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Volatilization

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Volatilization gravimetry is an analytical technique that measures the mass lost due to the volatilization of the substance. This technique is used to estimate the amount of volatile material in a sample. To perform this method, heat a known amount of the sample to a high temperature in a crucible or other suitable vessel. The volatile substance in the sample evaporates, and the vapor is completely expelled from the crucible either by heating the sample or bubbling a stream of inert gas through...

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Updated: Apr 30, 2026

Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection

Spatial Profiling of Protein and RNA Expression in Tissue: An Approach to Fine-Tune Virtual Microdissection

Published on: July 6, 2022

A practical protocol of processing mineralized tissue for Visium spatial transcriptomics.

Jixing Miao¹, Mingjie Ma¹, Yuhan Guo²

¹Department of Orthopedics, The First Hospital of China Medical University, Shenyang, Liaoning, 110001, China.

Mechanobiology in Medicine

|December 1, 2025

Summary

This summary is machine-generated.

This study presents an optimized spatial transcriptomics protocol for bone tissue. It uses Morse's solution for rapid decalcification and specialized slides to preserve RNA quality and improve gene detection in mineralized samples.

Keywords:

Bone Decalcification RNA degradation Spatial transcriptomics Visium

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Area of Science:

Biotechnology
Molecular Biology
Genomics

Background:

Spatial transcriptomics of mineralized tissues is challenging due to RNA degradation during decalcification.
Conventional EDTA decalcification protocols are lengthy and compromise RNA integrity.
Improved methods are needed for high-resolution gene expression analysis in bone.

Purpose of the Study:

To develop and validate an optimized protocol for spatial transcriptomics of mineralized tissues using 10x Genomics Visium.
To enhance RNA preservation and transcript capture efficiency in bone and fracture callus samples.
To overcome pre-analytical limitations in analyzing gene expression in skeletal tissues.

Main Methods:

Replaced EDTA decalcification with Morse's solution for rapid processing (<24 hours).
Utilized SCHOTT NEXTERION® Slide H (3-D hydrogel-coated) for enhanced adherence of fragile bone sections.
Applied the protocol to intact and fractured mouse femurs for spatial transcriptomics analysis.

Main Results:

Morse's solution enabled rapid decalcification while preserving RNA quality (favorable DV200 values).
The optimized protocol significantly improved transcript capture efficiency for Visium V2 and Visium HD platforms.
Visium V2 averaged 5639 genes/spot; Visium HD averaged 170 genes/8µm bin.

Conclusions:

The developed protocol effectively overcomes major pre-analytical hurdles in mineralized tissue spatial transcriptomics.
This method enables high-resolution spatial transcriptomic profiling of bone with enhanced data quality.
The optimized workflow facilitates deeper insights into the molecular landscape of skeletal tissues.