Application of a microplate scale fluorochrome staining assay for the assessment of viability of probiotic preparations

Hanna-Leena Alakomi (Corresponding Author), Jaana Mättö, Ilkka Virkajärvi, Maria Saarela

Research output: Contribution to journalArticleScientificpeer-review

39 Citations (Scopus)

Abstract

Cell viability in probiotic preparations is traditionally assessed by the plate count technique. Additionally, fluorescent staining combined with epifluorescence microscopy or flow cytometry has been developed for the viability assessment, but the currently available assays are either laborious or require highly sophisticated equipment. The aim of this study was to investigate the applicability of a microplate scale fluorochrome assay for predicting the cell state of freeze-dried Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis preparations. In addition to viability assessment with LIVE/DEAD® BacLight™ Bacterial Viability Kit, DiBAC4(3) stain was used for the kinetic measurement of changes in bifidobacterial cell membrane functions during exposure to low pH. The microplate scale fluorochrome assay results on the viability and cell numbers of probiotic preparations correlated well with the results obtained with the culture-based technique and (with few exceptions) with epifluorescence microscopy. The assay was applicable also for the viability assessment of stressed (acid-treated) cells provided that the cell density in treatments was adjusted to the optimal measurement level of the fluorometer. The microplate scale fluorochrome assay offers a rapid and robust tool for the viability assessment of probiotic preparations, and enables also kinetic measurements.
Original languageEnglish
Pages (from-to)25 - 35
Number of pages11
JournalJournal of Microbiological Methods
Volume62
Issue number1
DOIs
Publication statusPublished - 2005
MoE publication typeA1 Journal article-refereed

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Probiotics
Fluorescent Dyes
Staining and Labeling
Microscopy
Cell Count
Lactobacillus rhamnosus
Microbial Viability
Culture Techniques
Cell Survival
Flow Cytometry
Coloring Agents
Cell Membrane
Equipment and Supplies
Acids

Keywords

  • fluorescent stain
  • fluorometry
  • viability
  • Lactobacillus
  • Bifidobacterium

Cite this

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abstract = "Cell viability in probiotic preparations is traditionally assessed by the plate count technique. Additionally, fluorescent staining combined with epifluorescence microscopy or flow cytometry has been developed for the viability assessment, but the currently available assays are either laborious or require highly sophisticated equipment. The aim of this study was to investigate the applicability of a microplate scale fluorochrome assay for predicting the cell state of freeze-dried Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis preparations. In addition to viability assessment with LIVE/DEAD{\circledR} BacLight™ Bacterial Viability Kit, DiBAC4(3) stain was used for the kinetic measurement of changes in bifidobacterial cell membrane functions during exposure to low pH. The microplate scale fluorochrome assay results on the viability and cell numbers of probiotic preparations correlated well with the results obtained with the culture-based technique and (with few exceptions) with epifluorescence microscopy. The assay was applicable also for the viability assessment of stressed (acid-treated) cells provided that the cell density in treatments was adjusted to the optimal measurement level of the fluorometer. The microplate scale fluorochrome assay offers a rapid and robust tool for the viability assessment of probiotic preparations, and enables also kinetic measurements.",
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Application of a microplate scale fluorochrome staining assay for the assessment of viability of probiotic preparations. / Alakomi, Hanna-Leena (Corresponding Author); Mättö, Jaana; Virkajärvi, Ilkka; Saarela, Maria.

In: Journal of Microbiological Methods, Vol. 62, No. 1, 2005, p. 25 - 35.

Research output: Contribution to journalArticleScientificpeer-review

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AU - Mättö, Jaana

AU - Virkajärvi, Ilkka

AU - Saarela, Maria

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