By Kevin Vaughn
Immunocytochemistry of plant cells is the 1st booklet solely devoted to this subject. the 1st and biggest component of the publication is anxious with a bunch of confirmed protocols and diversifications on those protocols that may end up necessary, many constructed or transformed within the author's laboratory. the second one component of the publication covers the reports which have been released formerly on all the plant organelles. a variety of state-of-the-art micrographs from researchers worldwide are incorporated to illustrate common results.
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Additional resources for Immunocytochemistry of Plant Cells
Drops of lead citrate are transferred to Parafilm strips that are placed in Petri dishes in which sodium hydroxide pellets are placed in the corner. The sodium hydroxide keeps the concentration of carbon dioxide down during the staining process. Caution should also be exercised in exhaling while the sections are being post-stained. I even make the precaution of having no one around (and locking the doors) when I’m staining grids as I’ve ruined specimens when a well meaning colleague decides this is a good time for a conversation!
These numbers also substantiate that the micrographs presented in the paper represent a realistic representative of how good the labeling was. We have utilized a rather simple system for quantifying the gold particles. Our Zeiss microscope is centered at a 500/500 mark on the column entry mechanism. To ensure random sampling, we chose to only use the grid square nearest the center of this 500/500 entry point, or for low density labeling, all four grid squares around this 500/500 point. Take micrographs of all of the structures that are labeled and also areas of plastic, cell wall or starch grain where non-specific antibody sticking may be observed.
Keep the anti-contamination devices on not only to prevent contamination of the sample but in case protein and buffer particles are not washed cleanly from the sections. Because the beam tends to mark the areas of the section that have been examined, be certain that fairly low magnification micrographs are obtained before more high resolution micrographs are taken. g. ) For 15 nm gold, magnifications 12,5009 and greater in magnification are documenting the distribution of the gold particles. Besides these micrographs showing specific distributions, I also take micrographs over walls, starch grains and areas of plastic not containing specimen to document any background labeling.