Chan Ronald C K, Li Joshua J X, Yeung W, Chan Anthony W H
Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong.
Diagn Cytopathol. 2020 May;48(5):417-423. doi: 10.1002/dc.24344. Epub 2020 Feb 4.
With the continuous development of new antibodies, the use of immunohistochemistry (IHC) is becoming more often a requirement. IHC is frequently necessary for establishing cancer diagnosis and making therapeutic decisions. However, cytology specimens such as effusion fluid and fine-needle aspiration are highly variable in cellularity and contain background inflammatory and mesothelial cells. Compared to biopsy or excision specimens, tissue exhaustion and levels of sections not matching are more commonly encountered. We present a method of integrating whole-slide cell block image of multiple antibodies by digital image processing and reconstructing a virtual multiplex IHC image for enhanced interpretation.
Historic archived cell block preparations of carcinomas (n = 19) and melanoma (n = 1) and IHC performed with 3,3'-diaminobenzidine chromogen were reviewed. The slides were digitized by a whole-slide image scanner. Using ImageJ and self-developed code, the slides were aligned by image registration, layered, and recolored to reconstruct a virtual multiplex image, simulating a multiplex preparation with multiple chromogens. To quantity the performance of the image registration, the mean distance between the same cell clusters in aligned images were measured.
All 20 cases were successfully registered. The mean distance between cell clusters after image registration was 8.40 +/- 5.52 μm. The reconstructed images were able to demonstrate coexpression of membranous, cytoplasmic, and nuclear antibodies.
With virtual multiplex IHC, we were able to visualize coexpression of multiple antibodies without the added cost of multiplex IHC. Routine and historic slides, without additional tissue consumption, can be retrieved for image reconstruction. This technique is a low-cost adjunct to diagnosis in cytology for more efficient and accurate assessment of antibody coexpression and histochemical stains.
随着新型抗体的不断发展，免疫组织化学（IHC）的应用越来越频繁。IHC对于癌症诊断和治疗决策通常是必要的。然而，诸如积液和细针穿刺等细胞学标本的细胞数量差异很大，并且含有背景炎症细胞和间皮细胞。与活检或切除标本相比，组织耗尽和切片不匹配的情况更常见。我们提出了一种通过数字图像处理整合多种抗体的全玻片细胞块图像并重建虚拟多重免疫组化图像以增强解读的方法。
回顾了癌（n = 19）和黑色素瘤（n = 1）的历史存档细胞块制剂以及使用3,3'-二氨基联苯胺显色剂进行的免疫组化。通过全玻片图像扫描仪对玻片进行数字化处理。使用ImageJ和自行开发的代码，通过图像配准对玻片进行对齐、分层和重新着色，以重建虚拟多重图像，模拟使用多种显色剂的多重制剂。为了量化图像配准的性能，测量了对齐图像中相同细胞簇之间的平均距离。
所有20例均成功配准。图像配准后细胞簇之间的平均距离为8.40 +/- 5.52μm。重建图像能够显示膜、细胞质和核抗体的共表达。
通过虚拟多重免疫组化，我们能够在不增加多重免疫组化成本的情况下可视化多种抗体的共表达。无需额外消耗组织即可检索常规和历史玻片用于图像重建。这项技术是细胞学诊断的低成本辅助手段，可更高效、准确地评估抗体共表达和组织化学染色。
