Tech Notes

Tech Notes for EM and LM Applications (aka “Kirsch Notes” by Electron Microscopy Sciences)

You would like to know how to make formvar coated grids or why acetonitrile is a good replacement for propylene oxide?

The answers to these and many other questions, technical tips as well as information
on products for LM and EM can be found now on our webpage in this new tech note section.

(Kindly supported by Stacie Kirsch, EMS)

- General Lab and Safety Tips
  
  Calculating Solutions and Dilutions (PDF)
  
  Providing Solutions: General Lab Safety (PDF)
  
  Latex Gloves - Allergy and Alternatives
  
  Increase Stability and Shelf Life of Chemicals
- General Tips and Protocols for Sample Preparation for Microscopy (LM/EM)
  
  Quick Guide: Processing of Biological Samples for EM (PDF)
  
  Processing Steps of Biological Samples for TEM (PDF)
  
  Preparation of Gelatin Adhesive
  
  Decalcification of Animal Tissue
  
  Paraffin and Plastic for Preparation of Bone Marrow Specimen
  
  Mounting of Specimens for SEM with the Common Glue Gun
  
  Coverslip Treatment Test
  
  Reference: Rapid Techniques for Specimen Preparation
- Light Microscopy and LM Specimen Handling
  
  Protocol: Köhler Alignment for Light Microscopes (PDF)
  
  Permanox Plastic - A Non Fluorescent Material
  
  Poly-L-Lysine Coating of Slides
- Tissue Staining for Histology (LM)
  
  DEPEX - An Improved Mounting Medium
  
  Purification of Alcian Blue
  
  Hematoxylin Staining Tips
  
  A Methylene Blue-Azure II Stain
  
  A Toluidine Blue/Basic Fuchsin Stain for GMA Embedded Tissues
- Buffering, Fixation and Dehydration of Tissues (LM/EM)
  
  Preparation of Karnovsky's Fixative (PDF)
  
  Providing Solutions: Buffers for Electron Microscopy (PDF)
  
  Providing Solutions: Dehydrants for Electron Microscopy (PDF)
  
  Providing Solutions: Fixatives for Electron Microscopy (PDF)
  
  Use of Paraformaldehyde, Glutaraldehyde and Acrolein Buffer
  
  Use of Acetonitrile
  
  Fixative Solution for Fixing Cornea
  
  Prevent Paraformaldehyde Static Jumps
  
  Surgical Gloves - A Safety Tip -for usage with Glutaraldehyde
- Staining of Tissues with Heavy Metals (EM)
  
  Preparation of Osmium Tetroxide OsO4 Solution from Crystalline Form (PDF)
  
  Stability of Special Staining Solutions
  
  Air-Tight Container for Small Amounts of Osmium Tetroxide
  
  Neutralization and Disposal of Osmium Tetroxide
- Post-Staining of Sections with Heavy Metals (EM)
  
  Preparation of Post Staining Solutions (PDF)
  
  Lead Staining
  
  Common Lead Stock Solutions
  
  Handling of Lead Solutions
  
  Stable Lead Staining Solution
- Resin Embedding of Samples (EM)
  
  Providing Solutions: TEM Embedding Resins (PDF)
  
  Providing Solutions: Epoxy and Acrylic Resins (PDF)
  
  Embedding Samples in Epoxy and Acrylic Resins (PDF)
  
  Mixing Epoxy Resins (PDF)
  
  Preparation of Embedding Mixtures
  
  Epoxy Resins: Problems, Causes, and Solutions
  
  Embed812 - an Epon-812 Replacement
  
  Low Viscosity Kit Formation with ERL 4221
  
  Use of Accelerators for Epoxy Resins
  
  The Use of Flexibilizers and Plasticizers
  
  The Use of DMP-30 (2,2-Dimethoxy Propane)
  
  DMP-30 and DMAE VS BDMA
  
  Quetol and the Addition of Water
  
  Embedding Resin Stain Permability
  
  Storage of Epoxy Resins
  
  Storage of Embedding Resin Mixtures
  
  Reference: Re-Embedding of Tissue Poorly Embedded
  
  Reference: Re-Embedding of Paraffin-Embedded Specimen Tissue in Resin
  
  Adherence of LR White Sections to Glass Slides for Silver Enhancement Immugold Labeling
  
  LR White Technical Tips - Distinguishing LR White from Tissue
  
  LR White Technical Tips - Adjusting pH
  
  LR White Technical Tips - Improved Polymerization of Acrylic Resins
  
  Reference: Preparation of Glycol Methacrylate
  
  Reference: Flat Embedding with LR-White
- Ultramicrotomy and Sectioning Tips (LM/EM)
  
  Block Trimming for TEM (PDF)
  
  Thick Sectioning for TEM using Leica UC7 (PDF)
  
  Thick Sectioning for TEM using RMC PowerTome (PDF)
  
  Thin Sectioning for TEM (PDF)
  
  Flattening Semithin Sections
  
  Avoiding Contamination of Ultra-Thin Sections
  
  Dental Wax to Make Trufs
  
  Reference: Cryo Sectioning and the Use of Diamond Knives
  
  Elimination of Wrinkles in Semi-Thin Sections
- TEM Grid and Section Handling Tips (EM)
  
  Grids: Shiny Side vs Dull Side?
  
  Grids: Removing a Charge from the Surface of Grids
  
  Reaction of Ni and Cu Grids
  
  Picking Up of Carbon Film
  
  Reference: Simple Method for Handling Grids
  
  Techniques for Retrieving Thin Sections (PDF)
  
  Section Pick-up with the Perfect Loop
  
  Improving the Adhesion of Sections Filmless Grids
  
  On-Grid Enhancement
  
  Removal Of Lead Citrate & Uranyl Acetate Precipitates on LR White Sections
  
  Cryo Preparation using C-flat™
- Mounting of SEM Samples (EM)
  
  Providing Solutions: Mouting of SEM Specimens (PDF)
  
  Mounting Powders, Granules and Fibers
- Critical Point Drying (EM)
  
  Critical Point Drying with the EMS 3100 (PDF)
  
  Critical Point Drying with the EMS Q850 (PDF)
  
  Processing of Biological Samples with HMDS for SEM (PDF)
- Cryofixation of Tissues (EM)
  
  Preservation of Phospholipids
  
  Cryo-Protectants
- Immunolabeling (LM/EM)
  
  ***UNDER CONSTRUCTION***
  
  Use of Polyethylene Glycol
  
  AURION BLOCK 1
  
  AURION BLOCK 2
- EM Imaging and Analysis
  
  Providing Solutions: TEM Imaging Issues and Solutions (PDF)
  
  Guidelines: Qualitative X-ray Analysis (PDF)
  
  Protocol: Developing TEM Imaging Films (PDF)