Comparison of TEM sample preparation methods for immunogold labeling

Christian GRECHHAMER

Bakalářská práce

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Abstract:

The tracking of antigens in biological samples can be achieved by labeling them with gold-gonjugated antibodies (immunogold labeling). This thesis gives an overview of several sample preparation methods for TEM and investigates their effects on the binding efficiency of antibodies to certain antigens. For this reason, protein samples were processed according to four different protocols, cut into ultra-thin sections and labeled with primary and secondary antibodies. The micrographs, taken during observation with a TEM, were analysed quantitatively and the obtained data was statistically evaluated.

Abstract:

The tracking of antigens in biological samples can be achieved by labeling them with gold-gonjugated antibodies (immunogold labeling). This thesis gives an overview of several sample preparation methods for TEM and investigates their effects on the binding efficiency of antibodies to certain antigens. For this reason, protein samples were processed according to four different protocols, cut into ultra-thin sections and labeled with primary and secondary antibodies. The micrographs, taken during observation with a TEM, were analysed quantitatively and the obtained data was statistically evaluated.

Keywords

Transmission Electron Microscopy, immunogold labeling, microwave-assisted sample preparation, high pressure freezing, super quick freeze substitution, microtomy, gold quantitation, labeling density

Jazyk práce: angličtina

Datum vytvoření / odevzdání či podání práce: 15. 5. 2017

Obhajoba závěrečné práce

• Vedoucí: RNDr. Marie Vancová, Ph.D.

JIHOČESKÁ UNIVERZITA V ČESKÝCH BUDĚJOVICÍCH

Přírodovědecká fakulta

Bakalářský studijní program / obor:
Biochemistry / Biological Chemistry