Question

1. A) Why is it important to control illumination for live cells? B) Briefly explain how live cell imaging chambers can be used to control four parameters (name and indicate importance) that contribute to maintaining healthy cells.

Answer 1

1. A) Living cells are stidied by live cell imaging, it is very very iportant tool to understand the cellular processes in real time. In live cell imaging high illumination or fluorescent labelling is used, both method cause phototoxicity or photobleaching. At high light intensity sensitive fluorophore get damaged due to phototoxicity or photobleaching and image quality is reduced. In in vivo conditions, many cells and tissue not exposed to high levels of light. High intensity of light can cause DNA damage and photobleaching due to rise in temperature of cells. Used fluorophore produced free raicals under high light intensity, that can destruct the cellular coponents. Thatswhy, control of illumination is important for live cells during live cell imaging.

B) Live cell iaging chambers control physiological conditions to aintaining cell health under microscope. There are four paraeters that control inside th live cell iaging chambers

1. Temperature: Cellular activity is very sensitive to temperature changes, change in temperature can affect cell adhesion, expression of protein, cell proliferation etc.

2. Oxygen level: As we know oxygen level is generally 21%, but the O2 levels tissues, and blood, are much lower. Generally in our body systems there are Variations in the oxygen concentration is comon e.g., 14% O2 is presentin the alveoli whereas 5% in peripheral tissues. during live cll imaging, control of oxygen level is an important parameter.

3. Carbon dioxide level and pH of medium: most cell has its optimum pH, carbon dioxide level influence the pH of the cell. Constant carbon dioxide level are important for for live cell iaging.

4. Humidityand evaporation: Concentration of different salts, nutrients etc., are crucial for cell existence. Any evaporation from the cell increases the substance concentration in the cell. Due to this any vital functions of living cells, affect that leads to impaired cell behavior e.g., altered proliferation, up- and downregulation of gene expression, and induction of apoptosis/cell death.