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Transmission electron microscopic (TEM) overview of a glomerular section. Shown section is a consecutive section to the RCM section (MP9526). Specific RCM reflection is correlated with staining of albumin in this TEM overview. Chronic infusion of hyaluronidase leads to degradation of the glycocalyx. This results in the presence of albumin (dark black spots) bound to podocyte foot processes and on podocyte body and parietal epithelial cells. The sample has been imaged at 1.2 nm pixel-1 resolution over an area of 124 x 170 µm2.
Full resolution image description
Tiff file of a transmission electron microscopic (TEM) overview of a glomerular section from the HA treatment group. Shown section is a consecutive section to the RCM section (MP9526). Specific RCM reflection is correlated with staining of albumin in this TEM overview. The sample has been imaged at 1.2 nm pixel-1 resolution over an area of 124 x 170 µm2.
File format
TIFF
Volume_dimension
105984, 145408, 1
Volume scale
0.0012, 0.0012, 0.1
License
Attribution Only: This image is licensed under a Creative Commons Attribution License. View License Deed | View Legal Code

CCDB:9525*  Cite 
Project: P2133
Project name
Role of the endothelial glycocalyx in renal glomerular and tubular function
Description
Determine the role of the endothelial glycocalyx in the glomerular filtration barrier.
Funding agency
Dutch Kidney Foundation
Leader(s)
Dr. Bernard M. van den Berg
Prof Dr Ton J. Rabelink
Collaborator(s)
Martijn J.C. Dane
Bernard M. van den Berg
M. Cristina Avramut
Frank G.A. Faas
Johan van der Vlag
Angelique L.W.M.M. Rops
Raimond B.G. Ravelli
A.J. Koster
Anton Jan van Zonneveld
Hans Vink
Ton J. Rabelink
Start date
09-01-2009
End date
09-01-2009
 
Experiment
Experiment ID
9515
Experiment date
01-01-2011
Title
Perturbation of the glomerular endothelial surface layer results in albumin filtration
Purpose
Determine the effect of hyaluronidase on the filtration of albumin over the glomerular filtration barrier
Experimenter(s)
Martijn Dane
Bernard van den Berg
Cristina Avramut
Angelique Rops
Microscopy product
Microscopy product ID
9525
Instrument
Transmission Electron Microscope
Microscopy type
TEM
Product type
MOSAIC
Image basename
D2_RK_Ab1to100_18500x_stitch
Spatial Axis Image Size Pixel Size
X 124px 1.2 nm/pixels
Y 170px 1.2 nm/pixels
Subject
Species
Mouse
Scientific name
mus musculus
Strain
C57Bl/6
Group by
Treatment
Treatment
Mice were treated with active hyaluronidase via an osmotic minipump for either 2 or 4 weeks.
Age
3 months
Age class
Adult
Tissue section
Anatomical location
Kidney
Microtome
Reichert Ultracut S (Leica)
Thickness
0.1 µm
Specimen description
Organ
kidney
Imaging parameters
Type
Electron microscopy product
Magification
18500
Accelerating voltage
120 KV
Specimen preparation
Protocol used
Kidney tissue was dissected at 100 um thickness with a vibratome. Part of the slices are blocked with 10 percent normal goat serum and 0.3 percent Triton 100 in PBS for 30 minutes on ice followed by incubation o/n at 4 degrees C with horseradish peroxidase-conjugated goat anti-mouse albumin (Bethyl Labs, USA, diluted to 1/400) in 1 percent heat inactivated normal goat serum in PBS. After washing, about 1mL of 3,3'-diaminobenzidine reaction solution (15 mL DAB solution + 10-15 uL H2O2, 30 percent, (Dako, Denmark)) was added, incubated for 30 minutes at 4 degrees C, washed again and incubated for 1 hour with 1.5 percent GA and 1% PFA in cacodylate, rinsed in cacodylate and postfixated in 1 percent osmium tetroxide + 1.5 percent potassium ferrocyanide. Samples were dehydrated and processed further into epon. Sequential 100 nm sections were mounted on a copper slot grid covered with Formvar support film and a 3 nm carbon coating for TEM, and on a water drop on a clean glass slide for RCM.20 The sample was mounted with immersion oil (Immersol 518F, Zeiss) on a RCM-adapted microscope (reflection contrast device RV; Leica, Wetzlar Germany). Images were recorded with a 100X, 1.25 NA objective. TEM sections were stained with an aqueous solution of lead citrate and uranyl acetate before visualization and data was collected at an acceleration voltage of 120 kV,on a Tecnai 12 (BioTWIN) transmission electron microscope, equipped with an Eagle CCD camera (FEI company, Eindhoven, the Netherlands).
Imaging product type
Type
Mosaic
X position
69 tiles
Y position
95 tiles