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Overview

SEM Pathology

Table of contents

1. Examination of common tissue samples

Staining properties of platinum blue

Staining properties of osmium tetroxide

2. Examination of renal biopsy samples

3. Advantages of LVSEMs and future tasks

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Staining properties of platinum blue　#000002

(1) Normal rat tongue tissue (light-microscope images, LVSEM images)

Figure No.000002a

Staining: Platinum-blue staining

Microscope: Light-microscope
Sample: Paraffin section

Magnification: ×50

Scale bar: 100 μm

Image description
Light microscope image of a paraffin section (on a glass slide) of rat tongue stained with platinum blue 3), 4).

Figure No.000002b

Staining: Platinum-blue staining

Microscope: LVSEM
Sample: Paraffin section

Accelerating voltage: 15 kV

Magnification: ×50
Scale bar: 100 μm

Image description
LVSEM image of the paraffin section (on a glass slide) depicted in Fig. 000002a 3), 4).

Characteristic findings

Platinum blue is unique in that it functions as both a “backscattered electron signal enhancer” and a “histologically specific stain.” Differences in staining intensity produce varied shades of blue under a light microscope, whereas they produce a light-dark contrast under a LVSEM. These characteristics of platinum blue facilitate the identification of different types of cells and tissues. Furthermore, unlike light microscopes, LVSEMs allow for the use of platinum blue–stained paraffin sections to capture three-dimensional high-magnification images for morphologic analysis 3), 4).

Observations

Animal tissue samples stained with platinum blue show histologically specific staining that results from differences in the staining intensity of various cellular and tissue components. Thus, samples are characterized by differing shades of blue under a light microscope (Fig. 000002a) and as three-dimensional structures exhibiting light-dark contrast under a LVSEM (Fig. 000002b). These features facilitate the characterization of cells and tissues. In addition, when using a LVSEM, the scale of magnification can be increased up to several tens of thousands. This enables a more detailed analysis of the three-dimensional structures of cells and tissues. Stained images of cells and tissues can be roughly classified as follows:

【Staining properties of platinum blue】

  • Strongly stained (light microscope, dark blue; LVSEM, brightest)
    Examples: mast cells, epithelial tissue (stratified squamous epithelium, vascular endothelium)
  • Moderately stained (light microscope, light blue; LVSEM, bright)
    Examples: muscle tissue (striated muscle, smooth muscle), nerve tissue
  • Weakly stained or unstained (light microscope, bluish-white or colorless; LVSEM, dark)
    Examples: fibrous connective tissue (collagen fiber, elastic fiber)

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