Skip to main content
SpringerLink
Log in

A novel integrated microfluidic platform to perform fluorescence in situ hybridization for chromosomal analysis

  • Research Paper
  • Published:
Microfluidics and Nanofluidics Aims and scope Submit manuscript

Abstract

The fluorescence in situ hybridization (FISH) technique has been commonly employed to detect the chromosomal abnormalities. However, applications of this technique are limited due to its lengthy process and labor-intensive sample preparation. In this study, a novel integrated microfluidic chip capable of performing the entire FISH protocol automatically was reported. This novel technique can achieve several advantages, including reduce the consumption of bio-samples and reagents, automation and rapid analysis compared to the conventional method. In this study, several functional microfluidic devices were integrated on a single chip to perform automatic FISH on the microfluidic platform. Experimental data demonstrated that the developed microfluidic system successfully provided superior performance for probing the chromosomal abnormality of cells. Furthermore, the novel microfluidic system performed the entire process automatically within 3 h, where the conventional method required 10 h to perform the entire protocol manually. This data indicated superior performance of the novel method. Our findings conclude that the novel integrated FISH protocol is more convenient to perform large quantities of samples, which can be used in clinical trials.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

Abbreviations

ATCC:

American Type Culture Collection

Bio-MEMS:

Bio-microelectromechanical systems

CNC:

Computer numerical control

DAPI:

4′-Diamidino-2-phenylindole

DC:

Direct current

ddH2O:

Double-distilled water

EDTA-K3:

Ethylenediaminetetraacetic acid tripotassium dihydrate

EMV:

Electromagnetic valve

EtOH:

Ethyl alcohol

FISH:

Fluorescence in situ hybridization

PBMCs:

Peripheral blood mononuclear cells

PBS:

Phosphate-buffered saline

PDMS:

Polydimethylsiloxane

PMMA:

Polymethymethacrylate

SSC:

Sodium chloride–sodium citrate buffer

TE:

Thermoelectric

References

  • Andreeff M, Pinkel D (1999) Introduction to Fluorescence in Situ Hybridization: principles and clinical applications. p 455

  • Chee M, Yang R, Hubbell E, Berno A, Huang XC, Stern D, Winkler J, Lockhart DJ, Morris MS, Fodor SPA (1996) Accessing genetic information with high–density DNA arrays. Science 274:610–614

    Article  Google Scholar 

  • Cho YK, Lee JG, Park JM, Lee BS, Lee Y, Ko C (2007) One-step pathogen specific DNA extraction from whole blood on a centrifugal microfluidic device. Lab Chip 7:565–573

    Article  Google Scholar 

  • Edman CF, Raymond DE, Wu DJ, Tu E, Sosnowski RG, Butler WF, Nerenberg M, Heller MJ (1997) Electric field directed nucleic acid hybridization on microchips. Nucleic Acids Res 25:4907–4914

    Article  Google Scholar 

  • Erickson D, Liu X, Krull U, Li D (2004) Electrokinetically controlled DNA hybridization microfluidic chip enabling rapid target analysis. Anal Chem 76:7269–7277

    Article  Google Scholar 

  • Fox JL, Hsu PH, Legator MS, Morrison LE, Seeling SA (1995) Fluorescence in situ hybridization: powerful molecular tool for cancer prognosis. Clin Chem 41:1554–1559

    Google Scholar 

  • Gu Y, Cimino G, Alder H, Nakamura T, Prasad R, Canaani O, Moirt DT, Jones C, Nowell PC, Croce CM, Canaani E (1992) The (4;11)(q21;q23) chromosome translocations in acute leukemias involve the VDJ recombinase. Genetics 89:10464–10468

    Google Scholar 

  • Huang SF, Xiao S, Renshaw AA, Loughlin KR, Hudson TJ, Fletcher JA (1996) Fluorescence in Situ Hybridization Evaluation of Chromosome Deletion Patterns in Prostate Cancer. American Journal of Patbology 149:1565–1573

    Google Scholar 

  • Kitayama Y, Igarashi H, Kozu T, Nagura K, Ohashi Y, Sugimura H (2006) Repeated fluorescence in situ hybridization by a microwave-enhanced protocol. Pathol Int 56:490–493

    Article  Google Scholar 

  • Langer-Safer PR, Levine M, Ward DC (1982) Immunological method for mapping genes on Drosophila polytene chromosomes. Proc Natl Acad Sci USA 79:4381–4385

    Article  Google Scholar 

  • Leversha MA, Han J, Asgari Z, Danila DC, Lin O, Espinoza RG, Anand A, Lilja H, Heller G, Fleisher M, Scher HL (2009) Fluorescence In situ Hybridization Analysis of Circulating Tumor Cells in Metastatic Prostate Cancer. Clin Cancer Res 15:2091–2097

    Article  Google Scholar 

  • Lichter P, Ledbetter SA, Ledbetter DH, Ward DC (1990) Fluorescence in situ hybridization with Alu and L1 polymerase chain reaction probes for rapid characterization of human chromosomes in hybrid cell lines. Proc Natl Acad Sci USA 87:6634–6638

    Article  Google Scholar 

  • Nath J, Johnson KL (2000) A review of fluorescence in situ hybridization (FISH): current status and future prospects. Biotech Histochem 75:54–78

