Journal: Journal of visualized experiments : JoVE

Article Title: Whole-Mount Staining, Visualization, and Analysis of Fungiform, Circumvallate, and Palate Taste Buds

doi: 10.3791/62126

Figure Lengend Snippet: Materials

Article Snippet: Immunohistochemistry staining list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Wash the tissues with 0.1 M PB, 3 x 15 min. Place the tissues in 1 mL tubes with blocking solution (3% donkey serum, 0.5% non-ionic surfactant (see the ), 0.1 M PB) at 4 °C overnight. table ft1 table-wrap mode="anchored" t5 caption a7 Name Company Catalog Number Comments 2,2,2-Tribromoethanol ACROS Organics AC421430100 2-Methylbutane ACROS 126470025 AffiniPure Fab Fragment Donkey Anti-Rabbit IgG Jackson ImmunoResearch 711-007-003 15.5μL/mL Alexa Fluor® 647 AffiniPure Donkey Anti-Rat IgG Jackson Immuno Research 712-605-150 (1:500) AutoQuant X3 software Media Cybernetics Blunt End Forceps Fine Science Tools FST 91100-12 Click-iT™ Plus EdU Cell Proliferation Kit Molecular Probes C10637 Follow kit instructions Coverglass Marienfeld 107242 Cytokeratin-8 Developmental Studies Hybridoma Bank (DSHB), (RRID: AB_531826) Troma1 supernatant (1:50, store at 4°C) Dissection Scissors (coarse) Roboz RS-5619 Dissection Scissors (fine) Moria MC19B Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 ThermoFisher Scientific A21206 (1:500) Donkey anti-Rabbit, Alexa Fluor® 555 ThermoFisher Scientific A31572 (1:500) DyLight™ 405 AffiniPure Fab Fragment Bovine Anti-Goat IgG Jackson Immuno Research 805-477-008 (1:500) Fluoromount G Southern Biotech 0100-01 Glass slides Fisher Scientific (Superfrost Plus Miscroscope Slides) 12-550-15 Goat anti-Car4 R&D Systems AF2414 (1:500) Imaris Bitplane pixel-based image analysis software Neurolucida 360 + Explorer MBF Biosciences 3D vector based image analysis software Normal Donkey Serum Jackson Immuno Research 017-000-121 Normal Rabbit Serum Equitech-Bio, Inc SR30 Olympus FV1000 (multi-Argon laser with wavelengths 458, 488, 515 and additional HeNe lasers emitting 543 and 633) Paraformaldehyde EMD PX0055-3 4% in 0.1M PB Rabbit anti-dsRed Living Colors DsRed Polyclonal Antibody; Clontech Clontech Laboratories, Inc. (632496) 632496 (1:500) Rabbit anti-PLCβ2 Santa Cruz Biotechnology Cat# sc-206 (1:500) Sodium Phosphate Dibasic Anhydrous Fisher Scientific BP332-500 Sodium Phosphate Monobasic Fisher Scientific BP330-500 tert-Amyl alcohol Aldrich Chemical Company 8.06193 Tissue Molds Electron Microscopy Sciences 70180 Tissue-Tek® O.C.T.

Techniques: Software, Dissection, Plasmid Preparation, Electron Microscopy, Labeling