TRANSPORT PROCESS EXPERIMENT 2 RED BLOOD CELL FRAGILITY

1. Tabulate observation on the different dilution as to the: a. Supernatant b. Button of cells at the bottom of the tube

Tube No. 1 2 3 4 5 6 7 8 9 10

Supernatant Semi cloudy golden color Clear golden color Light, Golden and cloudy Clear golden clear Light golden clear Clear w/ golden traces @bottom Clear but partially cloudy Clear Clear Partially cloudy but clear

Button of cells @ bottom of the tube Less than half at bottom is compacted Compacted half, semi compact other half Compact at middle, hazy outer part Partially compacted button Partially compacted button Almost completely compacted Compacted but not fully solid Solid and compact cell button Solid and compact but darker shade of red Solid and Compact

2. Discuss and interpret results on the concepts of osmolarity and osmolality of the different dilution. In the experiment, red blood cells were placed in varying concentrations of NaCl. In test tubes 1-5, the concentration of NaCl reached. The cells swell but the water that crossed the membrane was not enough to lyse the cell. After centrifugation, the swelled red blood cells were collected and the supernatant remained clear. In test tubes 6-10, red blood cells were placed in NaCl solution with lower concentration, . Water rapidly enters the cells by osmosis and causes the cells to burst, a phenomenon known as hemolysis. Because there is a higher concentration of water outside the cell, water enters the cell by osmosis. In this case too much water enters and the cell swells to the point of bursting open. In the end pieces of cell membrane are left in the water

3. What is osmolarity of the different dilutions? And how is it calculated?

#1 V NaCl #2 V NaCl #3 V NaCl #4 V NaCl #5 V NaCl #6 V NaCl #7 V NaCl #8 V NaCl #9 V NaCl #10 V NaCl

5.0 NaCl 4.5 NaCl 4.0 NaCl 3.5 NaCl 3.0 NaCl 2.5 NaCl 2.0 NaCl 1.5 NaCl 1.0 NaCl 0.5 NaCl

5 x 1.8 29.225 4.5 x 1.8 29.225 4 x 1.8 29.225 3.5 x 1.8 29.225 3 x 1.8 29.225 2.5 x 1.8 29.225 2.0 x 1.8 29.225 1.5 x 1.8 29.225 1 x 1.8 29.225 .5 x 1.8 29.225

0.3 milliosmole/L 0.2 milliosmole/L 0.25 milliosmole/L 0.22 milliosmole/L 0.18 milliosmole/L 0.15 milliosmole/L 0.12 milliosmole/L 0.09 milliosmole/L 0.06 milliosmole/L 0.03 milliosmole/L

4. What is osmosis? What is the driving force of osmosis?

Osmosis is the flow of water between two solutions separated by a semipermeable

Difference in solute concentration. The driving force for osmosis is a difference in osmotic pressure caused by the presence of solute. Initially, it may be surprising that the presence of solute can cause a pressure, which is explained as follows. Solute particles in a solution interact with pores in the membrane and, in so doing. Lower the hydrostatic pressure off the solution. The higher the solute concentration,. The higher the osmotic pressure and the lower the hydrostatic pressure. Thus , if two solutions have different solute concentrations, then their osmotic pressures and hydrostatic pressures are also different; the difference in pressure causes water flow across the membrane. 5. Discuss the differences in the solutions and its effect on the RBC Isotonic solutions have the same effective osmotic pressure. When isotonic solutions are placed on either side of a semipermeable membrane, there is no difference in

membrane caused by a

effective osmotic pressure across the membrane, no driving force for osmosis and no water flow. In which hypertonic (higher effective osmotic pressure) and hypotonic(lower effective osmotic pressure) have different effective pressures. If these solutions are placed on either side of a semi permeable membrane, then an osmotic pressure difference is present. Thus, water flows from the hypotonic solution into the hypertonic solution because osmotic pressure difference is the driving force for water flow. -Isotonic solution 0.9 % NaCl -hypotonic solution- <.9% NaCl -the effect on RBC RBC burst in hypotonic (< 0.9 % NaCl), and shrink (crenate) in hypertonic solutions (> 0.9 % NaCl) 6. What is RBC fragility test? What diseases are associated with decreased and increased RBC fragility test? RBC fragility test a. it is a test that measures the resistance to hemolysis of red blood cells (RBC) exposed to hypotonic solutions b. RBC are exposed to a series of saline (NaCl) solutions with increasing dilution c. The sooner hemolysis occurs, the greater is osmotic fragility of RBC NORMAL RANGE: - hemolysis onset at: 0.45-0.5 % NaCl - hemolysis complete at: 0.3-0.33 % NaCl FACTORS AFFECTING OSMOTIC FRAGILITY - cell membrane permeability - surface-to-volume ratio INCREASED OSMOTIC FRAGILITY Hereditary spherocytosis Acquired spherocytosis DECREASED OSMOTIC FRAGILITY Thalassemia Sickle cell anemia Iron deficiency anemia
Spherocytosis is a disorder characterized by a defective RBC membrane and decreased surface-to-volume ratio. Characteristic round cells (spherocytes) are seen in blood smear and they are more fragile and break open in less hypotonic solutions than normal red blood cells