Superresolution at the singularities of phase images

Kretushev, A. V.; Tychinskii, V. P.

doi:10.1070/qe2002v032n01abeh002128

Cited by 11 publications

(8 citation statements)

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“…Rapid analysis of the samples allows to characterize populations and single cells, which is necessary for analysis of heterogeneous material (nonsynchronized cultures, clinical samples, etc). Optical approaches to analysis of living biological objects may be used for cancer diagnosis [14]. Our data suggest that CPM is an adequate approach for selection of antitumor reagents whose mechanism of action is associated with the suppression of gene transcription.…”

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confidence: 92%

Coherent Phase Microscopy, a Novel Approach to Study the Physiological State of the Nucleolus

Tychinsky¹,

Kretushev²,

Klemyashov³

et al. 2005

Dokl Biochem Biophys

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The assessment of metabolism of microobjects is a topical problem of biology. Studies of cells and subcellular structures and their responses to extracellular stress stimuli are of fundamental importance. Important practical applications of such studies include the selection of biologically active substances and the diagnostics of metabolic disturbances. The solution of these problems requires methods that allow quantitative analysis of physiological processes in real time using nonfixed single cells, without utilization of fluorescent dyes or other invasive approaches which may introduce experimental artifacts. The heterogeneity of animal cells and microorganisms stimulates the development of novel methods for the analysis of physiological state of single cells and recording of individual response to changed microenvironment [1]. The method of coherent phase microscopy (CPM) developed by our group [2, 3] is based on representation of a real biological object as a spatially heterogeneous distribution of optical refraction index. The phase images obtained by interference techniques represent two-dimensional distributions of optical path difference (or phase thickness). Therefore, the structures with higher optical density are clearly distinguished in such phase images. Hence, the intracellular structures that have different refraction indices will be present in the static phase images. The local changes in the refraction index may be a result of metabolic processes and can be measured as changes in the phase thickness of intracellular structures [2][3][4][5][6][7][8]. CPM was used to study the metabolic state of single large organelles (isolated mitochondria) [4 − 6], plant cells (bean chloroplasts) [7], cyanobacteria, and spores [3]. A decrease in the phase thickness induced by pharmacologic uncoupling of mitochondrial transmembrane potential of protons or inhibition of electron transport (deenergization) was detected [3 − 7]. These studies showed that it is possible to assess the response of living biological objects to changes in homeostasis by CPM.The purposes of this work were (1) to identify the optically dense structures in the whole (unfractionated) cells by the CPM method and (2) to determine optical parameters of the disturbances of the state of nucleoli induced by suppression of gene transcription.The experiments were performed with the cell lines NIH 3T3 (immortalized murine fibroblasts) and HCT116 (transformed human colon epithelium) cultured in Dulbecco's modified Eagle's medium supplemented with 10% bovine serum, 2 mM glutamine, and gentamicin. The cells were maintained at 37°ë in 5% ëé 2 humidified atmosphere. The cells were seeded on cover slips 24 h before experiments; the monolayer density at the day of experiments was 50-75%. The cells were incubated with pharmacological reagents; the cover slips were placed on a polished silicon substrate and analyzed by CPM. Single cells were first studied using the optical channel of CPM; then, their topograms and phase thickness profiles were measured. Abou...

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confidence: 92%

Coherent Phase Microscopy, a Novel Approach to Study the Physiological State of the Nucleolus

Tychinsky¹,

Kretushev²,

Klemyashov³

et al. 2005

Dokl Biochem Biophys

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“…Extension of the notion of spatial resolution to functional images necessitated revision of the classical criterion and interpretation of the term ''super resolution''. Detailed analysis of the problem of spatial resolution in the CPM images is presented in [12,[29][30][31][32][33]]. An essential feature of CPM images is the possibility to detect wave front dislocations by measuring spatial positions of so-called ''singular lines'', where the light intensity tends to zero, I(x,y) = 0, but distinctly increases a steepness of phase changes (''phase gradient'') [29,34].…”

Section: Spatial Resolution Of Objectsmentioning

confidence: 99%

“…The use of a compensation method for measuring the OPLD values ensures significant enhancement of spatial resolution of CPM images given by a CPM ''Airyscan'' [30][31][32][33][34] as compared to the methods based on the multistep [22][23][24][25][26] or other algorithms [10,11]. This approach implies that the local OPLD values, h(x,y), are determined from a zero value of the AC signal for each period of the reference wave phase modulation.…”

Section: Spatial Resolution Of Objectsmentioning

confidence: 99%

“…Methodological advantage of the method is that determination of phase is not limited by diffraction constrains peculiar to measurements of light intensity distribution. This approach provided the realization of super-resolution of samples [12,[27][28][29][30][31][32][33]. For this trend of interference microscopy, we use a term the ''coherent phase microscopy''.…”

Section: Introductionmentioning

confidence: 99%

“…For this trend of interference microscopy, we use a term the ''coherent phase microscopy''. The realization of this method in the coherent phase microscope ''Airyscan'' [27] allowed us to obtain a high spatial resolution of different objects [12,[30][31][32][33][34].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Interference Microscopy in Cell Biophysics. 1. Principles and Methodological Aspects of Coherent Phase Microscopy

Tychinsky

Тихонов

2010

Cell Biochem Biophys

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The purpose of our tandem publications is to review the applications of the coherent phase microscopy to cell biophysics. In this article, we briefly consider the fundamentals and methodological aspects of the coherent phase microscopy (CPM). One of important advantages of this method is a high sensitivity of CPM images to changes in physical-chemical properties of biological samples. The optical path length difference (OPLD), measured with CPM instruments in different domains of an object, serves as the informative optical parameter used for visualization of individual cells or intracellular organelles. Metabolically dependent variations of CPM images reflect changes in the functional state of biological objects. In the next article, we review results of CPM studies of different cells and intracellular energy-transducing organelles.

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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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Superresolution at the singularities of phase images

Cited by 11 publications

References 1 publication

Coherent Phase Microscopy, a Novel Approach to Study the Physiological State of the Nucleolus

Coherent Phase Microscopy, a Novel Approach to Study the Physiological State of the Nucleolus

Interference Microscopy in Cell Biophysics. 1. Principles and Methodological Aspects of Coherent Phase Microscopy

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

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