You are involved in a study investigating the effects of a p

You are involved in a study investigating the effects of a particular neurotoxin which when administered destroys specified nerve cells. You are now preparing brain tissue for histological examination. Discuss in detail the follow questions: What type of stain would you use to determine whether there had been any changes in the total cell number following administration of the| neurotoxin. What stain might you use to determine whether certain cells had been structurally altered rather than destroyed (e.g. changes in dendritic number, length ectc?) How you prepare your tissue in order to observe the finest cellular detail using light microscopy? Indicate the amount of detail you would expect to see.

Solution

a) The cell number is determined using  Neubauer haemocytometer using trypan blue. Trypan blue dye stains the dead cells as it is taken up by the dead cells. Live cells donot take up the dye.

b) Specific flouroscent markers and stains can be used to determine whether the cells changed the dendritic number or length. Example- CD80 only appears in mature DC and Human ILT7 (novel pDC marker, CD85g) is specific dendritic cells marker.

C) The four steps in tissue preparation.

http://www.bio.bg.ac.rs/materijali_korisnika/priprema-materijala-za-sm-2.pdf