Aqueous mounting media

AEC is soluble in alcohol and clearing agents, and must be mounted in aqueous mounting media. All other substrates are not soluble in alcohol or clearing agents. They may be dehydrated, cleared, and permanently mounted. These substrates can be used as single labels or to introduce multiple colors in a tissue section as shown for instance in Fig. 6.1 (see Chap. 7). 

Aqueous mounting media for Aqueous mounting media for Organic mounting media brightfield microscopy fluorescence microscopy... 

Ready to use adhesive aqueous mounting media are available from Immunotech, Dako, Vector Laboratories, Molecular Probes, etc. 

Antifade agents (free-radical scavengers) in aqueous mounting media prevent photobleaching of fluorophore labels under exposure to excitation light. 

This can be used in its acid form or as the sodium salt. The chromogens Fast Red TR and Fast Blue BB produce a bright red or blue end product, respectively. Both are soluble in alcoholic and other organic solvents, so aqueous mounting media must be used. Fast Red TR is preferred when staining cell smears. 

Aqueous mounting media - a solution miscible with buffers, does not need dehydration, and is used with fluorescence immunocytochemistry. 

Mounting media any available permanent resinous medium for DAB/hematoxylin-stained preparations fluorescence-free aqueous mounting media for fluorescence or confocal laser microscopy (e.g.. Fluorescent Mounting Medium, Dako). 

To each slide, apply one or two drops of non-aqueous mounting medium to the uncovered tissue and apply the coverslip. Apply the coverslip at an inclined angle to the slide, and lower it gently to avoid trapping air bubbles. 

Add one or two drops of aqueous mounting medium to the stained tissue and apply the coverslip. 

Rinse in distilled water and mount with an aqueous mounting medium, or proceed further with steps 5, 6 and 7. 

Add an aqueous mounting medium or an antifade solution, mount a coverslip and analyze using fluorescent microscopy with a fluorescein filter. 

The sections are washed in PBS for 5 min and incubated for 30 min in fluorescein isothiocyanate (FICT)-labeled swine antirabbit immunoglobulin conjugate at 1 20 dilution in PBS. After being washed in PBS for 5 min, the sections are mounted with an aqueous mounting medium. The sections are observed under an epiilluminating fluorescent microscope. The negative control is not incubated with the primary antibody, and a frozen section from a patient with the known condition is used as the positive control. 

AEG AEG (3-amino-9-ethylcarbazole) gives a red-brown alcohol soluble end-product. AEG requires aqueous mounting medium. 

Remove excess water aronnd the tissue section by touching edge of slide to a paper towel. Proceed below to step 22 for mounting in aqueous mounting medium or to step 23 for mounting in inorganic medium. 

Aqueous mounting medium (e.g., Immu-mount Shandon, Pittsburgh, PA). 

Optional) Mount the coverslip in Vectashield aqueous mounting medium and view the cells under fluorescence illumination to qualitatively assess cell loading of FDxLys. 

Mount with xylene-based mounting medium, like Canada balsam or DPX however, aqueous medium can also be used (see Sect. 3.5). 

Upon oxidation, AEC forms a rose red end product which is alcohol soluble. Therefore, specimens processed with AEC must not be immersed in alcohol or alcoholic solutions (for example, Harris hematoxylin). Instead, an aqueous counterstain and mounting medium should be used. AEC is unfortunately susceptible to further oxidation and when exposed to excessive light will fade in intensity. Storage in the dark is therefore recommended. 

Produce permanent preparations by mounting in an aqueous-based medium, such as Immu-mount, or in a xylene-based mounting medium, such as Fluoromount, following dehydration through an ethanol senes (i.e., 5 min each in 10, 20, 30, 40, 50, 60, 70, 80, and 90% ethanol in distilled water followed by three 5-min steps in absolute ethanol see Note 4). 

Wash slides in distilled water (3x15 min) and then cover sections with a cover-slip using an aqueous-based mounting medium (see Note 4). 

Sections developed with permanent labeling methods (e.g., with DAB) can also be counterstained using, for instance, cresyl violet or aqueous toluidine blue to stain Nissl substance. For permanent stains, the usual mounting medium is one of low viscosity so it gives a thinner adhesive layer of appropriate optical properties (e.g., Permount ). For fluorescence, a corresponding typical fluorescence-free mounting medium is preferred (e.g., VECTASHIELD ). 

Mounting mount sections in aqueous medium or balsam for brightfield microscopy or in anti-fade medium for fluorescence microscopy (see Sect. 3.2.2). Notes. A11 incubations are at room temperature unless otherwise noted. Nuclear dyes (DAPI, Hoechst 33342 and Propidium Iodide) supplied as lyophilized solids are usually reconstituted in methanol. The stock solutions (5 mg/ml) are stable for many years when stored frozen at <—20°C and... 

The first systematic investigation of steric interactions in a liquid film sandwiched between two macroscopic liquid bodies was that of Andrews et al. [53], who applied an electric field across the film, leading to a compressional force on the film that consisted of glycerol mono-oleate chains in hydrocarbon oils. The films were formed in a 1 mm hole in a Fluon disk that was mounted between two bulk aqueous electrolyte solutions. Sonntag et al. [54] have studied aqueous films sandwiched between two hydrocarbon layers. The aqueous film containing the nonionic surfactant was formed between two approaching emulsion droplets that were projected in the dispersion medium at the ends of two capillaries. The film thickness and its change were determined by means of an interferometric method. 

Light microscopy aqneons monnting medium Aquamount Aqueous Mountant (Lemer Laboratories, Pittsburgh, PA) Crystal/Mount Permanent Aqueons Monnting Medinm (Biomeda Corp., Foster City, CA). 

---