Deparaffinization of tissue by electric field generation and ionization

Inventors

Lim, Victor • Schembri, Carol T. • Mccampbell, Adrienne • Yama-Dang, Nazumi Alice

Assignees

Agilent Technologies Inc

Abstract

Paraffin-embedded tissue, which may be disposed on a solid substrate, is prepared by a dry technique that removes paraffin from tissue without adding any liquid to the tissue, thereby rendering the tissue substantially free of paraffin. The dry technique may entail applying heat energy to the tissue effective to melt the paraffin and thereby render it flowable, and applying an electric field. The electric field is effective to impart electrical charge to the paraffin and to move the paraffin out from the tissue due to electrical charge repulsion or attraction, which may be assisted by moving an electrode utilized to generate the electric field relative to the paraffin. The electric field, or both the electric field and the heat energy, may be applied until the tissue is substantially free of paraffin.

Core Innovation

The invention provides a dry tissue deparaffinization approach that removes paraffin from a paraffin-embedded tissue without adding liquid. The approach melts the paraffin embedded in the tissue by applying heat energy to the tissue and then removes the melted paraffin out of the tissue by applying an electric field effective to move the melted paraffin out of the tissue.

The electric-field removal is implemented by an electric field-generating modality that ionizes paraffin in an ionization region and drives the melted paraffin using electrostatic charge effects. The description includes configurations that produce corona discharge and/or plasma in an ionization region exposed to the paraffin, and electrode arrangements in which a first electrode and a second electrode provide the electrical field.

The dry deparaffinization is further linked to tissue handling and downstream compatibility by providing tissue on a solid substrate and minimizing xylene use. The description highlights compatibility with downstream staining, antigen retrieval, and nucleic-acid workflows including DNA/RNA extraction and PCR/next generation sequencing, while addressing oxidative damage to guanosines from conventional xylene use.

Claims Coverage

The document contains three independent claims, each covering a distinct aspect of the same concept: a method of deparaffinizing tissue, a method of preparing tissue substantially free of paraffin without added liquid, and a tissue preparation device that combines a heating device with an electric-field-generating device. Across the independent claims, the inventive features center on melting paraffin by applied heat and removing the melted paraffin by applying an electric field, including continuation until the tissue is substantially free of paraffin.

Overall claim coverage is directed to a coupled approach where paraffin is melted by applied heat energy and then removed from paraffin-embedded tissue by applying an electric field that moves the melted paraffin out of the tissue, including rendering the tissue substantially free of paraffin without adding liquid. The device claim covers a system that includes both a heating device and an electric field-generating device to implement the heat-melt and electric-field movement functions.

Stated Advantages

Documented Applications