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Development and application of advanced electron microscopy techniques for materials in biological systems

Nevsten, Pernilla LU (2008)
Abstract
Theoretically, electron microscopy and scanning probe microscopy offer information on a scale beyond the optical microscope. However, the quality of the micrograph and the obtained information is limited by the sample preparation. Each sample has its own characteristics that need special precautions, meaning that sample preparation is a continuously developing science. In this thesis several microscopy techniques are further developed and refined, and the problems involved are discussed both in general terms and more specifically for:



Immunogold labelling of budded baculovirus, was performed to support the expression of a test protein, in a new display system for screening of proteins.

Immunogold labelling was... (More)
Theoretically, electron microscopy and scanning probe microscopy offer information on a scale beyond the optical microscope. However, the quality of the micrograph and the obtained information is limited by the sample preparation. Each sample has its own characteristics that need special precautions, meaning that sample preparation is a continuously developing science. In this thesis several microscopy techniques are further developed and refined, and the problems involved are discussed both in general terms and more specifically for:



Immunogold labelling of budded baculovirus, was performed to support the expression of a test protein, in a new display system for screening of proteins.

Immunogold labelling was performed to locate hormone-sensitive lipase (HSL) within the beta cells in the islets of Langerhans. HSL was shown be located to the insulin storing granules.

TEM was used to investigate the nature of the first precipitate formed in a solution designed to mimic the inorganic composition of the blood plasma of dialysis patients, i.e. with increased phosphate concentration.

TEM and SEM were used to analyze the effects of modifying α-TCP, to be used as bone support material, by increasing milling time of the α-TCP particles or by substitution with silicon or strontium.

SEM was used to put further light on release mechanisms for polymer coated pellets to be used in multiple-unit pharmaceutical devices.

The possibility of correlative imaging by AFM, TEM and SEM of the exact same sample object was investigated on stained and unstained baculovirus.

For further development of general microscopy protocols, different grid designs were investigated by SEM and commonly used treatment protocols, for modifying the hydrophilic properties of supports, were ranked by measuring the water contact angle on carbon after treatment.

For cryo-TEM, the effect of adding a surface active stain to a virus-suspension on the frequency of virus particles in the vitrified film was evaluated by surface tension measurements. (Less)
Please use this url to cite or link to this publication:
author
supervisor
opponent
  • Pr. Hebert, Hans, Dept. of Bioscience and Nutrition, Karolinska Institutet, Stockholm, Sweden
organization
publishing date
type
Thesis
publication status
published
subject
keywords
multiple-unit drug, EFTEM, EELS, cyanine dye, SSAT, cryo-TEM, virus, TEM, apoptosis, SEM, AFm, α-TCP, apatite, XEDS, TEM grid, microscopy supports
defense location
Center of Chemistry and Chemical Engineering, lecture hall K:C
defense date
2008-04-18 13:15
ISBN
978-97-7422-191-6
language
English
LU publication?
yes
id
89d84073-9204-4f7d-8aa3-b1a907c8d2bb (old id 1049472)
date added to LUP
2008-03-25 08:50:37
date last changed
2016-09-19 08:45:16
@misc{89d84073-9204-4f7d-8aa3-b1a907c8d2bb,
  abstract     = {Theoretically, electron microscopy and scanning probe microscopy offer information on a scale beyond the optical microscope. However, the quality of the micrograph and the obtained information is limited by the sample preparation. Each sample has its own characteristics that need special precautions, meaning that sample preparation is a continuously developing science. In this thesis several microscopy techniques are further developed and refined, and the problems involved are discussed both in general terms and more specifically for:<br/><br>
<br/><br>
Immunogold labelling of budded baculovirus, was performed to support the expression of a test protein, in a new display system for screening of proteins.<br/><br>
Immunogold labelling was performed to locate hormone-sensitive lipase (HSL) within the beta cells in the islets of Langerhans. HSL was shown be located to the insulin storing granules. <br/><br>
TEM was used to investigate the nature of the first precipitate formed in a solution designed to mimic the inorganic composition of the blood plasma of dialysis patients, i.e. with increased phosphate concentration. <br/><br>
TEM and SEM were used to analyze the effects of modifying α-TCP, to be used as bone support material, by increasing milling time of the α-TCP particles or by substitution with silicon or strontium.<br/><br>
SEM was used to put further light on release mechanisms for polymer coated pellets to be used in multiple-unit pharmaceutical devices.<br/><br>
The possibility of correlative imaging by AFM, TEM and SEM of the exact same sample object was investigated on stained and unstained baculovirus.<br/><br>
For further development of general microscopy protocols, different grid designs were investigated by SEM and commonly used treatment protocols, for modifying the hydrophilic properties of supports, were ranked by measuring the water contact angle on carbon after treatment.<br/><br>
For cryo-TEM, the effect of adding a surface active stain to a virus-suspension on the frequency of virus particles in the vitrified film was evaluated by surface tension measurements.},
  author       = {Nevsten, Pernilla},
  isbn         = {978-97-7422-191-6},
  keyword      = {multiple-unit drug,EFTEM,EELS,cyanine dye,SSAT,cryo-TEM,virus,TEM,apoptosis,SEM,AFm,α-TCP,apatite,XEDS,TEM grid,microscopy supports},
  language     = {eng},
  title        = {Development and application of advanced electron microscopy techniques for materials in biological systems},
  year         = {2008},
}