# Deep learning-enabled virtual multiplexed immunostaining of label-free tissue for vascular invasion assessment

arXiv: https://arxiv.org/abs/2508.16209 · HTML: https://mlsanity.com/paper/2508.16209/

## Key facts

- Cited by 1 papers in the map (data through 14 Oct 2026).
- Builds on 24 of its 63 references that are in the map.
- First posted on arXiv 22 Aug 2025.

## Builds on (most cited first, 24 of 24)

- [Deep Residual Learning for Image Recognition](https://mlsanity.com/paper/1512.03385/) (2015)
- [Denoising Diffusion Probabilistic Models](https://mlsanity.com/paper/2006.11239/) (2020)
- [PyTorch: An Imperative Style, High-Performance Deep Learning Library](https://mlsanity.com/paper/1912.01703/) (2019)
- [The Unreasonable Effectiveness of Deep Features as a Perceptual Metric](https://mlsanity.com/paper/1801.03924/) (2018)
- [Denoising Diffusion Implicit Models](https://mlsanity.com/paper/2010.02502/) (2020)
- [Generative Adversarial Networks](https://mlsanity.com/paper/2203.00667/) (2022)
- [Image-to-Image Translation with Conditional Adversarial Networks](https://mlsanity.com/paper/1611.07004/) (2016)
- [Attention U-Net: Learning Where to Look for the Pancreas](https://mlsanity.com/paper/1804.03999/) (2018)
- [Visual Autoregressive Modeling: Scalable Image Generation via Next-Scale Prediction](https://mlsanity.com/paper/2404.02905/) (2024)
- [VoxelMorph: A Learning Framework for Deformable Medical Image Registration](https://mlsanity.com/paper/1809.05231/) (2018)
- [Breaking the Dilemma of Medical Image-to-image Translation](https://mlsanity.com/paper/2110.06465/) (2021)
- [Deep learning-based virtual histology staining using auto-fluorescence of label-free tissue](https://mlsanity.com/paper/1803.11293/) (2018)
- [Deep Learning-enabled Virtual Histological Staining of Biological Samples](https://mlsanity.com/paper/2211.06822/) (2022)
- [PhaseStain: Digital staining of label-free quantitative phase microscopy images using deep learning](https://mlsanity.com/paper/1807.07701/) (2018)
- [Digital synthesis of histological stains using micro-structured and multiplexed virtual staining of label-free tissue](https://mlsanity.com/paper/2001.07267/) (2020)
- [Label-free virtual HER2 immunohistochemical staining of breast tissue using deep learning](https://mlsanity.com/paper/2112.05240/) (2021)
- [Virtual staining of defocused autofluorescence images of unlabeled tissue using deep neural networks](https://mlsanity.com/paper/2207.02946/) (2022)
- [Virtual histological staining of unlabeled autopsy tissue](https://mlsanity.com/paper/2308.00920/) (2023)
- [Virtual Histological Staining of Label-Free Total Absorption Photoacoustic Remote Sensing (TA-PARS)](https://mlsanity.com/paper/2203.02584/) (2022)
- [Virtual birefringence imaging and histological staining of amyloid deposits in label-free tissue using autofluorescence microscopy and deep learning](https://mlsanity.com/paper/2403.09100/) (2024)
- [Pixel super-resolved virtual staining of label-free tissue using diffusion models](https://mlsanity.com/paper/2410.20073/) (2024)
- [A robust and scalable framework for hallucination detection in virtual tissue staining and digital pathology](https://mlsanity.com/paper/2404.18458/) (2024)
- [Virtual Staining of Label-Free Tissue in Imaging Mass Spectrometry](https://mlsanity.com/paper/2411.13120/) (2024)
- [Label-free evaluation of lung and heart transplant biopsies using tissue autofluorescence-based virtual staining](https://mlsanity.com/paper/2409.05255/) (2024)

## Built on by (most cited first, 1 of 1)

- [Autonomous labeling of surgical resection margins using a foundation model](https://mlsanity.com/paper/2511.22131/) (2025)

## Abstract (arXiv)

Immunohistochemistry (IHC) has transformed clinical pathology by enabling the visualization of specific proteins within tissue sections. However, traditional IHC requires one tissue section per stain, exhibits section-to-section variability, and incurs high costs and laborious staining procedures. While multiplexed IHC (mIHC) techniques enable simultaneous staining with multiple antibodies on a single slide, they are more tedious to perform and are currently unavailable in routine pathology laboratories. Here, we present a deep learning-based virtual multiplexed immunostaining framework to simultaneously generate ERG and PanCK, in addition to H&E virtual staining, enabling accurate localization and interpretation of vascular invasion in thyroid cancers. This virtual mIHC technique is based on the autofluorescence microscopy images of label-free tissue sections, and its output images closely match the histochemical staining counterparts (ERG, PanCK and H&E) of the same tissue sections. Blind evaluation by board-certified pathologists demonstrated that virtual mIHC staining achieved high concordance with the histochemical staining results, accurately highlighting epithelial cells and endothelial cells. Virtual mIHC conducted on the same tissue section also allowed the identification and localization of small vessel invasion. This multiplexed virtual IHC approach can significantly improve diagnostic accuracy and efficiency in the histopathological evaluation of vascular invasion, potentially eliminating the need for traditional staining protocols and mitigating issues related to tissue loss and heterogeneity.