Microfluidic-based immunohistochemistry for breast cancer diagnosis: a comparative clinical study

  • Fabio Aimi
  • Maria-Giuseppina Procopio
  • Maria Teresa Alvarez Flores
  • Jean-Philippe Brouland
  • Nathalie Piazzon
  • Saška BrajkovicEmail author
  • Diego Gabriel Dupouy
  • Martin Gijs
  • Laurence de Leval
Original Article
Part of the following topical collections:
  1. Quality in Pathology


Breast cancer is a highly heterogeneous disease. The efficacy of tailored therapeutic strategies relies on the precise detection of diagnostic biomarkers by immunohistochemistry (IHC). Therefore, considering the increasing incidence of breast cancer cases, a concomitantly time-efficient and accurate diagnosis is clinically highly relevant. Microfluidics is a promising innovative technology in the field of tissue diagnostic, enabling for rapid, reliable, and automated immunostaining. We previously reported the microfluidic-based HER2 (human epidermal growth factor receptor 2) detection in breast carcinomas to greatly correlate with the HER2 gene amplification level. Here, we aimed to develop a panel of microfluidic-based IHC protocols for prognostic and therapeutic markers routinely assessed for breast cancer diagnosis, namely HER2, estrogen/progesterone receptor (ER/PR), and Ki67 proliferation factor. The microfluidic IHC protocol for each marker was optimized to reach high staining quality comparable to the standard procedure, while concomitantly shortening the staining time to 16 min—excluding deparaffinization and antigen retrieval step—with a turnaround time reduction up to 7 folds. Comparison of the diagnostic score on 50 formaldehyde-fixed paraffin-embedded breast tumor resections by microfluidic versus standard staining showed high concordance (overall agreement: HER2 94%, ER 95.9%, PR 93.6%, Ki67 93.7%) and strong correlation (ρ coefficient: ER 0.89, PR 0.88, Ki67 0.87; p < 0.0001) for all the analyzed markers. Importantly, HER2 genetic reflex test for all discordant cases confirmed the scores obtained by the microfluidic technique. Overall, the microfluidic-based IHC represents a clinically validated equivalent approach to the standard chromogenic staining for rapid, accurate, and automated breast cancer diagnosis.


Microfluidic tissue processor Immunohistochemistry Breast cancer 



The authors thank Noguet Véronique and Prod’Hom Nicole for performing BenchMark stainings, Cloé Bregnard and Monica Esteves for performing FISH analysis, and Jean-Daniel Roman for the support provided with digital imaging.

Authors’ contributions

S.B., D.G.D., M.G., N.P., and L.d.L designed the study. S.B. and N.P. supervised study execution. F.A. ran the experiments. M.T.A.F. and J.P.B. blindly scored the assessed cases. M.G.P. interpreted the results, F.A. and M.G.P designed the figures, and M.G.P and F.A. wrote the paper. All authors contributed to manuscript writing and revision, and they all approved the submitted version. The authors declare that the article is presently not under consideration for publication in another journal and has not been published previously.

Compliance with ethical standards

Conflict of interest

At the submission of the paper, DGD and SB are employed by Lunaphore Technologies SA, which is commercializing a microfluidic-based staining device. DGD and MAMG have equity interest in Lunaphore Technologies SA. The other authors declare no conflict of interest.

Ethical approval

A total of 50 FFPE breast cancer tissue specimens were retrieved from the Pathology Institute of the University Hospital of Lausanne (CHUV) between January 2016 and April 2017 with the ethical committee approval (protocol number 511-12).

Supplementary material

428_2019_2616_MOESM1_ESM.pdf (984 kb)
ESM 1 (PDF 984 kb)
428_2019_2616_MOESM2_ESM.pdf (147 kb)
ESM 2 (PDF 146 kb)
428_2019_2616_MOESM3_ESM.pdf (144 kb)
ESM 3 (PDF 144 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Fabio Aimi
    • 1
  • Maria-Giuseppina Procopio
    • 2
  • Maria Teresa Alvarez Flores
    • 3
  • Jean-Philippe Brouland
    • 3
  • Nathalie Piazzon
    • 3
  • Saška Brajkovic
    • 4
    Email author
  • Diego Gabriel Dupouy
    • 4
  • Martin Gijs
    • 1
  • Laurence de Leval
    • 3
  1. 1.Laboratory of MicrosystemsEcole Polytechnique Fédérale de LausanneLausanneSwitzerland
  2. 2.Institute of Pathology and Molecular PathologyUniversity Hospital ZurichZurichSwitzerland
  3. 3.Institute of PathologyCentre Hospitalier Universitaire VaudoisLausanneSwitzerland
  4. 4.Lunaphore Technologies SALausanneSwitzerland

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