Transcription of Protocol 3: Immunofluorescence on Frozen Sections
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Protocol 3: Immunofluorescence on Frozen Sections
Protocol 3: Immunofluorescence on Frozen Sections Brigitte Arduini, version 1, 2015-Mar-23 Modified from Manipulating the Mouse Embryo 3rd edition (2003). Nagy, Gertsenstein, Vintersten and Behringer, ed. Materials Phosphate Buffered Saline (PBS, 1X) Paraformaldehyde (PFA, 4%) see Support Protocol 1 Microscope slides, pre-coated Optimal Cutting Temperature (OCT) Compound 30% sucrose in PBS Cryotome blades (disposable or permanent depending on cryostat) Reagent scoop or slotted spoon Aluminum Foil Conical tube or screw-cap vial Pencil Slide box Humidity box (covered tray with dowels fixed to the bottom) Staining jars/slide holders Cover glass no.
2. Using a flat -bottom lab marker, inverted test tube or other suitable object, make a cup -shaped foil mold. The mold should be only slightly larger in diameter and in height than the tissue sample 2. 3. Fill the foil mold partialyl with OCT compound. 4. Remove tissue from sucrose and drain excess solution. Place the tissue sample in the OCT
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