Manufacturing method of blood circulating tumor cell pathological chip

Abstract

The invention relates to a manufacturing method of a blood circulating tumor cell pathological chip. Specifically, the invention relates to liquid-based staining of circulating tumor cells in a blood sample, and detection of the circulating tumor cells in a pathological section. The invention also relates to a pathological chip for carrying out the above method.

Description

technical field [0001] The present invention relates to the detection of blood circulating tumor cells (Circulating Tumor Cells, CTCs), more specifically to the staining of circulating tumor cells in blood samples, and then making them into a pathology chip for morphological diagnosis and biological identification. The present invention also relates to a pathology chip and a kit for performing the above method. Background technique [0002] Conventional cytopathology (cytology) pathological slides are generally prepared by using patient blood and other body fluids (1) to smear directly on the slide, (2) or to fix the cells in the liquid on the slide by centrifugal force ( cytospin), (3) or through the centrifugation process, the cells in the liquid are formed into a sediment (cell block), and then subjected to routine pathological section processing, and then the pathologist observes the cells under a microscope for cytopathological diagnosis. [0003] Circulating tumor cel...

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Problems solved by technology

[0007] 1) The dyes of conventional cytopathological staining methods will affect the binding of the first antibody or the first probe to the relevant target, so these stains need to be washed to better realize the immunohistochemical staining of biomarkers

2) After cleaning, a higher concentration of protein antibody or nucleic acid probe is often required for staining than conventional staining, which is likely to cause false positives and mislead clinical diagnosis, treatment and prognosis

4) Even if routine staining does not need to be removed in individual cases, the process of immunohistochemical staining or chromogenic in situ hybridization staining can lead to the loss of conventional dyes, resulting in poor observation of cell morphology under conventional staining

5) The usual immunohistochemistry or nucleic acid chromogenic in situ hybridization staining, in order to improve the sensitivity of the staining, the process of chemical staining will be improved as much as possible, which will cause excessive accumulation of chromogenic products and cover the cells, affecting the diagnosis of observation morphology

However, in this method, the cells are first fixed on the glass slide, and the slide is washed several times in an open system before staining. In this way, the cells will be lost during the staining process, which makes the rare CTC cells become fewer

In addition, the patent application uses immunofluorescence staining, which requires a fluorescent microscope and other special scanning equipment, and cannot provide morphological characteristics of cells that can be used for pathological diagnosis

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