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Direct immunohistochemistry and immunocytochemistry methods

A cell and antibody technology, applied in the field of direct immunohistochemistry and immunocytochemistry, which can solve problems such as time increase

Inactive Publication Date: 2021-01-15
贵州美鑫达医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, many IHC techniques can take 60 to 120 minutes to obtain results
For example, the time required to perform IHC techniques may increase when more than one target antigen is assessed and / or chemical stains are also applied to the sample

Method used

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  • Direct immunohistochemistry and immunocytochemistry methods
  • Direct immunohistochemistry and immunocytochemistry methods
  • Direct immunohistochemistry and immunocytochemistry methods

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0310] This example shows double IHC staining of frozen skin tissue to distinguish cancerous from healthy tissue.

[0311] Direct IHC using an anti-CK5 HRP-polymerized enzyme / antibody conjugate (polyHRP-CK5) and an anti-CK8 / 18 HRP-polymerized enzyme / antibody conjugate (polyHRP-CK8 / 18) was performed on intraoperatively obtained frozen skin tissue sections dyeing. CK5 is a cytokeratin that has demonstrated utility for determining surgical margins during skin cancer surgery when targeting basal and squamous cell carcinomas. However, when only CK5 is targeted during IHC staining, the morphology of eccrine gland cells (such as sweat glands or sebaceous glands) may make it difficult to distinguish healthy from cancerous tissue. To more accurately determine margins for surgical treatment of skin cancer, double staining with poly-HRP-CK5 and poly-HRP-CK8 / 18 was performed on frozen surgical margins from patients with squamous carcinoma. HRP was inactivated by 30% H2O2 for 30 minutes ...

Embodiment 2

[0314] This example shows triple staining of formalin-fixed paraffin-embedded (FFPE) breast cancer tissue.

[0315] Anti-HER2 HRP polymerized enzyme / antibody conjugates (polyHRP-Herceptin (polyHRP-Herceptin)), anti-CK8 / 18 HRP polymerized enzyme / antibody conjugates (polyHRP-CK8 / 18), and anti-Ki -67 HRP polymerized enzyme / antibody conjugate (polyHRP-Ki-67) performed direct IHC staining.

[0316] Breast cancer tissue was first stained with poly-HRP-Herceptin using DAB as the chromogen to generate a brown color in the membrane of the cancer cells. Following inactivation of HRP by 30% H2O2 for 30 minutes, the tissue was then stained with poly-HRP-CK8 / 18 and yellow chromogen to stain the cytoplasm. After a second inactivation step to inactivate HRP, the tissue was further stained with poly-HRP-ki67 and AEC as a chromogen, which produces a red-purple color in the nucleus.

[0317] Improved images of stained tissues were obtained using a triple staining approach in which poly-HRP-He...

Embodiment 3

[0319] This example shows direct ICC staining of cell smears from fine needle aspiration biopsies of the lung. Fine-needle aspiration biopsy samples were previously stained with hematoxylin and eosin and had been archived for many years.

[0320] Direct ICC staining was performed using an anti-CK8 / 18 HRP polymerized enzyme / antibody conjugate (polyHRP-CK8 / 18). Poly-HRP-CK8 / 18 stained the cytoplasm of intact epithelial cells from cytological samples previously stained with the chemical stains hematoxylin and eosin ( Figure 4 ).

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Abstract

The present disclosure provides methods, kits, and compositions for direct immunohistochemical (IHC) staining and direct immunocytochemistiy (ICC) techniques, including applications to multiplex assays, chemically stained samples, and cytology samples.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of priority to U.S. Provisional Application No. 62 / 628,723, filed February 9, 2018, which is hereby incorporated by reference in its entirety. technical field [0003] The present disclosure provides methods, kits, and compositions for direct immunohistochemical (IHC) staining and direct immunocytochemical (ICC) techniques, including applications to multiplex assays, chemically stained samples, and cytological samples. Background technique [0004] Immunohistochemical (IHC) staining of heterologous tissue samples is a reliable method for assessing the presence or absence of target epitopes of target antigens, such as proteins. Typically, IHC techniques utilize antibodies to detect and visualize cellular antigens, eg, target epitopes, in situ. However, since target epitopes are often diffusely distributed in tissue and cellular samples, signal amplification is necessary to visualize ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/547C07K16/46C07K19/00
CPCG01N33/543C07K16/46G01N33/547C07K19/00C07K2319/33G01N33/535G01N2333/916G01N2474/20C07K16/18G01N33/581G01N33/5088G01N33/57415C12N9/00G01N33/537G01N33/533C12N9/99G01N1/30G01N2001/302
Inventor S·陈J·V·吴J·王
Owner 贵州美鑫达医疗科技有限公司
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