A rapid workflow for the characterization of small numbers of unicellular eukaryotes by using correlative light and electron microscopy
Identifikátory výsledku
Kód výsledku v IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11110%2F20%3A10411634" target="_blank" >RIV/00216208:11110/20:10411634 - isvavai.cz</a>
Výsledek na webu
<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=X-SOr_P3Zo" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=X-SOr_P3Zo</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.mimet.2020.105888" target="_blank" >10.1016/j.mimet.2020.105888</a>
Alternativní jazyky
Jazyk výsledku
angličtina
Název v původním jazyce
A rapid workflow for the characterization of small numbers of unicellular eukaryotes by using correlative light and electron microscopy
Popis výsledku v původním jazyce
The limited availability of biological samples hinders phylogenetic efforts to define structural differences among various biological groups. A novel workflow enabling the analysis of protists in low cell numbers by electron microscopy (EM) is described with cysts of Giardia intestinalis, a single-celled eukaryotic parasite. Correlative light and electron microscopy (CLEM) allows for the selection of individual cells and is economical in terms of time and cost. We describe a cyst purification protocol in combination with an adhesive coating for fixation and ultrathin embedding that results in excellent preservation of cell morphology. The application of advanced structural and analytical EM methods, such as high-resolution field emission scanning electron microscopy (FESEM), focused ion beam tomography (FIB/SEM), and energy-dispersive X-ray spectroscopy (EDX) analysis, is demonstrated. The workflow represents a new approach for studying the cellular and organelle architecture of rare and "difficult to culture" microorganisms.
Název v anglickém jazyce
A rapid workflow for the characterization of small numbers of unicellular eukaryotes by using correlative light and electron microscopy
Popis výsledku anglicky
The limited availability of biological samples hinders phylogenetic efforts to define structural differences among various biological groups. A novel workflow enabling the analysis of protists in low cell numbers by electron microscopy (EM) is described with cysts of Giardia intestinalis, a single-celled eukaryotic parasite. Correlative light and electron microscopy (CLEM) allows for the selection of individual cells and is economical in terms of time and cost. We describe a cyst purification protocol in combination with an adhesive coating for fixation and ultrathin embedding that results in excellent preservation of cell morphology. The application of advanced structural and analytical EM methods, such as high-resolution field emission scanning electron microscopy (FESEM), focused ion beam tomography (FIB/SEM), and energy-dispersive X-ray spectroscopy (EDX) analysis, is demonstrated. The workflow represents a new approach for studying the cellular and organelle architecture of rare and "difficult to culture" microorganisms.
Klasifikace
Druh
J<sub>imp</sub> - Článek v periodiku v databázi Web of Science
CEP obor
—
OECD FORD obor
10606 - Microbiology
Návaznosti výsledku
Projekt
<a href="/cs/project/LTAB19004" target="_blank" >LTAB19004: Mitozomy Giardia intestinalis: unikátní redukované mitochondrie v kontexu životního cyklu mikroaerofilního parazitního prvoka</a><br>
Návaznosti
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Ostatní
Rok uplatnění
2020
Kód důvěrnosti údajů
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Údaje specifické pro druh výsledku
Název periodika
Journal of Microbiological Methods
ISSN
0167-7012
e-ISSN
—
Svazek periodika
172
Číslo periodika v rámci svazku
May
Stát vydavatele periodika
NL - Nizozemsko
Počet stran výsledku
6
Strana od-do
105888
Kód UT WoS článku
000528192100007
EID výsledku v databázi Scopus
2-s2.0-85082645828