Measurement of bacterial volume by transmission-through-dye imaging

Lababidi, Suzanne; Pelts, Mariana; Moitra, Moumita; Leff, Laura Gunn; Model, Michael A.

doi:10.1016/j.mimet.2011.08.019

Cited by 8 publications

(6 citation statements)

References 17 publications

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“…Previously TTD microscopy was used for mammalian cells or pure bacterial cultures [8]. Here we present several examples that show that TTD is highly effective for recognizing live cells in environmental samples, even those heavily contaminated with dead matter and detritus.…”

Section: Discussionmentioning

confidence: 83%

Observation of Living Organisms in Environmental Samples by Transmission-Through-Dye Microscopy

Model

Davis

2016

Micros. Today

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Transmission-through-dye (TTD) microscopy renders live cells bright red over a dark background. Previously TTD had been used for cell volume measurements; in this article the strong contrast generated by TTD is presented as a stand-alone merit, in particular for observation of environmental samples. Such samples often contain large amounts of detritus that can easily obscure living cells and organisms. TTD imaging allows the microscopist to circumvent this difficulty: cells with intact membranes turn bright on TTD images, whereas dead matter remains dark. This property of TTD can be valuable for observing and studying aquatic life.

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Section: Discussionmentioning

confidence: 83%

Observation of Living Organisms in Environmental Samples by Transmission-Through-Dye Microscopy

Model

Davis

2016

Micros. Today

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“…To confirm that the Surface-FISH signals are not a result of RNA leakage from damaged cell membranes, we combined Malat1 Surface-FISH with a transmission-through-dye (TTD) microscopic analysis, where only live cells with intact membranes are fluorescently labeled [ 16 – 18 ] (Additional file 1 : Fig. S3).…”

Section: Resultsmentioning

confidence: 99%

Natural display of nuclear-encoded RNA on the cell surface and its impact on cell interaction

et al. 2020

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Background Compared to proteins, glycans, and lipids, much less is known about RNAs on the cell surface. We develop a series of technologies to test for any nuclear-encoded RNAs that are stably attached to the cell surface and exposed to the extracellular space, hereafter called membrane-associated extracellular RNAs (maxRNAs). Results We develop a technique called Surface-seq to selectively sequence maxRNAs and validate two Surface-seq identified maxRNAs by RNA fluorescence in situ hybridization. To test for cell-type specificity of maxRNA, we use antisense oligos to hybridize to single-stranded transcripts exposed on the surface of human peripheral blood mononuclear cells (PBMCs). Combining this strategy with imaging flow cytometry, single-cell RNA sequencing, and maxRNA sequencing, we identify monocytes as the major type of maxRNA+ PBMCs and prioritize 11 candidate maxRNAs for functional tests. Extracellular application of antisense oligos of FNDC3B and CTSS transcripts inhibits monocyte adhesion to vascular endothelial cells. Conclusions Collectively, these data highlight maxRNAs as functional components of the cell surface, suggesting an expanded role for RNA in cell-cell and cell-environment interactions.

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“…In one method, the V in value is considered as a constant of 2.3 μL·mg −1 based on the general knowledge that all living organisms contain about 70 wt.% water (Bennett et al, 2008). The optical methods concern cell morphology and determine the whole cell volume based on empirical equations (Lababidi et al, 2011;Loferer-Krossbacher et al, 1998). We estimated the cell volume according to the equation W = 435 × V in 0.86 (Loferer-Krossbacher et al, 1998), and obtained a V in value of 3.6 μL·mg −1 , which is 89% higher than the V in value determined by our method (Table 1).…”

Section: Comparison the V In Values Determined By Different Methodsmentioning

confidence: 99%

An accurate method for estimation of the intracellular aqueous volume of Escherichia coli cells

Wang¹,

Zhou²,

Ji³

et al. 2013

Journal of Microbiological Methods

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Measurement of bacterial volume by transmission-through-dye imaging

Cited by 8 publications

References 17 publications

Observation of Living Organisms in Environmental Samples by Transmission-Through-Dye Microscopy

Observation of Living Organisms in Environmental Samples by Transmission-Through-Dye Microscopy

Natural display of nuclear-encoded RNA on the cell surface and its impact on cell interaction

An accurate method for estimation of the intracellular aqueous volume of Escherichia coli cells

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