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Rapid micro-immunohistochemistry
Robert D Lovchik1, David Taylor1,2, Govind Kaigala1
1IBM Research Europe, Saeumerstrasse 4, 8803 Rueschlikon, Switzerland.
Insights
We developed rapid immunohistochemistry (IHC) using microfluidic probes, reducing staining times to under 30 minutes. This innovation enables faster protein detection in tissues, potentially aiding surgical decisions and multiplexed assays.
Area of Science:
- Biomedical Engineering
- Histopathology
- Biochemistry
Background:
- Immunohistochemistry (IHC) is crucial for detecting specific proteins in tissue sections.
- Standard IHC protocols involve lengthy incubation steps, limiting rapid analysis.
- There is a need for faster, more efficient IHC methods, especially for intraoperative applications.
Purpose of the Study:
- To develop and validate a rapid and localized immunohistochemical staining method.
- To significantly reduce the incubation times of key IHC steps.
- To explore the potential of this method for intraoperative analysis and multiplexed assays.
Main Methods:
- Fabrication of horizontally oriented microfluidic probes (MFPs) with designed apertures.
- Application of MFPs for localized and time-optimized exposure of tissue sections to biochemicals.
- Validation of the rapid IHC protocol on MDAMB468-1510A cell block sections.
Main Results:
- Achieved reduction of two main IHC incubation steps to less than 30 minutes.
- Demonstrated successful protein detection with rapid incubation times.
- Showcased conservative tissue usage suitable for multiplexed assays.
Conclusions:
- The developed rapid IHC method offers a versatile and efficient alternative to standard protocols.
- This technique has the potential for real-time application during surgery for dynamic clinical decision-making.
- The method supports multiplexed assays, optimizing conditions with minimal tissue samples.
Abstract:
We present a new and versatile implementation of rapid and localized immunohistochemical staining of tissue sections. Immunohistochemistry (IHC) comprises a sequence of specific biochemical reactions and allows the detection of specific proteins in tissue sections. For the rapid implementation of IHC, we fabricated horizontally oriented microfluidic probes (MFPs) with functionally designed apertures to enable square and circular footprints, which we employ to locally expose a tissue to time-optimized sequences of different biochemicals. We show that the two main incubation steps of IHC protocols can be performed on MDAMB468-1510A cell block sections in less than 30 min, compared to incubation times of an hour or more in standard protocols. IHC analysis on the timescale of tens of minutes could potentially be applied during surgery, enabling clinicians to react in more dynamically and efficiently. Furthermore, this rapid IHC implementation along with conservative tissue usage has strong potential for the implementation of multiplexed assays, allowing the exploration of optimal assay conditions with a small amount of tissue to ensure high-quality staining results for the remainder of the sample.
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