Simple and Rapid Method for Separating Lactic Acid Bacteria from Commercially Prepared Yogurt

Nakao, Hidenobu; Kim, Je–Deok

doi:10.2116/analsci.19c008

Cited by 3 publications

(3 citation statements)

References 11 publications

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“…The LAB were separated from the yogurt according to a previously described method. 16 In brief, we added 100 L of a yogurt sample diluted 1:10 in sterile Milli-Q water to a 1.5-mL microcentrifuge tube. The sample was then bulk-sheared by vortexing (AS-ONE, TUBE MIXER TRIO TM-1F, rotation speed 1500 rpm).…”

Section: Recovery Of Lactic Acid Bacteria From Yogurt Samplesmentioning

confidence: 99%

“…Recently, we have reported a useful technique for easily separating LAB from commercially prepared yogurt without the need for any chemical or special equipment. 16 This method relies on the destruction and denaturation of casein micelle aggregates by vortex mixing, and can be used to prepare samples containing only LAB. The LAB recovered by this method could be clearly observed by microscopy, enabling both the bacilli and cocci to be directly detected at a single-cell level.…”

Section: Introductionmentioning

confidence: 99%

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Simple and rapid method for selective enumeration of lactic acid bacteria in commercially prepared yogurt by image analysis and K-means clustering

Nakao

Magariyama

2022

ANAL. SCI.

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We developed a simple and rapid method based on the combination of image analysis and kmeans clustering to selectively enumerate cocci and bacilli from among lactic acid bacteria (LAB) in commercially prepared yogurt. We used our previously reported method for recovering only LAB without non-microbial substances from commercial yogurt, and found that the shape and light intensity of LAB cell images taken by optical microscopy were factors that could distinguish cocci and bacilli, allowing the selective enumeration of LAB. Also, k-means clustering was executed on a dataset of the mean light intensity and aspect ratio of each LAB obtained by image analysis, and each LAB in the image could be automatically assigned to either the cocci or bacilli group. The results obtained by this automated method were in good agreement with those obtained by manually counting the LAB under a microscope, with an overall error within 10%. In addition, this method could provide results within a few hours, which is approximately 1/32 of the time required for the conventional colony counting method.

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Section: Recovery Of Lactic Acid Bacteria From Yogurt Samplesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Simple and rapid method for selective enumeration of lactic acid bacteria in commercially prepared yogurt by image analysis and K-means clustering

Nakao

Magariyama

2022

ANAL. SCI.

Self Cite

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“…Moreover, cells have recently been used in drug discovery, drug development, biomedical research, and biological technology and medical applications. Recently, various types of cell separation methods have been developed 5,6 . Especially, a cell separation method with maintaining cell activity is strongly required for cell transplantation.…”

Section: Introductionmentioning

confidence: 99%

Effective Separation for New Therapeutic Modalities Utilizing Temperature-responsive Chromatography

et al. 2021

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In recent years, drug discovery and therapeutics trends have shifted from a focus on small-molecule compounds to biopharmaceuticals, genes, cell therapy, and regenerative medicine. Therefore, new approaches and technologies must be developed to respond to these changes in medical care. To achieve this, we applied a temperature-responsive separation system to purify a variety of proteins and cells. We developed a temperatureresponsive chromatography technique based on a poly(N-isopropylacrylamide) (PNIPAAm)-grafted stationary phase. This method may be applied to various types of protein and cell separation applications by optimizing the properties of the modified polymers used in this system. Therefore, the developed temperature-responsive HPLC columns and temperature-responsive solid phase extraction (TR-SPE) columns can be an effective separation tool for new therapeutic modalities such as monoclonal antibodies, nucleic acid drugs, and cells.

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Visualization of Gene Reciprocity among Lactic Acid Bacteria in Yogurt by RNase H-Assisted Rolling Circle Amplification-Fluorescence In Situ Hybridization

et al. 2021

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Recently, we developed an in situ mRNA detection method termed RNase H-assisted rolling circle amplification-fluorescence in situ hybridization (RHa-RCA-FISH), which can detect even short mRNA in a bacterial cell. However, because this FISH method is sensitive to the sample condition, it is necessary to find a suitable cell permeabilization and collection protocol. Here, we demonstrate its further applicability for detecting intrinsic mRNA expression using lactic acid bacteria (LAB) as a model consortium. Our results show that this method can visualize functional gene expression in LAB cells and can be used for monitoring the temporal transition of gene expression. In addition, we also confirmed that data obtained from bulk analyses such as RNA-seq or microarray do not always correspond to gene expression in individual cells. RHa-RCA-FISH will be a powerful tool to compensate for insufficient data from metatranscriptome analyses while clarifying the carriers of function in microbial consortia. By extending this technique to capture spatiotemporal microbial gene expression at the single-cell level, it will be able to characterize microbial interactions in phytoplankton–bacteria interactions.

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Simple and Rapid Method for Separating Lactic Acid Bacteria from Commercially Prepared Yogurt

Cited by 3 publications

References 11 publications

Simple and rapid method for selective enumeration of lactic acid bacteria in commercially prepared yogurt by image analysis and K-means clustering

Simple and rapid method for selective enumeration of lactic acid bacteria in commercially prepared yogurt by image analysis and K-means clustering

Effective Separation for New Therapeutic Modalities Utilizing Temperature-responsive Chromatography

Visualization of Gene Reciprocity among Lactic Acid Bacteria in Yogurt by RNase H-Assisted Rolling Circle Amplification-Fluorescence In Situ Hybridization

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