A method for co-localization of tubular lysosomes and microtubules in macrophages: fluorescence microscopy of individual cells.
Routinely used procedures for chemical fixation often fail to preserve delicate membrane-bounded tubular structures in a variety of cell types. Fixation procedures commonly employed in immunocytochemical studies for localization of structural proteins, such as those found in cytoskeletal elements, may also degrade these tubular structures. Here we describe a procedure that preserves the elaborate tubular lysosome system found in stimulated macrophages and allows the subsequent immunofluorescence localization of microtubules in the same cells. Use of this methodology permits the assessment of the spatial relationship between tubular lysosomes and microtubules in macrophages.
Robinson
J M
JM
Department of Cell Biology, Neurobiology, and Anatomy, Ohio State University, Columbus 43210, USA.
Chiplonkar
J
J
Luo
Z
Z
D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects; D005404:Fixatives / Q000494:pharmacology; D005557:Formaldehyde / Q000494:pharmacology; D017110:Freeze Substitution / Q000379:methods*; D005976:Glutaral / Q000494:pharmacology; D007546:Isoquinolines; D008247:Lysosomes / Q000648:ultrastructure*; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects / Q000648:ultrastructure*; D000432:Methanol / Q000494:pharmacology; D051379:Mice; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence / Q000379:methods*; D008870:Microtubules / Q000648:ultrastructure*; D011108:Polymers / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D016707:Tissue Fixation / Q000379:methods*; D014404:Tubulin / Q000276:immunology
D005404:Fixatives; D007546:Isoquinolines; D011108:Polymers; D014404:Tubulin; D005557:Formaldehyde; C017475:lucifer yellow; D013755:Tetradecanoylphorbol Acetate; D005976:Glutaral; C003043:paraform; D000432:Methanol