Fast and easy method of lanthanoid staining for visualization of cellular ultrastructure and spatial arrangement

Ia, Novikov; Subbot, A.M.; Kiryushchenkova, N. P.; Нестерова, Т. В.; Габашвили, А. Н.; Ситников, А. В.; Bursov, A. I.

doi:10.1063/1.4954343

Cited by 3 publications

(3 citation statements)

References 4 publications

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“…Physical, structural and biochemical aspects underlie a high image contrast. Lanthanoids, being the elements with a high atomic number, are known to provide a high density of electron backscattering under the impact of a microscope electron beam, which results in a high brightness on the scans of the biological object loci, saturated with these elements [4,7]. Several mechanisms of capturing and deposition of lanthanoids in biological systems are known, one of them is determined by the ability to substitute calcium in the exchange chains, the second one by the ability to actively bind to phosphate residues.…”

Section: Advanced Researchesmentioning

confidence: 99%

Lanthanoid Staining as a Fast Technology of Preparing Microbiological Specimens for Scanning Electron Microscopy

Chebotar¹,

Ia²,

Subbot³

et al. 2017

Sovrem Tehnol Med

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The aim of the investigation was to demonstrate advantages of lanthanoid staining as a novel super-fast technology of preparing microbiological specimens for scanning electron microscopy, and to ground the potentials of using this technology in practical microbiology.Materials and Methods. Microbial cells (bacteria Acinetobacter baumannii, Corynebacterium diphtheriae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumoniae, and fungi Candida albicans), and Staphylococcus aureusbased bacterial biofilms were stained with the preparation based on neodymium chloride and visualized using scanning electron microscopy. Observations were carried out in the mode of low vacuum, images were captured in the mode of detecting back-scattered electrons using a short working section of 4.5-3.0 mm. Specimens were compared to the untreated microorganisms or microbial samples obtained by means of traditional specimen preparation with dehydration and sputtering a gold-platinum coating.Results. The lanthanoid staining technology enabled visualization and evaluation of diagnostically relevant morphological structures of the following microbial cells: A. baumannii, C. diphtheriae, K. pneumoniae, P. aeruginosa, S. aureus, S. pneumoniae, C. albicans, including their relative position, shape, dimensions, presence of capsules and flagella, intracellular structures (polyphosphate inclusions, septum) as well as an extracellular bacterial matrix. The obtained images were characterized by a high contrast, which may be explained by the fact that lanthanoids are capable of providing a high density of electron back-scattering under the impact of the microscope electron beam, resulting in a great brightness of the biological object loci saturated with these elements.Conclusion. Lanthanoid staining of microorganisms is a perspective method of specimen preparation for scanning electron microscopy, which opens the possibility of practical application of this method in diagnostic technologies of clinical microbiology.

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Section: Advanced Researchesmentioning

confidence: 99%

Lanthanoid Staining as a Fast Technology of Preparing Microbiological Specimens for Scanning Electron Microscopy

Chebotar¹,

Ia²,

Subbot³

et al. 2017

Sovrem Tehnol Med

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“…В результате контрастирования на СЭМ маркировались зоны, где кальций изоморфно заменяется на неодим, а также участки локализации энергозависимых процессов с высвобождением фосфат-аниона и последующим формированием нерастворимых фосфатов неодима [13,14]. Подготовленные эпителиальные пласты расправляли поверх специализированной адгезивной углеродной ленты для СЭМ.…”

Section: материал и методыunclassified

“…При этом яркие участки изображения соответствуют насыщенным тяжелыми химическими элементами структурам ткани. Оригинальный метод суправитального лантаноидного контрастирования служит для избирательного насыщения «тяжелыми» лантаноидами отдельных структур и маркирования ими некоторых цепочек клеточного обмена, что делает возможной их последующую визуализацию посредством СЭМ [13,14].…”

Section: результаты и обсуждениеunclassified

SEM visualization of corneal epithelium through lanthanoid staining based on Ca/Nd isomorphous substitution in Ca-dependent molecular systems

Sé¹,

Труфанов

et al. 2016

Vestn. oftal'mol.

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Lanthanoid staining of biopsy material and subsequent SEM enabled detailed visualization of structural features of the corneal epithelium in various pathologies. Due to the Ca/Nd isomorphism we were able to evaluate structural position of the majority of protein molecules engaged in Ca-dependant processes and, consequently, in cytoadherence. Basing on the neodymium distribution within the basal membrane, we have described local effects of different substances on the lamina densa in the projection of basal layer cell borders that occur after unidirectional ultrafiltration. The results confirm the failure of the junctional adhesion complex in recurrent corneal erosion.

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Express-diagnosis of the diseases of the eye and its appendages by scanning electron microscopy with supravital staining

Ia¹,

Kravchik²,

Pak³

et al. 2023

Vestn. oftal'mol.

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Fast and easy method of lanthanoid staining for visualization of cellular ultrastructure and spatial arrangement

Cited by 3 publications

References 4 publications

Lanthanoid Staining as a Fast Technology of Preparing Microbiological Specimens for Scanning Electron Microscopy

Lanthanoid Staining as a Fast Technology of Preparing Microbiological Specimens for Scanning Electron Microscopy

SEM visualization of corneal epithelium through lanthanoid staining based on Ca/Nd isomorphous substitution in Ca-dependent molecular systems

Express-diagnosis of the diseases of the eye and its appendages by scanning electron microscopy with supravital staining

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