alexa Utilization of Non-Specific Staining to Visualise Liver Vasculature in Mice | Open Access Journals
ISSN: 2157-7099
Journal of Cytology & Histology
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Utilization of Non-Specific Staining to Visualise Liver Vasculature in Mice

Eva Szunyogova1,2 and Simon H Parson1*

1University of Aberdeen, Institute of Medical Sciences, Aberdeen, United Kingdom

2Euan MacDonald Centre for Motor Neurone Disease Research, Edinburgh, United Kingdom

*Corresponding Author:
Simon H Parson
University of Aberdeen
Institute of Medical Sciences
Aberdeen, United Kingdom
Tel: 44 (0)1224 274328
E-mail: [email protected]

Received Date: August 02, 2015 Accepted Date: August 04, 2015 Published Date: August 06, 2015

Citation: Szunyogova E (2015) Utilization of Non-Specific Staining to Visualise Liver Vasculature in Mice. J Cytol Histol 2015, 6:i111. doi.org/10.4172/2157-7099.1000i111

Copyright: © 2015 Szunyogova E, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Description

Histological identification of morphological alterations are key indicators of functional defects. For example, liver disease which compromises perfusion and consequently function, is very often associated with abnormal tissue microvasculature. Traditionally an anti-platelet endothelial cell adhesion molecule-1 (PECAM-1/ CD31) antibody is utilized to visualize the vasculature, however in mouse liver this does not show the entire vasculature. The sinusoidal endothelial cells are a unique subtype of endothelial cells more closely structurally and functionally related to lymphatic vessels. Unlike other vessels, during postnatal development liver sinusoidal endothelial cells (LSECs) downregulate PECAM-1 levels to facilitate their high permeability [1]. Therefore to evaluate sinusoidal defects, another marker is required. Here we present a simple, effective and cheap method to visualise the entire liver vasculature utilization ‘nonspecific’ staining by an Fc antibody fragment (Figure 1).

cytology-histology-fc-fragment-staining

Figure 1: Fc fragment staining of mouse liver at postnatal day 20. A) Representative micrographs of Fc fragment staining of vasculature (green: left panel) and B) combined with DAPI stain for nuclei (blue: right panel). Scale bar=50 μm.

Liver was harvested from mice sacrificed at postnatal day 20 (P20) and fixed for 4 hours in 4% paraformaldehyde 0.1 M phosphate buffered saline (PBS). After fixation, the liver was washed in PBS and placed into 30% sucrose solution (made up in PBS) for subsequent OCT embedding. Liver sections (5 μm) were air dried, washed in PBS and left in blocking solution (0.2% bovine serum albumin (BSA), 0.5%Triton X-100 (TX100) in PBS) for 2 hours. Once blocked, the slides were immunostained only with secondary antibody, Alexa 488- goat anti-mouse IgG (Abcam, ab150113). All sections were visualised using a Nikon eclipse e400 microscope (20X objective) and its images captured using QICAM Fast 1394 camera and Improvision Velocity 4 image capture software.

The Fc fragment is most likely part of the IgG receptor expressed on LSECs that eliminates small immune complexes, thereby controlling complex-mediated autoimmune disease [2]. This simple process then visualises the entire liver vasculature in a single step.

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