Material and Instrument:
Glass Slides
Cover Slip
Invereted microscope with phase plate.

Observation: Take out the cells from the cell culture plate by trypsinization or by 0.5% EDTA in PBS. Place a small amount of cells on the glass slide and cover them with cover slip. Observe the cells under the 40x objective using inverted microscope with phase plate. A typical observation of healthy and abnormal macrophage is given in the Figure 32.2.

Figure 32.2 Observation of macrophage in the inverted phase microscope phase.

Conclusions:  In a Typical observation, healthy cells will diffract the light rays aand as a result cell membrane, nucleus and cytosol can be observed. Where as in the case of disease or damged cells will show condensed nuclear content, several cell bodies or apoptotic bodies and scrambled membrane. The contrast of cytosol will not be very clear in the cells exposed to the cyto-toxic compounds.

Lab Experiment 31.3 : Determine the number of viable cell present in the cell culture using Trypan Blue exclusion method.
Material and Equipment

1. Glass Slides
2. Cover Slip
3. Invereted microscope with phase plate.
4. Hemocytometer
Trypan Blue solution.