Immunofluorescence staining method for paraffin embedded three-dimensional suspension cell clusters

Abstract

The invention relates to an immunofluorescence staining method of paraffin embedded three-dimensional suspension cell clusters. The method comprises the following steps: pretreating the three-dimensional suspension cell clusters, immersing into paraffin liquid to embed sections, carrying out antigen repair, and carrying out immunofluorescence staining. According to the method provided by the invention, cells in the cell cluster are fully dyed, the result is accurate, the overall protein expression and distribution condition of the suspension cell cluster can be observed, and sample observationcan be carried out only by a common fluorescence microscope.

Description

Technical field [0001] The invention relates to the field of cell biology and related detection, in particular to an immunofluorescence staining method for paraffin-embedded three-dimensional suspended cell clusters. Background technique [0002] Immunofluorescence technique (Immunofluorescence technique) is one of the earliest developed labeling immunological techniques. It is a technology established on the basis of immunology, biochemistry and microscopy technology. This method combines antibody molecules with some tracer substances, and uses antigen-antibody reactions to locate antigenic substances in tissues or cells. [0003] At present, the immunofluorescence staining methods for 3D suspension cell clusters (spheres) mainly include: (1) Centrifugation; the defect is that centrifugation will cause partial cell cluster loss, cell damage, disintegration and deformation, and insufficient staining of cells inside the cell cluster. (2) Staining method with the aid of a staining d...

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Problems solved by technology

[0003] At present, the immunofluorescent staining methods for 3D suspension cell clusters (spheres) mainly include: (1) centrifugation; the disadvantage is that centrifugation will cause some loss of cell clusters, damage to cells, loose deformation, and insufficient staining of cells inside the cell clusters.

(2) Staining method with the help of a staining device; the disadvantage is that the removal of residual liquid is not complete or the cell mass is easy to dry out. One device can only do one set of samples, wasting reagents and antibodies

Moreover, the above methods all need to use expensive laser confocal microscope to observe the staining results, and the staining effect of the cells inside the cell mass cannot be judged.

[0004] If the classic paraffin embedding technique is used directly for immunofluorescence staining after 3D suspension cell mass embedding, there will be problems such as poor staining effect, dark background or no staining, cell mass sections are easy to fall off from the glass slide, etc.

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