Immunosero: Experiment # 1 - Preparation of Red Cell Suspension (Pg. 13-14)
Immunosero: Experiment # 1 - Preparation of Red Cell Suspension (Pg. 13-14)
Immunosero: Experiment # 1 - Preparation of Red Cell Suspension (Pg. 13-14)
204.
205. What is the purpose of preparing a red cell suspension?
Red cell suspension is a standard reagent used in many serological procedures. Red cell
suspensions have the required serum to cell ratio to allow the results of tests to be interpreted.
206. Is it permissible to make an approximation for the volume of NSS to be added to the
washed red cells? Justify your answer.
To avoid getting incorrect effects, one must use the precise volume or measurement of the NSS
to be applied to the washed red cells. In order for the red cell suspension to not be too heavy or
too diluted, it must be diluted properly. In order to avoid the phenomena or problems that are the
prozone and postzone phenomenon in each serological test and in blood banking, it is
necessary to do so. The equivalence would be achieved if correctly diluted and an ideal
antigen-antibody reaction will occur.
207. Calculate the volume of NSS to be added to a packed cell volume of 0.8mL to produce a
2% red cell suspension
Experiment # 2 – Serial Dilution (pg.19-20)
208.
209. Cite the differences between tube dilution and microtiter plate dilution
210. If you have a 1:10 dilution of the patient’s serum, how many parts of the sample and that of
the diluting fluid are present in the solution?
The solution will have 9 parts of the diluting fluid and 1 part of the sample.
212.
Hank’s Balanced Salt Solution - This is an isotonic solution that functions in regulating the
osmolality and pH and is used to maintain, for a short period of time, the cells found outside the
growth medium
Ficoll-Hypaque Solution - This is a density gradient solution used in the purification of
lymphocytes in high yield and purity from small and large volumes of human peripheral blood
RPMI & McCoy Medium - This medium is used to support the growth of a variety of cultures. It
is also used in the preservation of peripheral blood mononuclear cells
10% Fetal Calf Serum - This serum has a functions to supplement for basal growth medium
during cell culture
Swing-out Bucket Rotor Without Brake - This enables to adjust the angle of the tube when
performing a run.
Nylon Wool - This functions as a solid phase matrix and removes monocytes and B cells
213. Why is it important to isolate the peripheral blood mononuclear cells completely from the
other cellular components of blood?
A peripheral blood mononuclear cell (PBMC) is any blood cell having a round nucleus such as
lymphocyte, monocyte, or a macrophage. These blood cells are a critical component in the
immune system to fight infection and adapt to intruders. Peripheral blood mononuclear cells
(PBMC) give selective responses to the immune system and are the major cells in the human
body immunity. They contain several types of cells such as lymphocytes,monocytes or
macrophages. Because peripheral blood is the place where exposure to chemicals occurs,
these fundamentally important PBMCsare prone to be influenced by drugs and chemicals. This
is why the availability of PBMCs from peripheral blood is very important for researchers studying
toxicity of new drugs or chemical compounds.
214. What laboratory assays can be performed to assess the efficiency of the sample
preparation and isolation of the PBMC?
Trypan Blue Dye Exclusion Test: this is used to assess the number of cells that are still
viable in a cell suspension. Dead cells would turn blue, while living cells appear
clear/transparent when viewed microscopically.
217. Why should EDTA not be used in collecting blood specimens for an NBT test?
EDTA tubes should not be used because the anticoagulant can interfere with the results. It
inhibits phagocytosis thus resulting in results that are falsely decreased. The preferred
anticoagulant for NBT test is heparin.
218. Briefly discuss chronic granulomatous disease (CGD) and how it is diagnosed.
Chronic granulomatous disease (CGD) is a genetic disorder in which white blood cells called
phagocytes are unable to kill certain types of bacteria and fungi.