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Three-Dimensional Microscopy in Microbiology

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...

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Updated: Apr 16, 2026

Industrialized, Artificial Intelligence-guided Laser Microdissection for Microscaled Proteomic Analysis of the Tumor Microenvironment

Industrialized, Artificial Intelligence-guided Laser Microdissection for Microscaled Proteomic Analysis of the Tumor Microenvironment

Published on: June 3, 2022

Multi-Tiered µDicer Enables Protein-Preserving Microdissection at 10 µm Resolution.

Annatoma Arif¹, Rashmi Kumar², Yumi Kwon²

¹Department of Mechanical Engineering, Stanford University, Stanford, California, USA.

Small (Weinheim an Der Bergstrasse, Germany)

|April 15, 2026

Summary

This summary is machine-generated.

Multi-tiered µDicers mechanically dissect tissues into microtissues for proteomic analysis. This method preserves more proteins than laser capture microdissection (LCM), especially at high spatial resolutions.

Keywords:

hierarchical blade arrays mass spectrometry spatial proteomics tissue microdissection two‐photon polymerization

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Area of Science:

Biotechnology
Proteomics
Tissue Engineering

Background:

Laser capture microdissection (LCM) is crucial for isolating regions of interest (ROIs) in tissues for molecular profiling.
Challenges arise when ROIs approach cellular dimensions (~10 µm), as LCM can cause photothermal damage, compromising protein preservation and quantitative accuracy.

Purpose of the Study:

To introduce multi-tiered µDicers, fabricated by two-photon polymerization, as a novel mechanical method for dissecting uniform microtissues down to 10 µm.
To compare the proteomic yield and quality of microtissues generated by µDicers versus conventional LCM.

Main Methods:

Fabrication of multi-tiered µDicers using two-photon polymerization.
Mechanical dissection of ethanol-fixed human squamous cell carcinoma microtissues using µDicers and LCM.
Proteomic analysis of microtissues using Nanodroplet Processing in One pot for Trace Samples (nanoPOTS) and liquid chromatography-mass spectrometry (LC-MS).

Main Results:

µDicers yielded significantly more peptides and proteins than LCM, particularly at 10-20 µm spatial resolution.
Confocal imaging revealed material loss (catapult-associated cavities) in LCM-generated microtissues, unlike µDicers.
Multi-tiered µDicers enabled reproducible microdissection down to 10 µm while maintaining high protein coverage.

Conclusions:

Multi-tiered µDicers offer a superior mechanical microdissection method compared to LCM for high-resolution spatial proteomic studies.
This technology addresses limitations of LCM, improving protein preservation and quantitative fidelity in microtissues.
µDicers have potential to complement LCM in next-generation spatial proteomic workflows.