Dr. David M. Haaland Profile

Dr. David M. Haaland

Senior Scientist at Spectral Resolutions

SPIE Involvement:

Author

Proceedings Article | 15 October 2007 Paper

Hyperspectral confocal fluorescence imaging of cells

David Haaland, Howland Jones, Michael Sinclair, Bryan Carson, Catherine Branda, Jens Poschet, Roberto Rebeil, Bing Tian, Ping Liu, Allan Brasier

Proceedings Volume 6765, 676509 (2007) https://doi.org/10.1117/12.738152

KEYWORDS: Microscopes, Luminescence, Green fluorescent protein, Hyperspectral imaging, Confocal microscopy, Composites, Imaging systems, Proteins, Image analysis, Live cell imaging

Read Abstract +

Confocal fluorescence imaging of biological systems is an important method by which researchers can investigate molecular processes occurring in live cells. We have developed a new 3D hyperspectral confocal fluorescence microscope that can further enhance the usefulness of fluorescence microscopy in studying biological systems. The new microscope can increase the information content obtained from the image since, at each voxel, the microscope records 512 wavelengths from the emission spectrum (490 to 800 nm) while providing optical sectioning of samples with diffraction-limited spatial resolution. When coupled with multivariate curve resolution (MCR) analyses, the microscope can resolve multiple spatially and spectrally overlapped emission components, thereby greatly increasing the number of fluorescent labels, relative to most commercial microscopes, that can be monitored simultaneously. The MCR algorithm allows the "discovery" of all emitting sources and estimation of their relative concentrations without cross talk, including those emission sources that might not have been expected in the imaged cells. In this work, we have used the new microscope to obtain time-resolved hyperspectral images of cellular processes. We have quantitatively monitored the translocation of the GFP-labeled RelA protein (without interference from autofluorescence) into and out of the nucleus of live HeLa cells in response to continuous stimulation by the cytokine, TNFα. These studies have been extended to imaging live mouse macrophage cells with YFP-labeled RelA and GFP-labeled IRF3 protein. Hyperspectral imaging coupled with MCR analysis makes possible, for the first time, quantitative analysis of GFP, YFP, and autofluorescence without concern for cross-talk between emission sources. The significant power and quantitative capabilities of the new hyperspectral imaging system are further demonstrated with the imaging of a simple fluorescence dye (SYTO 13) traditionally used to stain the nucleus of live cells. We will demonstrate the microscope system's ability to actually discover and quantify the presence of two separate SYTO 13 fluorescent species shifted in wavelength by only a few nm. These two emission components exhibit very different spatial distributions in macrophage cells (i.e., nucleus vs. cytoplasm + nucleus). Two highly overlapped autofluorescence components in addition to the two SYTO 13 components were also observed, and the spatial distributions of the two autofluorescence components were quantitatively mapped throughout the cells in three dimensions.

Proceedings Article | 27 February 2007 Paper

Exploring membrane protein dynamics by multicolor single quantum dot imaging using wide field, TIRF, and hyperspectral microscopy

Diane Lidke, Nicholas Andrews, Janet Pfeiffer, Howland D.T. Jones, Michael Sinclair, David Haaland, Alan Burns, Bridget Wilson, Janet Oliver, Keith Lidke

Proceedings Volume 6448, 64480Y (2007) https://doi.org/10.1117/12.718464

KEYWORDS: Proteins, Microscopes, Microscopy, Hyperspectral imaging, Diffusion, Receptors, Imaging systems, Quantum dots, Luminescence, Particles

Read Abstract +

Proceedings Article | 21 February 2006 Paper

Imaging multiple endogenous and exogenous fluorescent species in cells and tissues

Jerilyn Timlin, Linda Nieman, Howland Jones, Michael Sinclair, David Haaland, John Guzowski

Proceedings Volume 6088, 608805 (2006) https://doi.org/10.1117/12.647228

KEYWORDS: Tissues, Hyperspectral imaging, Microscopes, Glasses, Luminescence, Image analysis, Brain, Biological research, Neuroimaging, Optical filters

Read Abstract +

Proceedings Article | 15 October 2004 Paper

Multivariate curve resolution for the analysis of remotely-sensed thermal infrared hyperspectral images

