Since high electron density tracers can easily diffuse in the interstitial space of cells, and tracers can also enter into cells if cell damage occurs, this method can be utilized to observe cell connections and cell damage. Commonly used tracers include lanthanum and peroxidase. The incubation method is generally used, in which the tissue block is incubated in the tracer solution. Some people also use vascular perfusion method, but the basement membrane can prevent the tracer from entering the cell gap or intracellular, so generally only when the study of vascular permeability changes in the vascular perfusion method, most of the cases with the incubation method. Source: Experimental Techniques and Applications of Immunohistochemistry

Operation method

basic program

Materials and Instruments

Tissue samples
NaOH glutaraldehyde lanthanum nitrate osmium-dimethylarsenate sodium buffer

Move

1. 4% lanthanum nitrate, pH 7.8, adjusted with NaOH, stirring while adding, so that the solution is milky white.

2. 1~1.5% lanthanum nitrate, 2%~3% glutaraldehyde-0.1 mol/L sodium dimethylarsinate buffer was used to fix the tissues before fixation for 2~4 h at 15~25℃. 3. 1% lanthanum nitrate-sodium dimethylarsinate buffer was rinsed.

3. 1 % lanthanum nitrate-sodium dimethylarsinate buffer was rinsed for 30 min.

4. 1 % lanthanum nitrate and 1 % osmium dioxide-sodium dimethylarsinate buffer fixation for 1-2 h. 5. Embedding and sectioning were performed as described above.

5. Embedding and sectioning as usual.

Caveat

1. Due to the limited penetration of the tracer, care needs to be taken not to go too deep when repairing the blocks.

2. The penetration of tracer is related to the incubation conditions, such as temperature, time, etc., so the actual application needs to be explored according to the specific situation.

3. Since lanthanum has an electronic staining effect, so in general there is no need to re-stain, of course, according to the routine method of staining can also be, and does not affect the results.

4. The time of tissue or cell in the liquid containing _ should not be too long, otherwise the section is difficult.

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