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Tissue Homogenization and Cell Lysis

Tissue Homogenization and Cell Lysis

Tissue homogenization involves disintegrating tissue architecture and lysing cells, and is an early step in isolating and analyzing cellular components. The method used for homogenization depends on the sample type, the amount of sample available, the analyte to be obtained, and the sensitivity of the method. These methods are broadly classified as mechanical and non-mechanical methods.
Mechanical methods of tissue homogenization
These methods rely on applying external physical force to disrupt...

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Enrichment of Extracellular Matrix Proteins from Tissues and Digestion into Peptides for Mass Spectrometry Analysis

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Published on: July 23, 2015

Improving Yields in Multi-analyte Extractions by Utilizing Post-homogenized Tissue Debris.

Ala Petersons^1,2, Joseph Carlson³, William Mathieson¹

¹Integrated Biobank of Luxembourg, Dudelange, Luxembourg.

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society

|April 29, 2023

Summary

Recovering DNA from discarded tissue debris after multi-analyte extraction significantly boosts DNA yield. This method, using proteinase K digestion and alcohol precipitation, enhances DNA recovery without compromising integrity.

Keywords:

AllPrep Puregene QIAamp biospecimen science homogenization optimization simultaneous

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Cryosectioning of Contiguous Regions of a Single Mouse Skeletal Muscle for Gene Expression and Histological Analyses

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Published on: December 12, 2016

Area of Science:

Biochemistry
Molecular Biology
Genomics

Background:

Multi-analyte extractions from tissue yield lower DNA, RNA, and protein compared to single-analyte methods.
The discarded tissue debris after homogenization is often a source of untapped biomolecules.

Purpose of the Study:

To investigate methods for improving DNA yield from post-homogenized tissue debris.
To assess the impact of different extraction techniques on DNA integrity and yield.

Main Methods:

Tissue debris underwent further homogenization and simultaneous extraction.
Proteinase K digestion of debris followed by DNA extraction using silica spin columns or alcohol precipitation.
DNA integrity was assessed using long-range PCR, DNA Integrity Numbers, and electropherogram analysis.

Main Results:

Additional homogenizations yielded no RNA, 13% more DNA (degraded), and 161.7% more protein.
Proteinase K digestion followed by alcohol precipitation yielded an additional 203.9% DNA, while silica columns yielded 27.1% more DNA, without compromising integrity.
Validation in 65 tissue blocks showed average additional DNA yields of 31.6% (silica) and 54.8% (alcohol precipitation).

Conclusions:

Discarded tissue debris can be a valuable source for additional DNA recovery.
Proteinase K digestion combined with alcohol precipitation offers a highly effective method for maximizing DNA yield from tissue remnants.
This approach allows researchers to improve DNA yields from initial multi-analyte extractions by reprocessing homogenized tissue debris.