Chris Stork, Michael Keenan, David Haaland

Proceedings Volume 5546, (2004) https://doi.org/10.1117/12.559604

KEYWORDS: Absorption, Hyperspectral imaging, Transmittance, Data modeling, Atmospheric modeling, Infrared imaging, Thermography, Infrared radiation, Image resolution, Atmospheric sensing

Read Abstract +

Proceedings Article | 2 July 2003 Paper

Multivariate curve resolution for hyperspectral image analysis: applications to microarray technology

David Haaland, Jerilyn Timlin, Michael Sinclair, Mark Van Benthem, M. Juanita Martinez, Anthony Aragon, Margaret Werner-Washburne

Proceedings Volume 4959, (2003) https://doi.org/10.1117/12.477945

KEYWORDS: Scanners, Glasses, Axons, Hyperspectral imaging, Luminescence, Image analysis, Fluorescent markers, Image resolution, Optical filters, Laser scanners

Read Abstract +

Proceedings Article | 8 November 2002 Paper

Algorithms for constrained linear unmixing with application to the hyperspectral analysis of fluorophore mixtures

Michael Keenan, Jerilyn Timlin, Mark Van Benthem, David Haaland

Proceedings Volume 4816, (2002) https://doi.org/10.1117/12.451662

KEYWORDS: Luminescence, Hyperspectral imaging, Principal component analysis, Statistical analysis, Laser scattering, Sensors, Remote sensing, Fluorescent markers, Image sensors, Signal detection

Read Abstract +

Proceedings Article | 22 February 2002 Paper

Infrared ATR: a probe for cellular activation

Jerilyn Timlin, Laura Martin, M. Kathleen Alam, David Haaland, Kristen Garrison, C. Richard Lyons, Brian Hjelle

Proceedings Volume 4577, (2002) https://doi.org/10.1117/12.455749

KEYWORDS: Crystals, Infrared spectroscopy, Attenuated total reflectance, Proteins, Infrared radiation, Absorbance, Spectroscopy, FT-IR spectroscopy, Sensors, Water

Read Abstract +

Proceedings Article | 31 January 1994 Paper

Multivariate calibration applied to near-infrared spectroscopy for the quantitative analysis of dilute aqueous solutions

David Haaland, Howland Jones

Proceedings Volume 2089, (1994) https://doi.org/10.1117/12.166659

KEYWORDS: Calibration, Statistical analysis, Near infrared spectroscopy, Urea, Quantitative analysis, Statistical modeling, Tissue optics, FT-IR spectroscopy, In vitro testing, Spectral calibration

Read Abstract +

Proceedings Article | 1 March 1992 Paper

Multivariate calibration applied to the quantitative analysis of infrared spectra

David Haaland

Proceedings Volume 1575, (1992) https://doi.org/10.1117/12.56281

KEYWORDS: Calibration, Infrared radiation, Spectral calibration, Factor analysis, Glucose, Blood, Statistical analysis, Infrared spectroscopy, Spectroscopy, Phosphorus

Read Abstract +

Proceedings Article | 1 March 1991 Paper

Fourier transform infrared spectrophotometry for thin film monitors: computer and equipment integration for enhanced capabilities

J. Neal Cox, J. Sedayao, Gurmeet Shergill, R. Villasol, David Haaland

Proceedings Volume 1392, (1991) https://doi.org/10.1117/12.48957

KEYWORDS: FT-IR spectroscopy, Data modeling, Spectrophotometry, Thin films, Infrared radiation, Phosphorus, Calibration, Boron, Statistical analysis, Silicon films

Read Abstract +

Showing 5 of 10 publications

Conference Committee Involvement (4)

Next-Generation Spectroscopic Technologies V

23 April 2012 | Baltimore, Maryland, United States

Next-Generation Spectroscopic Technologies IV

25 April 2011 | Orlando, Florida, United States

Next-Generation Spectroscopic Technologies III

5 April 2010 | Orlando, Florida, United States

Next-Generation Spectroscopic Technologies

10 September 2007 | Boston, MA, United States

Keywords/Phrases

Search In:

Publication Years