    Article  Google Scholar 

  • Nusse R, Fuerer C, Ching W, Harnish K, Logan C, Zeng A, Ten Berge D, Kalani Y (2008) Wnt signaling and stem cell control. In: Stillman B, Stewart S, Grodzicker T (eds) Control and regulation of stem cells. Harbor Laboratory Press, Cold Spring, pp 59–66

    Google Scholar 

  • Ochsner M, Textor M, Vogel V, Smith ML (2010) Dimensionality controls cytoskeleton assembly and metabolism of fibroblast cells in response to rigidity and shape. Plos One 5(3)

  • Pinkel D, Landegent J, Collins C, Fuscoe J, Segraves R, Lucas J, Gray J (1988) Fluorescence in situ hybridization with human chromosome-specific libraries: detection of trisomy 21 and translocations of chromosome 4. Proc Natl Acad Sci USA 85:9138–9142

    Article  Google Scholar 

  • Radtkey R, Feng L, Muralhida M, Duhon M, Canter D, DiPierro D, Fallon S, Tu E, McElfresh K, Nerenberg M, Sosnowski R (2000) Rapid, high fidelity analysis of simple sequence repeats on an electronically active DNA microchip. Nucleic Acids Res 28:E17

    Article  Google Scholar 

  • Raj A, Bogaard PVD, Rifkin SA, Oudenaarden AV, Tyagi S (2008) Imaging individual mRNA molecules using multiple singly labeled probes. Nat Methods 5:877–879

    Article  Google Scholar 

  • Sieben VJ, Debes Marun CS, Pilarski PM, Kaigala GV, Pilarski LM, Backhouse CJ (2007) FISH and chips: chromosomal analysis on microfluidic platforms. IET Nanobiotechnol 1(3):27–35

    Article  Google Scholar 

  • Sieben VJ, Debes-Marun CS, Pilarski LM, Backhouse CJ (2008) An integrated microfluidic chip for chromosome enumeration using fluorescence in situ hybridization. Lab Chip 8:2151–2156

    Article  Google Scholar 

  • Southern E, Mir K, Shchepinov M (1999) Molecular interactions on microarrays. Nat Genetics 21:5–9

    Article  Google Scholar 

  • Vanneste E, Melotte C, Debrock S, D’Hooghe T, Brems H, Fryns JP, Legius E, Vermeesch JR (2009) Preimplantation genetic diagnosis using fluorescent in situ hybridization for cancer predisposition syndromes caused by microdeletions. Hum Reprod 24:1522–15238

    Article  Google Scholar 

  • Wang L, Li PCH (2010) Optimization of a microfluidic microarray device for the fast discrimination of fungal pathogenic DNA. Anal Biochem 400:282–288

    Article  Google Scholar 

  • Weise A, Liehr T, Claussen U, Halbhuber K (2005) Increased efficiency of fluorescence in situ hybridization (FISH) using the microwave. J Histochem Cytochem 53:1301–1303

    Article  Google Scholar 

  • Weng CH, Lien KY, Yang SY, Lee GB (2011) A suction-type, pneumatic microfluidic device for liquid transport and mixing. Microfluid Nanofluid 10:301–310

    Article  Google Scholar 

  • Yang YN, Hsiung SK, Lee GB (2009) A pneumatic micropump incorporated with a normally closed valve capable of generating a high pumping rate and a high back pressure. Microfluid Nanofluid 6:823–833

    Article  Google Scholar 

  • Ye CJ, Stevens JB, Liu G, Ye KJ, Yang F, Bremer SW, Heng HHQ (2006) Combined multicolor-FISH and immunostaining. Cytogenet Genome Res 114:227–234

    Article  Google Scholar 

  • Yuen PK, Li G, Bao Y, Müller UR (2003) Microfluidic devices for fluidic circulation and mixing improve hybridization signal intensity on DNA arrays. Lab Chip 3:46–50

    Article  Google Scholar 

Download references

Acknowledgments

Authors would like to thank the National Science Council, Taiwan, for financial support (NSC 101-2120-M-007-014). Partial financial support from the “Towards A World-class University” Project is also greatly appreciated.

Author information

Authors and Affiliations

  1. Department of Engineering Science, National Cheng Kung University, Tainan, 70101, Taiwan

    Chien-Hsuan Tai

  2. Institute of Basic Medical Science, College of Medicine, National Cheng Kung University, Tainan, 70101, Taiwan

    Chung-Liang Ho & Ya-Lan Chen

  3. Laboratory of Molecular Diagnostics, Department of Pathology, National Cheng Kung University Hospital, Tainan, 70403, Taiwan

    Chung-Liang Ho & Wan Li Chen

  4. Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, 30013, Taiwan

    Gwo-Bin Lee

Authors
  1. Chien-Hsuan Tai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chung-Liang Ho
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ya-Lan Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wan Li Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gwo-Bin Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Chung-Liang Ho or Gwo-Bin Lee.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tai, CH., Ho, CL., Chen, YL. et al. A novel integrated microfluidic platform to perform fluorescence in situ hybridization for chromosomal analysis. Microfluid Nanofluid 15, 745–752 (2013). https://doi.org/10.1007/s10404-013-1190-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10404-013-1190-0

Keywords

Search

Navigation