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

DOI :

10.3791/65141-v

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June 30th, 2023

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471 Views

Késsia Caroline Alves^1,2, Yury Oliveira Chaves^1,3,4, Maria Edilene Almeida^1,4, Maria Gabriella Vasconcelos^1,4, Paulo Afonso Nogueira^1,3,6, Joyce Melo^1,9, Jéssica Marques⁸, Juliana Pavan Zuliani⁷, Charles Nunes Boeno⁷, Mauro Valentino Paloschi⁷, Rachele Isticato⁵, Ezio Ricca⁵, Luís André Mariúba^1,2,3,6

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)

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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.

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