2. Your IA has created a class tube of Tetrahymena that contains 500 microliters

Question

2. Your IA has created a class tube of Tetrahymena that contains 500 microliters of cells + 500 microliters of 1% India ink. You will sample the tubes at 0, 5, 10, 20 and 30 minute time points. To sample, micropipette 20 microliters of cells + ink suspension onto a microscope slide. UNDER THE FUME HOOD, add 5 microliters of 1% glutaraldehyde to the drop of cells and mix using pipet tip (eject micropipette into beaker in fume hood). Add coverslip.(2) What is the purpose of the fixative glutaraldehyde? Why is it important?

Explanation / Answer

Purpose of fixative glutaraldehyde:

Glutaraldehyde (Chemial formula: C5H8O2) is a cross linking agent and hence used as fixative. Fixation preserves the cells and tissues by inhibiting any reactions and maintains a replica of living system in vitro. Glutaraldehyde causes deformation in the alpha helix structures of proteins which yields a cross linked fixed product that can be clearly demonstrated and studied using microscope

Why Glutaraldehyde is important?

Glutaraldehyde fixing is important because it maintains the cytoplasmic and nuclear details intact in the sample which is useful for studying the cell. It can increase the strength and rigidity of the sample and helps in maintainence of shape and structure which can be used for precision study. Hence, it is widely used as a cross linking agent in histology, pathology, cell biology, immunochemistry etc. It is most ideal for the sample fixation intended for study under electron microscopy. It is also used in Enzyme immobilization.

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