaz Stybayeva and Onur Mudanyali and Sungkyu Seo and Jaime Silangcruz and Monica MacAl and Erlan Ramanculov and Satya Dandekar and Anthony Erlinger and Aydogan Ozcan and Alexander Revzin",
year = "2010",
month = "5",
day = "1",
doi = "10.1021/ac100142a",
language = "English",
volume = "82",
pages = "3736--3744",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "9",

}

TY - JOUR

T1 - Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function

AU - Stybayeva, Gulnaz

AU - Mudanyali, Onur

AU - Seo, Sungkyu

AU - Silangcruz, Jaime

AU - MacAl, Monica

AU - Ramanculov, Erlan

AU - Dandekar, Satya

AU - Erlinger, Anthony

AU - Ozcan, Aydogan

AU - Revzin, Alexander

PY - 2010/5/1

Y1 - 2010/5/1

N2 - Characterization of leukocytes is an integral part of blood analysis and blood-based diagnostics. In the present paper, we combine lensless holographic imaging with antibody microarrays for rapid and multiparametric analysis of leukocytes from human blood. Monoclonal antibodies (Abs) specific for leukocyte surface antigens (CD4 and CD8) and cytokines (TNF-α, IFN-γ, IL-2) were printed in an array so as to juxtapose cell capture and cytokine detection antibody (Ab) spots. Integration of Ab microarrays into a microfluidic flow chamber (4 μL volume) followed by incubation with human blood resulted in capture of CD4 and CD8 T-cells on specific Ab spots. On-chip mitogenic activation of these cells induced release of cytokine molecules that were subsequently captured on neighboring anticytokine Ab spots. The binding of IL-2, TNF-α, and IFN-γ molecules on their respective Ab spots was detected using horseradish peroxidase (HRP)-labeled anticytokine Abs and a visible color reagent. Lensfree holographic imaging was then used to rapidly (∼4 s) enumerate CD4 and CD8 T-lymphocytes captured on Ab spots and to quantify the cytokine signal emanating from IL-2, TNF-α, and IFN-γ spots on the same chip. To demonstrate the utility of our approach for infectious disease monitoring, blood samples of healthy volunteers and human immunodeficiency virus (HIV)-infected patients were analyzed to determine the CD4/CD8 ratio, an important HIV/AIDS diagnostic marker. The ratio obtained by lensfree on-chip imaging of CD4 and CD8 T-cells captured on Ab spots was in close agreement with conventional microscopy-based cell counting. The present paper, describing tandem use of Ab microarrays and lensfree holographic imaging, paves the way for future development of miniature cytometry devices for multiparametric blood analysis at the point of care or in a resource-limited setting.

AB - Characterization of leukocytes is an integral part of blood analysis and blood-based diagnostics. In the present paper, we combine lensless holographic imaging with antibody microarrays for rapid and multiparametric analysis of leukocytes from human blood. Monoclonal antibodies (Abs) specific for leukocyte surface antigens (CD4 and CD8) and cytokines (TNF-α, IFN-γ, IL-2) were printed in an array so as to juxtapose cell capture and cytokine detection antibody (Ab) spots. Integration of Ab microarrays into a microfluidic flow chamber (4 μL volume) followed by incubation with human blood resulted in capture of CD4 and CD8 T-cells on specific Ab spots. On-chip mitogenic activation of these cells induced release of cytokine molecules that were subsequently captured on neighboring anticytokine Ab spots. The binding of IL-2, TNF-α, and IFN-γ molecules on their respective Ab spots was detected using horseradish peroxidase (HRP)-labeled anticytokine Abs and a visible color reagent. Lensfree holographic imaging was then used to rapidly (∼4 s) enumerate CD4 and CD8 T-lymphocytes captured on Ab spots and to quantify the cytokine signal emanating from IL-2, TNF-α, and IFN-γ spots on the same chip. To demonstrate the utility of our approach for infectious disease monitoring, blood samples of healthy volunteers and human immunodeficiency virus (HIV)-infected patients were analyzed to determine the CD4/CD8 ratio, an important HIV/AIDS diagnostic marker. The ratio obtained by lensfree on-chip imaging of CD4 and CD8 T-cells captured on Ab spots was in close agreement with conventional microscopy-based cell counting. The present paper, describing tandem use of Ab microarrays and lensfree holographic imaging, paves the way for future development of miniature cytometry devices for multiparametric blood analysis at the point of care or in a resource-limited setting.

UR - http://www.scopus.com/inward/record.url?scp=77951861838&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77951861838&partnerID=8YFLogxK

U2 - 10.1021/ac100142a

DO - 10.1021/ac100142a

M3 - Article

VL - 82

SP - 3736

EP - 3744

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 9

ER -