Glime, J. M. and Wagner, D. H. 2017. Laboratory Techniques: Slide Preparation and Stains. Chapt. 2-2a. In: Glime, J. M. Bryophyte 2-2a-1 Ecology. Volume 3. Methods. Ebook sponsored by Michigan Technological University and the International Association of Bryologists. Ebook last updated 14 April 2021 and available at <http://digitalcommons.mtu.edu/bryophyte-ecology/>. CHAPTER 2-2a LABORATORY TECHNIQUES: SLIDE PREPARATION AND STAINS Janice M. Glime and David H. Wagner TABLE OF CONTENTS Preparing the Specimen ..................................................................................................................................... 2-2a-3 Cleaning Bryophytes .................................................................................................................................. 2-2a-3 Washing Machine ............................................................................................................................... 2-2a-3 Embroidery Hoop ............................................................................................................................... 2-2a-3 Wash Bottle......................................................................................................................................... 2-2a-3 HCl ...................................................................................................................................................... 2-2a-4 Ultrasound ........................................................................................................................................... 2-2a-4 Aquatic Bryophytes ............................................................................................................................ 2-2a-4 Dealing with Old Specimens ...................................................................................................................... 2-2a-5 Sorting the Plants ....................................................................................................................................... 2-2a-5 Wetting Agents .......................................................................................................................................... 2-2a-5 Soap .................................................................................................................................................... 2-2a-6 Agral 600 ............................................................................................................................................ 2-2a-6 Rehydrating Capsules ................................................................................................................................ 2-2a-6 DulcoEase ........................................................................................................................................... 2-2a-6 Clearing Leaves ................................................................................................................................................. 2-2a-7 Lactic Acid ................................................................................................................................................. 2-2a-7 KOH or NaOH ........................................................................................................................................... 2-2a-7 Chloral Hydrate .......................................................................................................................................... 2-2a-7 Dehydration ....................................................................................................................................................... 2-2a-8 Stains ................................................................................................................................................................. 2-2a-8 Staining Stems ........................................................................................................................................... 2-2a-8 Triple Stains ........................................................................................................................................ 2-2a-8 Kawai Stem Staining Techniques ....................................................................................................... 2-2a-9 Acid Fuchsin ..................................................................................................................................... 2-2a-17 Aniline Blue ...................................................................................................................................... 2-2a-18 Congo Red ........................................................................................................................................ 2-2a-18 Eosin ................................................................................................................................................. 2-2a-19 Fast Green ......................................................................................................................................... 2-2a-19 Fuchsin .............................................................................................................................................. 2-2a-19 Gentian Violet (=Crystal Violet) ...................................................................................................... 2-2a-19 Janus Green ....................................................................................................................................... 2-2a-19 Methyl Green .................................................................................................................................... 2-2a-19 Leaves ...................................................................................................................................................... 2-2a-19 I2KI ................................................................................................................................................... 2-2a-19 KOH or NaOH .................................................................................................................................. 2-2a-20 Safranin O / Fast Green ..................................................................................................................... 2-2a-21 Sphagnum Stains ...................................................................................................................................... 2-2a-22 Methylene Blue ................................................................................................................................. 2-2a-22 Crystal Violet/Gentian Violet ........................................................................................................... 2-2a-22 Toluidine Blue O .............................................................................................................................. 2-2a-22 Reproductive Structures ........................................................................................................................... 2-2a-23 2-2a-2 Chapter 2-2a: Lab Techniques: Slide Preparation and Stains Iron Haematoxylon / Fast Green ....................................................................................................... 2-2a-23 Bulbils and Spores .................................................................................................................................... 2-2a-23 Fluorescence and Fluorescent Dyes .................................................................................................. 2-2a-23 Staining Liverwort Capsules ..................................................................................................................... 2-2a-24 pH Testing ................................................................................................................................................ 2-2a-26 Weak Alkali .............................................................................................................................................. 2-2a-28 Cleaning Up Stains ................................................................................................................................... 2-2a-28 Leaf Removal and Making Slides .................................................................................................................... 2-2a-28 Avoiding Air Bubbles ............................................................................................................................... 2-2a-29 Sectioning ........................................................................................................................................................ 2-2a-29 Razor Blades ............................................................................................................................................. 2-2a-30 Cutting Techniques ................................................................................................................................... 2-2a-30 Wax Mounts ...................................................................................................................................... 2-2a-30 Cutting Block .................................................................................................................................... 2-2a-31 Pith Sandwich Cutting Tool .............................................................................................................. 2-2a-31 Chopping Method .............................................................................................................................