Microbiology & Virology
The world of microbiology can be as wonderful as it is colourful. Huygens helps scientists improve their microscopic imaging and increase their scientific impact. Explore the benefits of deconvolution on our cell biology page or push your resolution to the next level on our super-res page. Check out below protocol for quantitative imaging of viral particles.
Protocol: Quantitative analysis of viral particles
Image description:
Lyngba cyanobacteria. Taken on an SP5 Leica confocal laser scanning microscope and restored with Huygens. Specimens are stained with acridine orange, but much of the color is due to autofluorescence. Image courtesy of Tagide deCarvalho PhD, University of Maryland Baltimore County, United States
Lyngba cyanobacteria. Taken on an SP5 Leica confocal laser scanning microscope and restored with Huygens. Specimens are stained with acridine orange, but much of the color is due to autofluorescence. Image courtesy of Tagide deCarvalho PhD, University of Maryland Baltimore County, United States
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Protocol: Quantitative analysis of viral particles
- Confocal z-stack was recorded on a Zeiss LSM700 microscope with 63X 1.4NA planApochromat oil objective.
- Image was loaded into Huygens, for confocal images all image parameters are automatically read in. No additional user input is required.
- A measured Point-Spread-Function (PSF) was obtained using Huygens PSF distiller from recordings of immobilised fluorescent beads. Huygens comes with a powerful theoretical PSF calculation engine, which by default generates reliable and accurate deconvolution results. For that bit of extra, a measured PSF additionally accounts for defects specific to your system, improving resolution and intensity-information in your images.
- Image is deconvolved using Huygens Professional.
- Image is segmented using Huygens Object analyzer and visualised using Huygens surface renderer. Segmentation occurred using a watershed algorithm, allowing for intelligent object separation. Objects were coloured randomly using rainbow colouring.
- Statistics on viral particles brightnesses and sizes are obtained from the object segmentation. Data can be exported to common text formats .csv (excel), text or Matlab files for further analysis.
Release of GFP-containing viral particles from the infected cell. Image courtesy of Dr. Elena Frolova, Department of Microbiology, University of Alabama at Birmingham, United States.
Microbiology & Virology Image Gallery
Image of cyanobacteria mat/biofilm. Taken on an SP5 Leica confocal laser scanning microscope and restored with Huygens. Specimens are stained with acridine orange, but much of the color is due to autofluorescence. Image kindly provided by Tagide deCarvalho PhD, University of Maryland Baltimore County, United States.
Image shows a growing community of bacteria Vibrio cholerae, the pathogen for cholerae. Bacteria adopt a social life form known as biofilm, where they stay together and secrete chemicals to build a "home" for survival. This Nikon spinning disc confocal image was deconvolved with a theoretical PSF in Huygens and finally color coded for depth. Image kindly provided by Dr. Jing Yan, Princeton University, United States.