The granules
were stained and mounted in coverwell chambers with a 1 mm spacer
in order to avoid squeezing of the samples. Glycoconjugates of the
anammox granules were examined by means of fluorescence lectin bar-coding.33 (link) Thus, all commercially available lectins (FITC
or Alexa488) were applied as an individual probe to one granule. For
3d imaging a TCS SP5X confocal laser scanning microscope (Leica, Germany)
was employed. The upright microscope was equipped with a super continuum
light source and controlled by the software LAS AF 2.4.1. The confocal
data sets were recorded by using 25× NA 0.95 and 63× NA
1.2 water immersion lenses. Excitation was at 490 nm (laser power
70% at laser, 50% in software), and emission signals were detected
simultaneously with two photomultipliers from 485 to 495 nm (reflection)
and 505–600 nm (fluorescence). Image data sets were deconvolved
with Huygens version 16.05 using the CMLE algorithm (SVI, The Netherlands)
and projected with Imaris version 9.1.2 (Bitplane, Switzerland).
were stained and mounted in coverwell chambers with a 1 mm spacer
in order to avoid squeezing of the samples. Glycoconjugates of the
anammox granules were examined by means of fluorescence lectin bar-coding.33 (link) Thus, all commercially available lectins (FITC
or Alexa488) were applied as an individual probe to one granule. For
3d imaging a TCS SP5X confocal laser scanning microscope (Leica, Germany)
was employed. The upright microscope was equipped with a super continuum
light source and controlled by the software LAS AF 2.4.1. The confocal
data sets were recorded by using 25× NA 0.95 and 63× NA
1.2 water immersion lenses. Excitation was at 490 nm (laser power
70% at laser, 50% in software), and emission signals were detected
simultaneously with two photomultipliers from 485 to 495 nm (reflection)
and 505–600 nm (fluorescence). Image data sets were deconvolved
with Huygens version 16.05 using the CMLE algorithm (SVI, The Netherlands)
and projected with Imaris version 9.1.2 (Bitplane, Switzerland).
