Silanized slides

Experimental samples are mainly derived from tissue culture cells, laboratory animals, or human tissues collected from hospitals after surgical biopsies and autopsies. With human and animal tissue specimens, it is important to arrest metabolic processes within 5-10 min of collection in order to preserve mRNAs from degradation by internal enzymatic reactions (26,27). Most hospitals use 10% buffered formalin as a tissue fixative. Subsequently, each tissue slice is trapped in a paraffin block. Series of 4-5-pm-thick sections are cut and mounted on silanated slides. Formalin-fixed archival tissues have been successfully used in in situ PCR and in situ hybridization protocols (28-32). However, the procedure for RNA protection is not always followed. It is often difficult to alter or control the routine procedures of hospitals for the required protection of mRNAs in surgically removed human tissues. 

Place each section on a silanated slide by scooping the slide nnder the floating tissne section and lifting the slide with the attached tissue section up and out of the waterbath. 

Take 4 (minimum) to 16 (maximum) paraffin-embedded tissue sections mounted on the silanated slides, and place them into the 65 °C oven, and melt the paraffin wax overnight. 

The interaction on silanated slides is thought to be the result of a combination of electrostatic and hydrogen bonding. Aminosilane surfaces are positively charged at neutral pH. Printing under alkaline conditions shifts... 

Biopsy tissue specimens are fixed with formalin and embedded in paraffin. Sections (5 xm thick) are mounted on silanated slides, heated at 56°C for 1 hr, deparaffinized in xylene, rehydrated in graded ethanols, and rinsed in distilled water. They are placed in 0.01 M citrate buffer (pH 6.0) and heated in a microwave oven for six cycles of 5 min each, followed by cooling at room temperature for 20 min. Endogenous peroxidase activity is blocked with hydrogen peroxide in distilled water for 8 min, and nonspecific background staining is prevented by treatment with nonimmune horse serum for 20 min. 

Silanized Slides (Code S3003) or slides coated with other suitable adhesives... 

Excessive or unevenly applied subbing agent on poly-L-lysine, charged, or silanized slides. Some IHC reagents may bind to these products, resulting in a light stain over the entire slide surface. Some slides may be unevenly coated, and will exhibit the above problems on only a portion of the tissue or glass. 115-121... 

P 0425, Sigma amino silane CMT-GAPS slides 2549, Corning Silanated slides, CSA-25, TeleChem). 

Lung tissue, or other tissue with a thin, open architecture, may be cut on charged or silanized slides to prevent the tissue from non-specifically adhering to the polymer during microdissection. In general, coated slides are not used for microdissection because of the increased adhesive forces between the tissue and the slide. Effective microdissection is a balance between three adhesive forces ... 

Silane slide coating solution 2% (v/v) 3-aminopropyltriethoxysilane (Sigma, cat. no. A3648) in acetone. Prepare fresh just before use (see Section 3.2.). 

Dip for 5 s into freshly prepared silane slide coating solution. 

Prehybridization Method (Amino-Silane Slides Such as CMT GAPS)... 

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