Dynamic phase microscopy, a new method to detect viable and killed spores and to estimate the heterogeneity of spore populations

Tychinsky, Vladimir P.; Mulyukin, A. L.; Lisovskii, V. V.; Nikolaev, Yu. A.; Kretushev, A. V.; Vyshenskaya, T. V.; Suzina, N. E.; Дуда, В. И.; Эль-Регистан, Г. И.

doi:10.1016/j.asr.2007.03.090

Cited by 4 publications

(9 citation statements)

References 18 publications

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“…This result is of great importance for our methodology since it illustrates the possibility of calculating MCR using Dn þ À Dn À boundary values obtained from the two phase thickness profiles. Table 1 shows refractivity values presented in [6,8] and obtained in the course of our measurements of mitochondria [14], cyan bacteria [15], spores [16], neurons [4,32] and human cells [18][19][20][21]. Marked changes in the phase thickness and refractivity of nucleoli of tumor cells HCT116 were observed under the influence of actinomycin D that inhibits pre-ribosome transcription and synthesis [20].…”

Section: Measurement Resultsmentioning

confidence: 98%

“…The origin of the refractivity component that was found to be dependent on the functional state, further referred to as the metabolic component of refractivity (MCR), remains mysterious. Later, MCR was found in bacteria spores [16], chloroplasts [17], cyan bacteria, and human cells [18][19][20][21]. The phenomenon, surprising in its universal character, led us to the hypothesis that the common factor might be water [5].…”

Section: Introductionmentioning

confidence: 99%

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The metabolic component of cellular refractivity and its importance for optical cytometry

Tychinsky

2009

Journal of Biophotonics

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Initially, it has been shown that the phase thickness and refractivity (the latter interpreted as the difference of the refractivity indices of an object and surrounding medium) depend on the functional state of mitochondria. The refractivity of various objects decreased in response to energy depletion. This dependence was then demonstrated for other biological objects such as cyanobacteria, chloroplasts and human cells. This general response brought about the hypothesis of a certain "universal" factor that links the variable (or metabolic) component of refractivity with the object's functional state. However, the origin of this phenomenon remains unknown. Our hypothesis is founded on the dependence of polarization of bound water molecules and the activity of metabolic processes. Here, we show the results of measurements of metabolic component of refractivity different bio-objects (mitochondria, chloroplasts, spores, cancer cells) obtained using the Coherent Phase Microscope "Airyscan". Estimations indicated high (up to n approximately = 1.41-1.45) values for the equivalent refractive index of structured water in cells.

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Section: Measurement Resultsmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

The metabolic component of cellular refractivity and its importance for optical cytometry

Tychinsky

2009

Journal of Biophotonics

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“…Coherent phase microscopy provides one of the methods for the non-invasive imaging cells and intracellular organelles and diagnosis of metabolic states in individual cells [23][24][25] and energy-transducing organelles (mitochondria [17,18,21] and chloroplasts [22]). In this section, we describe the use of the CPM method for imaging cells and intracellular organelles.…”

Section: Cpm Visualization Of Cells and Energy-transducing Organellesmentioning

confidence: 99%

“…In this article, we summarize the results of our previous works [7,[17][18][19][20][21][22][23][24][25][26][30][31][32][33][34][35][36][37][38] on visualization of metabolic states of individual cells and energy-transducing organelles by the CPM and DPM methods.…”

Section: Introductionmentioning

confidence: 99%

“…New microscopes and image-processing software allow precise quantitative analysis of cell structures and intracellular dynamics [1][2][3][4][5][6][16][17][18][19][20][21][22][23][24][25][26]. The images of objects can be presented in the form of two-dimensional distribution of some optical parameter (e.g., phase difference [1][2][3][4][5][6]17], polarization, intensities of anti-stokes components [14,15], or spectral components of fluorescent responses [8][9][10][11][12]).…”

Section: Introductionmentioning

confidence: 99%

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Interference Microscopy in Cell Biophysics. 2. Visualization of Individual Cells and Energy-Transducing Organelles

Tychinsky

Тихонов

2010

Cell Biochem Biophys

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The coherent phase microscopy (CPM) provides a convenient and non-invasive tool for imaging cells and intracellular organelles. In this article, we consider the applications of the CPM method to imaging different cells and energy-transducing intracellular organelles (mitochondria and chloroplasts). Experimental data presented below demonstrate that the optical path length difference of the object, which is the basic optical parameter measured by the CPM method, can serve as an indicator of metabolic states of different biological objects at cellular and subcellular levels of structural organization.

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Investigating germination and outgrowth of bacterial spores at several scales

Trunet

Carlin

Coroller

2017

Trends in Food Science & Technology

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Background: Spore-forming bacteria are a major cause of food spoilage and food poisoning. Spores thatresist physical and chemical treatments used in the food industry may germinate and multiply. Spore germination, outgrowth and growth constitute a complex and highly heterogeneous process.Scope and approach: Various techniques and methods can be used to observe the germination, outgrowth and early multiplication process of spore-forming bacteria and/or to quantify the impact of environmental conditions on its progress over time within a spore population. These techniques can be classified by different criteria: (i) the scale of analysis, from populations or cells to molecules, and (ii) the number of analyzed objects (cells) and (iii) the potential of the method to describe and/or quantify the impact of lethal or sub-lethal treatments or environmental conditions. Such treatments are applied to a spore population or a single spore and take into account parameters at the cellular level (growth capacity, morphological properties) to molecular level (proteomics, transcriptomics, spore molecularcomposition).Key findings and conclusion: A better understanding and quantification of the germination, outgrowth and growth process require the implementation of several complementary methods. Methods providing information at single and population levels, as well as at molecular and cellular levels, are essential to assess and control the fate of spore-forming bacteria development in food systems

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Dynamic phase microscopy, a new method to detect viable and killed spores and to estimate the heterogeneity of spore populations

Cited by 4 publications

References 18 publications

The metabolic component of cellular refractivity and its importance for optical cytometry

The metabolic component of cellular refractivity and its importance for optical cytometry

Interference Microscopy in Cell Biophysics. 2. Visualization of Individual Cells and Energy-Transducing Organelles

Investigating germination and outgrowth of bacterial spores at several scales

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