Gelatin coated slide

NOTE Brain sections are at the level of bregma -3.8 mm of the rat brain atlas (paxinos and Watson 1982). 20-Micron thick coronal sections of frozen rat brain were cut, thaw-mounted onto gelatin-coated slides, and incubated with (a) 10 nM 3H-PCP... 

APES may interfere with silver salt solutions in intensification steps of immunogold silver-staining techniques therefore, for those applications, poly-L-lysine or gelatin-coated slides are preferable. 

Sections (4 xm thick) of formalin-fixed and paraffin-embedded tissues are mounted on gelatin-coated slides, which are placed in a beaker containing 1,000 ml of 0.2 M sodium... 

The sections are rinsed in the same buffer and incubated for 90 min in secondary antiserum (Jackson ImmunoResearch Laboratories, West Grove, PA), diluted 1 50 with PBX-5% normal horse serum. They are rinsed in 0.1 M sodium phosphate buffer and then incubated for 1 hr in peroxidase antiperoxidase (PAP), diluted 1 100 with PBX. The sections are rinsed three times for 5 min each in the same buffer followed by distilled water rinses. They are incubated for 10 min in 50 ml of 0.05 M imidazole/0.05 M cacodylate buffer (pH 7.2) containing 50 mg of DAB. This is followed by an additional 10 min incubation after adding 200 xl H202. All incubations are carried out under constant agitation. The sections are washed in distilled water, placed in the same buffer, mounted onto gelatin-coated slides, dried, dehydrated, and cover-slipped with Permount. Note that the ABC procedure can be used instead of the PAP procedure. 

Fairbaim, H. W., 1943a. Gelatin coated slides for refractive index mounts. Am. Mineralogist 28 396. -------, 1943b. Packing in ionic minerals. Bull. Geol. Soc. Am. 54 1305 ... 

Fig. 20.2 Chronic lithium and valproate robustly increase bcl-2 immunoreactive neurons in the frontal cortex. Male Wistar Kyoto rats were treated with either Li2C03, valproate or saline by twice daily i.p. injections for four weeks. Rats brains were cut at 30 pm serial sections were cut coronally through the anterior portion of the brain, mounted on gelatin-coated glass slides and were stained with thio-nin. The sections of the second and third sets were incubated free-floating for 3 d at 4°C in 0.01 M PBS containing a polyclonal antibody against bcl-2 (N-19, Santa Cruz Biotechnology,...

After the completion of the in vivo microdialysis experiment, rats are euthanized with an overdose of sodium pentobarbital (100 mg/kg) and perfused intracardially with 0.9% saline followed by 10% formalin. The brains are extracted and stored in a 10% formalin solution until they are sectioned into slices (40-60 mm) consecutively through the guide cannula tract. The sections are mounted onto gelatin-coated glass slides and stained with thionin. An observer unaware of the rat s treatment or results verifies the cannula location for each rat. 

Tissue specimens are fixed with 4% paraformaldehyde and embedded in paraffin at 60°C for 1 hr. Sections (4 pm thick) are mounted on gelatin-coated glass slides, deparaffinized, and rehydrated in distilled water. They are treated with 0.005% pepsin for 15 min at 37°C, followed by heating in 0.01 M citrate buffer (pH 6.0) in a microwave oven (300 W) at 80°C for 15 min. The sections are washed in distilled water for 5 min, rinsed in 0.01 M PBS (pH 7.2) for 15 min, and treated with 0.3-1% H202 to quench endogenous peroxidase activity. 

Tissue sections of 5 pm thickness. Paraffin sections mounted on commercially available silane-coated glass slides, such as standard SuperFrost Plus (Fischer Scientific, Pittsburgh, PA) or on slides prepared using a home-made APES coating or frozen sections mounted on silanized, gelatin-coated or poly-L-lysine-coated slides (see Notes 1-4). 

Slide coatings reduce nonspecific DNA binding to glass, but are primarily used to prevent tissue loss. Gelatin coating, although excellent for frozen sections, does not work for wax-embedded sections... 

Pretreatment In this step, samples are placed on a gelatin-coated glass slide in order to ensure strong attachment. Bacterial cells may also be collected on filters (0.2 jim diameter). The bacterial cells are then dehydrated in an ethanol series. Lysozyme treatment is often applied particularly in the case of Gram-positive bacteria in order to perforate the peptidogly-con layer. Some permeabilization steps may involve treatments with 1 M hydrochloric acid (HCl). 

Excess gelatin is removed, and sections are dried on overnight at room temperature. If trypsinization is necessary to expose the antigen, then sections should be mounted on Biobond- or Vectabond-coated slides. For ISH, sections are also mounted on Biobond or Vectabond coated shdes, but additionally, following removal of wax in 100% IMS, the slides are removed and dried for 30 min at 37°C (9) prior to equilibration in PBS (see Notes 1-6). 

Sections do not adhere to slide. Sections dried onto sUde too briefly—dry sections overnight. Sections not mounted on either an adhesive solution or a coated slide— check mounting solution is 1% gelatin or that slides are coated as intended. Proteinase is contaminating the dilnent—use fresh diluent (bacteria grow readily in warm diluent ). 

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