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Related Experiment Video

Updated: May 19, 2026

Separation and Fractionation of Cell Wall and Cell Membrane Proteins from Mycobacterium tuberculosis for Downstream Protein Analysis
06:14

Separation and Fractionation of Cell Wall and Cell Membrane Proteins from Mycobacterium tuberculosis for Downstream Protein Analysis

Published on: September 26, 2025

Cryopreservation of Mycobacterium tuberculosis complex cells.

Zhiquan Shu1, Kris M Weigel, Scott D Soelberg

  • 1Department of Mechanical Engineering, University of Washington, Seattle, Washington, USA.

Journal of Clinical Microbiology
|August 31, 2012
PubMed
Summary

Slow cooling is crucial for cryopreserving Mycobacterium tuberculosis complex cells, significantly improving recovery rates compared to rapid freezing. Optimal cryopreservation media did not impact viability, suggesting intracellular injury is key.

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System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis

Published on: April 5, 2017

Area of Science:

  • Microbiology
  • Cryobiology
  • Cellular Biology

Background:

  • Long-term preservation of Mycobacterium tuberculosis (M. tuberculosis) is vital for diagnostics, drug development, and research.
  • Standardized cryopreservation protocols are needed for M. tuberculosis complex strains.

Purpose of the Study:

  • To investigate the impact of cooling rates and sample matrices on M. tuberculosis complex cryopreservation.
  • To evaluate microbiological culture and microscopy for assessing cryopreservation efficacy.

Main Methods:

  • Cryopreservation of M. tuberculosis complex models (M. bovis BCG, M. tuberculosis H37Ra) in PBS, Middlebrook 7H9 medium (with/without glycerol), and sputum.
  • Assessing viability using microbiological culture and BacLight LIVE/DEAD staining.
  • Evaluating the effect of slow vs. rapid cooling rates.

Main Results:

  • Slow cooling (degrees C/min) significantly enhanced M. tuberculosis complex recovery compared to rapid cooling (liquid nitrogen immersion).
  • No significant difference in viability was observed among PBS, 7H9, and 7H9 plus glycerol cryopreservation media.
  • BacLight LIVE/DEAD staining results were inconsistent with culture data, suggesting intracellular injury over cell envelope damage.

Conclusions:

  • Cooling rate is the most critical factor for successful M. tuberculosis complex cryopreservation.
  • Suboptimal cryopreservation likely causes intracellular injury, leading to loss of viability.
  • Microbiological culture remains the gold standard for assessing M. tuberculosis complex cryopreservation efficacy.