Enhanced electron microscopy imaging for a detailed structural study of alginate hydrogel containing the encapsulated cells

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081731%3A_____%2F25%3A00638974" target="_blank" >RIV/68081731:_____/25:00638974 - isvavai.cz</a>

Výsledek na webu

<a href="https://www.sciencedirect.com/science/article/pii/S0144861725010240" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0144861725010240</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.carbpol.2025.124239" target="_blank" >10.1016/j.carbpol.2025.124239</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Enhanced electron microscopy imaging for a detailed structural study of alginate hydrogel containing the encapsulated cells

Popis výsledku v původním jazyce

Hydrogels are widely applicable in medicine, biotechnology, etc. A specific example is bacterial alginate produced by plant growth-promoting rhizobacterium Azotobacter vinelandii, which encapsulates the cells within its hydrogel network, offering promising applications in agriculture. To better understand the properties and behaviour of hydrogel, it is important to study its architecture. However, due to high-water content and fine structure of hydrogel samples, their preparation for electron microscopy is challenging. In this study, we developed an optimised protocol for preparing complex samples of A. vinelandii cells encapsulated in alginate hydrogel for imaging by low-voltage scanning transmission electron microscopy. Our approach addresses structural instability and artefact formation typically encountered during sample dehydration and staining. We demonstrated that careful timing of CaCl2 addition, after initial fixation, is essential. Equally important is the careful selection of its concentration to maintain hydrogel integrity while preserving cellular morphology. The inclusion of lead citrate staining step enhanced the contrast within hydrogel matrix, allowing for improved visualisation of fine details. This refined protocol enables high-resolution imaging of alginate-based hydrogels containing embedded cells while preserving the key structural features. It is compatible with various transmission electron microscopy techniques and may be adapted for use with other soft biomaterials.

Název v anglickém jazyce

Enhanced electron microscopy imaging for a detailed structural study of alginate hydrogel containing the encapsulated cells

Popis výsledku anglicky

Hydrogels are widely applicable in medicine, biotechnology, etc. A specific example is bacterial alginate produced by plant growth-promoting rhizobacterium Azotobacter vinelandii, which encapsulates the cells within its hydrogel network, offering promising applications in agriculture. To better understand the properties and behaviour of hydrogel, it is important to study its architecture. However, due to high-water content and fine structure of hydrogel samples, their preparation for electron microscopy is challenging. In this study, we developed an optimised protocol for preparing complex samples of A. vinelandii cells encapsulated in alginate hydrogel for imaging by low-voltage scanning transmission electron microscopy. Our approach addresses structural instability and artefact formation typically encountered during sample dehydration and staining. We demonstrated that careful timing of CaCl2 addition, after initial fixation, is essential. Equally important is the careful selection of its concentration to maintain hydrogel integrity while preserving cellular morphology. The inclusion of lead citrate staining step enhanced the contrast within hydrogel matrix, allowing for improved visualisation of fine details. This refined protocol enables high-resolution imaging of alginate-based hydrogels containing embedded cells while preserving the key structural features. It is compatible with various transmission electron microscopy techniques and may be adapted for use with other soft biomaterials.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

20903 - Bioproducts (products that are manufactured using biological material as feedstock) biomaterials, bioplastics, biofuels, bioderived bulk and fine chemicals, bio-derived novel materials

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2025

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Carbohydrate Polymers

ISSN

0144-8617

e-ISSN

1879-1344

Svazek periodika

368

Číslo periodika v rámci svazku

2

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

9

Strana od-do

124239

Kód UT WoS článku

001561887400001

EID výsledku v databázi Scopus

2-s2.0-105013584964