Survival of BSC-1 cells through the maintenance of cell volume brought about by epidermal growth factor depends on attachment to the substratum

Shiba, Yoshiki; Kanno, Yoshinobu

doi:10.1007/bf01954243

Cited by 2 publications

(2 citation statements)

References 17 publications

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“…The majority of cell volume studies have been performed on cell suspensions using the Coulter counting technique or light scatter; the volume of detached cells can also be measured under a regular light microscope if the cells are assumed to be spherical [24,45]. By contrast, volume measurements on adherent cells are far from straightforward.…”

Section: Introductionmentioning

confidence: 99%

Measurement of the thickness and volume of adherent cells using transmission-through-dye microscopy

Gregg

McGuire

Focht³

et al. 2010

Pflugers Arch - Eur J Physiol

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Cell volume is one of the basic characteristics of a cell and is being extensively studied in relationship to a variety of processes, such as proliferation, apoptosis, fertility, or locomotion. At the same time, its measurement under a microscope has not been well developed. The method we propose uses negative transmission contrast rendered to cells by a strongly absorbing dye present in the extracellular medium. Cells are placed in a shallow compartment, and a nontoxic and cell-impermeant dye, such as acid blue 9, is added to the medium. Transmission images are collected at the wavelength of maximum dye absorption (630 nm). Where the cell body displaces the dye, the thickness of the absorbing layer is reduced; thus, an increase in cell thickness produces brighter images and vice versa. The absolute values for cell thickness and volume can be easily extracted from the image by computing the logarithm of intensity and dividing it by the absorption coefficient. The method is fast, impervious to instability of the light source, and has a high signal-to-noise ratio; it can be realized either on a laser scanning or a conventional microscope equipped with a bandpass filter. For long-term experiments, we use a Bioptechs perfusion chamber fitted with a 0.03-mm spacer and an additional port to enable rapid switching of solutions. To show possible applications of this method, we investigated the kinetics of the cell volume response to a hypotonic buffer and to the apoptotic agents staurosporine and ionomycin.

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Section: Introductionmentioning

confidence: 99%

Measurement of the thickness and volume of adherent cells using transmission-through-dye microscopy

Gregg

McGuire

Focht³

et al. 2010

Pflugers Arch - Eur J Physiol

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“…Cells are assumed to be approximately spherical; their diameters d or cross‐sectional areas are estimated and used to calculate the volume as ⅙πd 3 . Imaging is usually done in transmitted light but fluorescence can also be used . It is obvious that representing even a loosely attached cell with a sphere (rather than a spheroid or some other shape) requires a certain leap of faith from the experimenter, and direct testing of cell sphericity is rarely done .…”

Section: Geometrical Approachesmentioning

confidence: 99%

Methods for cell volume measurement

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2017

Cytometry Pt A

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Volume is an essential characteristic of a cell, and this review describes the main methods of its measurement that have been used in the past several decades. The discussed methods include various implementations of light scattering, estimates based on one or two cell dimensions, surface scanning, fluorescence confocal and transmission slice-by-slice imaging, intracellular volume markers, displacement of extracellular solution, quantitative phase imaging, radioactive methods, and some others. Suitability of these methods to some typical samples and applications is discussed. © 2017 International Society for Advancement of Cytometry.

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Survival of BSC-1 cells through the maintenance of cell volume brought about by epidermal growth factor depends on attachment to the substratum

Cited by 2 publications

References 17 publications

Measurement of the thickness and volume of adherent cells using transmission-through-dye microscopy

Measurement of the thickness and volume of adherent cells using transmission-through-dye microscopy

Methods for cell volume measurement

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