Real‐Time In Vivo Cellular‐Level Imaging During Puncture

In the pursuit of precise disease diagnosis, accurate tissue sampling during biopsy is critical. Current CT/ ultrasound-guided biopsies provide macroscopic localization but lack real-time cellular-resolution visualization during puncture, particularly for deep, narrow lumens or small lesions, thereby increasing false-negative and nondiagnostic sampling risks. It's desired to have a puncture with microscopy imaging. Here, we demonstrate an artificial-intelligence-empowered integrative-light-field microendoscopy (AIM) needle. This photonic-mechanically co-engineered probe overcomes dynamic diffraction limits via a closed-loop adaptive optics system integrated in a 25G biopsy needle, enabling diffraction-limited imaging during in vivo puncture. AIM needle resolved characteristic layered microstructures throughout mouse organs (parenchymal/hollow) and pulmonary tumors via synergistic light-field modulation, leveraging K-means and convolutional neural networks to enable in situ pathology-like analysis and tumor/normal tissue discrimination along puncture paths. AIM needle demonstrates dual clinical potential as a complement to macroscopic guidance: potentially providing histology-like feedback without interrupting procedures while enhancing biopsy targeting accuracy through navigation integration, reducing false-negative rates in narrow lumens and microlesions, thereby improving early detection sensitivity.