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Improvement of Bacillus subtilis Spore Enumeration and Label Analysis in Flow Cytometry
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Biochemistry
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JoVE Journal Biochemistry
Improvement of Bacillus subtilis Spore Enumeration and Label Analysis in Flow Cytometry

Improvement of Bacillus subtilis Spore Enumeration and Label Analysis in Flow Cytometry

DOI:
10.3791/65141-v

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06:39 min

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June 30, 2023

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, , , , , , , , , , , ,
1Laboratório de diagnóstico e controle de doenças infecciosas na Amazônia,Instituto Leônidas e Maria Deane – Fiocruz Amazônia, 2Programa de Pós-Graduação em Biotecnologia, Instituto de Ciências Biológicas,Universidade Federal do Amazonas (UFAM), 3Programa de Pós-Graduação em Biologia da Interação Patógeno-Hospedeiro – Fiocruz Amazônia, 4Programa de Pós-graduação em Ciências Aplicadas à Hematologia PPGH-UEA/HEMOAM, 5Department of Biology,Federico II University, 6Programa de Pós-Graduação em Imunologia Básica e Aplicada, Instituto de Ciências Biológicas,Universidade Federal do Amazonas (UFAM), 7Laboratório de Imunologia Celular Aplicada à saúde, Fundação Oswaldo Cruz,FIOCRUZ Rondônia, 8Centro Multiusuário para Análises de Fenômenos Biomédicos (CMABio-UEA),Universidade Estadual do Amazonas, 9Universidade Federal do Amazonas (UFAM)

Chapters

  • 00:00Introduction
  • 00:44Flow Cytometry Setting and Preparation of Autoclaved Spores for Flow Cytometry
  • 01:47Analysis Using Flow Cytometry
  • 03:27Estimation of the Protein Coupling Index on a Spore Surface Using Flow Cytometry
  • 04:52Results: Fluorescence Coupling Percentage Analysis of Bacillus subtilis Spores
  • 06:08Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol focuses on the use of flow cytometry and counting beads to quantify bacterial spores labeled with ethidium bromide. The method is also efficient for analyzing the covalent coupling of proteins on the surface of intact spores.

Tags

Bacillus SubtilisSpore EnumerationFlow CytometryCell CounterDormant SporesGerminating SporesFluorescent ProteinsEthidium BromidePBSBeadsGating StrategyLaser PowerThresholdAutofluorescenceCompensationAcquisitionEDACN-hydroxysuccinimide

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