4mp macrofire digital camera

Inoculated foot tissue was harvested from mice infected with WT or SODB1/Δsodb1 L. major into 10% neutral buffered formalin. Tissue was processed, paraffin-embedded, and longitudinal tissue sections were stained with hematoxylin and eosin (H&E). H&E stained sections were scored for inflammation. The “extent of inflammation” was determined by the number of 100x fields required to cross the area of inflammation in a tilling fashion (each 100x field was assigned a value of 1). The “relative quantity” of inflammation was determined by examining the dermis and hypodermis tissues and estimating whether inflammatory cells filled 0%, 5%, 10%, 25%, or ≥50% (and assigned values of 0, 1, 2, 3, 4, 5) of the space between the normal connective tissue cells. The total inflammation was determined by multiplying the “extent of inflammation” by the “relative quantity” of inflammation. Histologic images were captured at the tissue site showing the highest degree of inflammation using an Olympus BX51 microscope equipped with a 4MP Macrofire digital camera (Optronics) and using the PictureFrame Application 2.3 (Optronics). All images were processed identically by Photoshop (Adobe Systems Inc., Mountain View, CA).

Fixed tissue sections were processed for hematoxylin and eosin (H&E) staining and immunofluorescence microscopy as described previously [19 (link)]. Additional histology sections were processed for Periodic Acid Schiff (PAS) [20 ] and Picrosirius Red using stains made in house (http://www.ihcworld.com/_protocols/special_stains/sirius_red.htm). Immunoreactivity for PLIN2 and ACTA2 was visualized using secondary antibodies conjugated with Alexafluor 488 or Alexafluor 594 (Life Technologies) at dilutions of 1:500 and 1:250, respectively. Nuclei were stained with 4’, 6-diamidino-2-phenylindole (DAPI; Sigma Chemical Co.). Antibody to PLIN2 was raised in rabbits as described [21 (link)] and antibody to ACTA2 was purchased from Epitomics-Abcam. Immunofluorescence images were captured on a Nikon Diaphot fluorescence microscope and digitally deconvolved using the No Neighbors algorithm (Slidebook) as described previously [19 (link)]. Histologic images were captured on an Olympus BX51 microscope equipped with a 4mp Macrofire digital camera (Optronics) using the PictureFrame Application 2.3 (Optronics). Cross-polarized light was also used to enhance visualization of Picrosirius Red stained images as previously described [22 ]. All images were cropped and assembled using Photoshop CS2 (Adobe Systems, Inc.).

Fixed tissue sections were processed for hematoxylin and eosin (H&E) staining as described previously [65 (link)]. Histologic images were captured on an Olympus BX51 microscope equipped with a 4mp Macrofire digital camera (Optronics, Tokyo, Japan) using the PictureFrame Application 2.3 (Optronics). All images were cropped and assembled using Photoshop CS2 (Adobe Systems, Inc., San Jose, CA, USA).