Journal: Molecular therapy : the journal of the American Society of Gene Therapy

Article Title: Gene transfer corrects acute GM2 gangliosidosis--potential therapeutic contribution of perivascular enzyme flow.

doi: 10.1038/mt.2012.44

Figure Lengend Snippet: Figure 6 Inflammatory response in the presence of residual storage and emerging disease in nontargeted tissues. (a) The intensity and number of cells staining for the microglial marker, CD68, and astrocytic marker, GFAP, correlated with local glycoconjugate abundance (stained magenta with periodic acid-Schiff (PAS) reagent). Displayed are sections from an Sx2-injected Sandhoff (SD) mouse, killed at 696 days and viewed in Supplementary Videos S4 and S5 at 12 and 20 months of age. The ninth cerebellar lobule with few PAS-stained neurons (hash area and inset in left panel) had few cells labeled with CD68 (arrowheads in left panel) and GFAP (arrowhead in left panel). In contrast, the flocculus lobe with many intensely PAS-stained neurons (asterisk area and inset in right panel) had many cells labeled with CD68 (arrowheads, right panel) and GFAP (arrow- heads, right panel). Globular microglia (arrows in left bottom panel) and astrocytes (arrows, right bottom panel) are intensely stained. (b) Abundant periodic acid-Schiff (PAS)-stained glycoconjugate was found in retinal ganglion cell layer (GL) in 4-month untreated SD [SD (UT)] compared with 2-year-old wild type (WT). In S+C-treated 2 year-old SD [SD (T)] staining was found in GL and inner nuclear (INL) layers. (c) SD (T) mouse endothe- lium in regions with low activity of B-hexosaminidase contained PAS-stained glycoconjugates (arrows, top panel), but staining was undetectable in regions with abundant enzyme activity such as those close to injection sites (arrows in lower panel). Bars: 2 mm (from top: first set of panels in a); 200 μm (insets in a); 100 μm (c, and from top: second, third, and fourth sets of panels in a); 50 μm (b).

Article Snippet: Immunohistological staining was performed with mouse monoclonal anti-N-acetyl-GM2 immunoglobulin M (MK1-16; Seikagaku, Tokyo, Japan; 1/100), rat anti-mouse CD68 (MCA1957; Serotec, Oxford, UK; 1/50), goat anti-GFAP (N-18, Santa Cruz Biotechnology, Calne, UK; 1/50) as primary antibodies.

Techniques: Staining, Marker, Injection, Labeling, Activity Assay

Journal: PLoS ONE

Article Title: Cholesterol Diet Withdrawal Leads to an Initial Plaque Instability and Subsequent Regression of Accelerated Iliac Artery Atherosclerosis in Rabbits

doi: 10.1371/journal.pone.0077037

Figure Lengend Snippet: Panel (A) Representative images of hematoxylin and eosin stained sections of all groups (Scale bar = 500 µm). Panel (B) Representative images of Oil Red O stained cryostat sections of all groups (Scale bar = 50 µm). Panel (C) Immunohistochemical staining showing CD68 positive cells in respective groups. (Scale bar = 50 µm) (D) Quantitative analysis of percentage lipid area (lumen area included) in respective groups (E) Quantitative analysis of CD68 positive labelling in respective groups. **p<0.01 and ***p<0.001 vs normal; ## p<0.01 and ### p<0.001 vs Baseline. (* indicates lumen, arrow heads indicate positive staining)

Article Snippet: Paraffin-embedded consecutive sections were stained with the following antibodies: Mouse monoclonal anti rabbit CD68 (RAM11, Dako, USA), alpha smooth muscle actin (1A4, Sigma, USA) and MMP-9 (56-2A4, Calbiochem, USA).

Techniques: Staining, Immunohistochemical staining

Journal: PLoS ONE

Article Title: Imaging Neuroinflammation In Vivo in a Neuropathic Pain Rat Model with Near-Infrared Fluorescence and 19 F Magnetic Resonance

doi: 10.1371/journal.pone.0090589

Figure Lengend Snippet: A , B show the affected right sciatic nerve from the CCI leg shown in stained with anti-CD68 antibody to reveal the presence of macrophages infiltrating the nerve. C , D CD68 positive cells are not present in the left leg (contralateral to surgery ) of the same CCI animal. E , F show a nerve from a separate CCI animal that also exhibits infiltration of CD68 positive cells. The boxed area is enlarged to reveal the granular cytoplasm (black arrow) of the macrophages, indicative of the presence of the nanoemulsion. Sham surgical sciatic nerve and non-surgical control sciatic nerve do not exhibit any CD68 positive cells ( G , H ) and ( I , J ). The fluorescent images A, C, G, and I were all acquired at the same sitting with the exact same image acquisition parameters. Macrophages grown in cell culture take up the nanoemulsion, exhibited as particles evident by both confocal fluorescence emissions 700–850 nm of NIR label (DiR) ( K , M ) as well as transmitted light ( L , N ).

Article Snippet: In separate immunohistochemical experiments using comparable protocols, the recovered control, sham and CCI sciatic nerves were prepared for examination using mouse anti rat CD68 antibody (MCA341R, AbD Serotech, Raleigh, NC) and Alexa fluor 488 donkey anti mouse secondary antibody (A-21202, Invitrogen, Carlsbad, CA) to assess the presence of macrophages that infiltrate the nerve.

Techniques: Staining, Control, Cell Culture, Fluorescence