3-3'DABminobenzidine (DAB) solution (Dako Corporation, CA, USA) was then added in order to obtain staining.
Sections of 95 nm were then obtained, stained with uranyl acetate and lead citrate, and viewed on a JEOL 1200 EX transmission electron microscope (JEOL USA Inc., Peabody, MA).
Splenocytes were obtained, stained and analyzed by FACS to evaluate lymphoid DCs (CD11c+ CD8α+), myeloid DCs (CD11c+ CD8α−), plasmasytoid DCs (CD11c+ B220+), macrophages (CD11c−, CD11b+), B cells (CD11c− B220+), and natural killer cells (NK 1.1+).
The interphalangeal joint sections were obtained, stained with eosin-haematoxylin stain and viewed under 100 X magnifications.
Two micrometer sections were obtained, stained with hematoxylin-eosin and observed with an Olympus BX-40 light microscope.
Forty-two (84.0%) of the 50 cases for which immunohistochemical data could be obtained, stained positively for pRb2 while eight (16.0%) were negative (Table 2).
Similar results were obtained staining the same area with Abs anti-LMP7 (data not shown) and anti-PA28-α (Fig. 1C, D).
Specificity of the immunofluorescent staining reaction was established by failure to obtain staining with normal serum, with smooth muscle antibody serum neutralized by homogenates of smooth muscle or extracts containing actin derived from smooth muscle.
According to the forensics team's written report, which the newspaper La Nación obtained, stains in the bathroom sink had been scrubbed away, and the position of the gun was inconsistent with Nisman's having shot himself.
The PFO and PT alternate sections were always processed together in order to allow comparison of the obtained staining.
Tissue samples were processed to obtain DAPI stained nuclei suspensions following the method of Otto et al. [ 19] with modifications.
