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Photophysical and Chemical Approaches to Cellular Biophysics
KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
2008 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

The central theme in this thesis is reversibility. Two main attempts has been made to approach reversibility in cellular systems from both chemical and physical points of view. Reversibility of immunolabeling of proteins on the cell surface has been adressed by development of new fluorescent substances optimized for CALI (Chromophore-Assisted Laser Inactivation of protein). Aluminum phthalocyanine (AlPc) is here identified to be a good candidate for a new generation of fluorophores for efficient hydroxyl radical generation. It is shown that cells can be reversibly labeled with antibody-AlPc conjugates. In experiments on living cells the AlPcs were not only active as classic fluorophores but also as photocatalytic substances with destaining properties. Reversibility of cell immobilization is also reported, where cells cultured in microstructures were immobilized and 3D supported using hydrogels. Hydrogel formulation and application was optimized to achieve a system where both viability and ease of use was satisfied. Gel reversibility was actualized with pH and enzyme treatment. The developped method offers the possibility of stop flow culturing cells in controlled and reusable 3D environments.

Place, publisher, year, edition, pages
Stockholm: KTH , 2008. , v, 26 p.
Series
Trita-FYS, ISSN 0280-316X ; 2008:54
National Category
Biophysics
Identifiers
URN: urn:nbn:se:kth:diva-10097ISBN: 978-91-7412-200-5 OAI: oai:DiVA.org:kth-10097DiVA: diva2:208187
Presentation
2008-12-18, FA 32, AlbaNova Universitetscentrum, Roslagstullsbacken 21, Stockholm, 11:00 (English)
Opponent
Supervisors
Note
QC 20101102Available from: 2009-03-16 Created: 2009-03-16 Last updated: 2010-11-02Bibliographically approved
List of papers
1. Photophysics and photochemistry of zinc, aluminium and tin octakis (benzylthio) phthalocyanines
Open this publication in new window or tab >>Photophysics and photochemistry of zinc, aluminium and tin octakis (benzylthio) phthalocyanines
2008 (English)Report (Other academic)
Series
Trita-FYS, ISSN 0280-316X ; 2008:44
Keyword
Zinc, aluminum and tin benzylthio phthalocyanines, singlet oxygen, triplet yield, triplet lifetime
National Category
Biophysics
Identifiers
urn:nbn:se:kth:diva-10051 (URN)
Note
QC 20101102Available from: 2009-03-10 Created: 2009-03-10 Last updated: 2010-11-02Bibliographically approved
2. Characterization studies of aluminum phthalocyanine binding to antibodies from SKBR 3 cell line
Open this publication in new window or tab >>Characterization studies of aluminum phthalocyanine binding to antibodies from SKBR 3 cell line
2008 (English)Report (Other academic)
Series
Trita-FYS, ISSN 0280-316X ; 2008:45
Keyword
antibody, photocleavage, confocal microscopy, FRAP, CALI, photobleaching, photochemical reaction
National Category
Biophysics
Identifiers
urn:nbn:se:kth:diva-10043 (URN)
Note
QC 20101102Available from: 2009-03-09 Created: 2009-03-09 Last updated: 2010-11-02Bibliographically approved
3. Self-assembling Fmoc dipeptide hydrogel for in situ 3D cell culturing
Open this publication in new window or tab >>Self-assembling Fmoc dipeptide hydrogel for in situ 3D cell culturing
2007 (English)In: BMC Biotechnology, ISSN 1472-6750, Vol. 7, no 88Article in journal (Refereed) Published
Abstract [en]

Background: Conventional cell culture studies have been performed on 2D surfaces, resulting in flat, extended cell growth. More relevant studies are desired to better mimic 3D in vivo tissue growth. Such realistic environments should be the aim of any cell growth study, requiring new methods for culturing cells in vitro. Cell biology is also tending toward miniaturization for increased efficiency and specificity. This paper discusses the application of a self-assembling peptide-derived hydrogel for use as a 3D cell culture scaffold at the microscale.

Results: Phenylalanine derivative hydrogel formation was seen to occur in multiple dispersion media. Cells were immobilized in situ within microchambers designed for cell analysis. Use of the highly biocompatible hydrogel components and simplistic procedures significantly reduced the cytotoxic effects seen with alternate 3D culture materials and microstructure loading methods. Cells were easily immobilized, sustained and removed from microchambers. Differences in growth morphology were seen in the cultured cells, owing to the 3-dimentional character of the gel structure. Degradation improved the removal of hydrogel from the microstructures, permitting reuse of the analysis platforms.

Conclusion: Self-assembling diphenylalanine derivative hydrogel provided a method to dramatically reduce the typical difficulties of microculture formation. Effective generation of patterned 3D cultures will lead to improved cell study results by better modeling in vivo growth environments and increasing efficiency and specificity of cell studies. Use of simplified growth scaffolds such as peptide-derived hydrogel should be seen as highly advantageous and will likely become more commonplace in cell culture methodology.

Keyword
Cell analysis, Cell culture scaffold, Dipeptide hydrogel
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-10044 (URN)10.1186/1472-6750-7-88 (DOI)000252971600001 ()2-s2.0-39049149107 (Scopus ID)
Note
QC 20100806Available from: 2009-03-09 Created: 2009-03-09 Last updated: 2010-11-19Bibliographically approved

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CiteExportLink to record
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Citation style
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