Glime, J. M. and Wagner, D. H. 2017. Laboratory Techniques: Slide Preparation and Stains. Chapt. 2-2. In: Glime, J. M. Bryophyte 2-2-1 Ecology. Volume 3. Methods. Ebook sponsored by Michigan Technological University and the International Association of Bryologists. Ebook last updated 3 August 2017 and available at <http://digitalcommons.mtu.edu/bryophyte-ecology/>. CHAPTER 2-2 LABORATORY TECHNIQUES: SLIDE PREPARATION AND STAINS Janice M. Glime and David H. Wagner TABLE OF CONTENTS Preparing the Specimen....................................................................................................................................... 2-2-3 Cleaning Bryophytes.................................................................................................................................... 2-2-3 Washing Machine................................................................................................................................. 2-2-3 Embroidery Hoop................................................................................................................................. 2-2-3 Wash Bottle........................................................................................................................................... 2-2-3 HCl........................................................................................................................................................ 2-2-4 Aquatic Bryophytes .............................................................................................................................. 2-2-4 Sorting the Plants......................................................................................................................................... 2-2-5 Wetting Agents ............................................................................................................................................ 2-2-5 Soap...................................................................................................................................................... 2-2-6 Agral 600.............................................................................................................................................. 2-2-6 Clearing Leaves........................................................................................................................................... 2-2-7 Lactic Acid............................................................................................................................................ 2-2-7 KOH or NaOH...................................................................................................................................... 2-2-7 Chloral Hydrate..................................................................................................................................... 2-2-7 Dehydration.................................................................................................................................................. 2-2-7 Stains............................................................................................................................................................ 2-2-8 Staining Stems...................................................................................................................................... 2-2-8 Triple Stains................................................................................................................................... 2-2-8 Kawai Stem Staining Techniques.................................................................................................. 2-2-9 Acid Fuchsin................................................................................................................................ 2-2-17 Aniline Blue................................................................................................................................. 2-2-18 Congo Red................................................................................................................................... 2-2-18 Eosin............................................................................................................................................ 2-2-19 Fast Green.................................................................................................................................... 2-2-19 Fuchsin......................................................................................................................................... 2-2-19 Gentian Violet (=Crystal Violet)................................................................................................. 2-2-19 Janus Green.................................................................................................................................. 2-2-19 Methyl Green...............................................................................................................................2-2-19 Leaves................................................................................................................................................. 2-2-19 I2KI .............................................................................................................................................. 2-2-19 KOH or NaOH............................................................................................................................. 2-2-20 Methylene Blue............................................................................................................................ 2-2-22 Safranin O / Fast Green............................................................................................................... 2-2-21 Sphagnum Stains................................................................................................................................. 2-2-22 Methylene Blue............................................................................................................................ 2-2-22 Crystal Violet/Gentian Violet...................................................................................................... 2-2-22 Toluidine Blue O......................................................................................................................... 2-2-22 Reproductive Structures...................................................................................................................... 2-2-23 Iron Haematoxylon / Fast Green.................................................................................................. 2-2-23 Bulbils and Spores.............................................................................................................................. 2-2-23 Fluorescence and Fluorescent Dyes............................................................................................. 2-2-23 2-2-2 Chapter 2-2: Lab Techniques: Slide Preparation and Stains Staining Liverwort Capsules.............................................................................................................. 2-2-24 pH Testing................................................................................................................................................. 2-2-26 Weak Alkali............................................................................................................................................... 2-2-28 Cleaning Up Stains.................................................................................................................................... 2-2-28 Leaf Removal and Making Slides..................................................................................................................... 2-2-28 Sectioning ......................................................................................................................................................... 2-2-29 Razor Blades.............................................................................................................................................. 2-2-29 Cutting Techniques.................................................................................................................................... 2-2-30 Wax Mounts....................................................................................................................................... 2-2-30 Cutting Block ..................................................................................................................................... 2-2-30 Pith Sandwich Cutting Tool ............................................................................................................... 2-2-31 Chopping Method...............................................................................................................................2-2-31 Roll and Chop..................................................................................................................................... 2-2-31 Modified Roll and Chop..................................................................................................................... 2-2-32 Dissecting Microscope Hand Sections............................................................................................... 2-2-32 Double Slide Sectioning Technique................................................................................................... 2-2-33 Cryostat Sections................................................................................................................................ 2-2-34 Stems and Small Leaves...........................................................................................................................