Heavy metal staining of paraffin epoxy and glycol methacrylate embedded biological tissue for scanning electron microscope histology

Details

• Personal Author:
  De Nee PB ; Frederickson RG ; Pope RS
• Description:
  This paper describes three techniques for preparing biological tissue sections for correlation of light microscopy (LM) and scanning (SEM) and transmission (TEM) electron microscopy. Tissues were embedded in paraffin, epoxy (EponAraldite), and butoxyethanol-glycol methacrylate (BGMA). Paraffin sections were stained with silver. Epoxy embedded tissue was postfixed in Os04, mounted on carbon-coated nylon grids and stained with uranyl acetate and lead citrate. BGMA sections were stained with uranyl acetate, lead citrate, silver proteinate and phosphotungstic acid. Epoxy and BGMA sections on polished carbon planchets were stained by flooding the surface of the planchet. Paraffin and BGMA sections were histologically stained for LM. Sections were viewed in the SEM using transmitted (TSEM), backscattered (BSE) and secondary (SE) electron imaging. Summation of BSE and SE signals was also used. SE imaging of deparaffinized sections shows tissue surface, while BSE imaging reveals deeper internal structure. Osmicated tissue shows good intracellular detail with SEM. BSE and SE images of BGMA and epoxy sections resemble LM and TEM images of alternate sections. TSEM images of epoxy sections provide good definition of thin extra-cellular fibrils. Large BGMA or many small epoxy sections can be stained and examined on a single planchet. The following application is recommended for each technique. Paraffin embedding - for the best correlation of surface topography and internal structure. Epoxy embedding - for the best intracellular details and tissue patterns, without interference from grid bars. BGMA embedding - for the best correlation of LM, SEM and TEM. [Description provided by NIOSH]
• Subjects:
  Histology Microbiology Occupational Health Pathology Safety
• Keywords:
  Analytical Processes Araldite Author Keywords: Backscattered Electron Imaging Biological Tissue Epon Glycol Methacrylate Heavy Metal Staining Microscopic Analysis Microscopy Paraffin Scanning Electron Microscopy

  More +
• ISBN:
  9780915802128
• Publisher:
  IIT Research Institute
• Document Type:
  Text
• Genre:
  Proceedings
• Place as Subject:
  OSHA Region 3 ; West Virginia
• CIO:
  National Institute for Occupational Safety and Health (NIOSH)
• Division:
  ALOSH - Appalachian Laboratory for Occupational Safety and Health
• Topic:
  Workplace Safety & Health
• Location:
  North America
• Pages in Document:
  83-92
• NIOSHTIC Number:
  nn:20047682
• Citation:
  Scanning electron microscopy: proceedings of the workshops on advances in biomedical applications of the SEM & STEM, March 31-April 1, 1977, Chicago, Illinois. Johari O, Becker RP, eds. Chicago, IL: IIT Research Institute, 1977 Mar; :83-92
• Editor(s):
  Johari O; Becker RP
• Federal Fiscal Year:
  1977
• Peer Reviewed:
  False
• Source Full Name:
  Scanning electron microscopy: proceedings of the workshops on advances in biomedical applications of the SEM & STEM, March 31-April 1, 1977, Chicago, Illinois
• Collection(s):
  National Institute for Occupational Safety and Health
• Main Document Checksum:
  urn:sha-512:7e60207a835af4e3d1d97ac51c054ca2cc0f04f56e1b19b9af84bbf18cff48a8b496ef4235854044e05cf520aee8fe3d771605a2fbd0234cff32901a628b9213
• Download URL:
  https://stacks.cdc.gov/view/cdc/202685/cdc_202685_DS1.pdf
• File Type:
  [PDF - 5.54 MB ]