Haematology – portfolio

Haematology – The examination of blood
Blood is an important body fluid composed of cells bathed in a fluid matrix called plasma. In humans, cells normally constitute about 45% of the blood whilst the other 55% is represented by the plasma. The principal cell types are described as red (erythrocytes) and white (leucocytes of various types).
Plasma is a straw coloured liquid. It consists of 90% water in which various substances are in solution or suspension. These include plasma proteins such as serum albumin, serum globulin and fibrinogen; ions such as sodium, calcium, bicarbonate etc., vitamins, hormones and dissolved products of digestion and excretion.
The red blood cells contain the respiratory pigment haemoglobin, which in bulk give the cells their distinctive red colour.
The main functions of blood are :-
TRANSPORT – e.g. oxygen, carbon dioxide, glucose, hormones.
PROTECTION – e.g. leucocytes important in maintaining the immune response.
HOMEOSTASIS – e.g. provision of stable internal environment, maintaining a steady osmotic pressure, ionic concentration and pH.
Blood is one of the most important body fluids and other body fluids such as lymph, interstitial fluid, cerebrospinal fluid etc. are derived from it by processes of filtration and/or secretion.
In this practical we will be examining mammalian (horse) blood. It is useful to be able to analyse the constituents of blood in order to know the normal parameters and to detect any abnormalities indicative of physiological adaptations or disease.

EXPERIMENT 1. BLOOD SMEAR
This technique is used to examine the proportions and types of cell found in blood.

1.Take a clean slide and with a glass pasteur pipette place a verysmalldrop ofblood on one end of the slide.
2.Take another microscope slide, hold it at an angle of 45o(as shown in the diagramabove)in frontbut not touching the drop of blood bring the slide back towards the blood and touch the drop of blood with it. Then push the second slide along the firstslide in a direction away from the drop of blood to make a blood smear. (This willbedemonstrated to you).
3.Allow the smear to air dry for several minutes to allowthe cellsto stickto the slide.This may be speeded up by waving the slide in the air.
4.Fix the smear by placing the slide in a coplin jar containing methanol for 5min.
5.On a rack over the sink, flood the slide with Giemsa stain using a plastic pipette.Leave for 5min.
6.Rinse off the stain with tap water.
7.Allow smear to air dry and examine under a microscope at X 10 and at X 40 and under oil immersion at X 100 if required. A cover slip is not required.
8.Examine the blood smear that you have prepared and attempt to find examples ofthedifferenttypes of cells,that have been described toyou in the introduction to thispractical,in your own smear.
9.Make a drawingof a red blood cell and drawings ofthedifferent types of whiteblood cell that you find. You will needto includethesein your lab reportforthis practical.
10.Identify thecellsand briefly state their function,asdescribed in the introduction, and on the posters provided.
11.Notethesize andrelative abundanceof the different types of blood cells.Name five cell types and state their primary function in blood.Include these in your report.Describe what alterations you would expect to see in the blood smear if someone had a bacterial infection.
EXPERIMENT2. Isolating blood cell types by lymphoprep™
Isolation of lymphocytes by buoyant density centrifugation. This will be demonstrated within the practical class using a blood sample.
1. Make a drawing of the blood preparation after centrifugation, label the cell types and include in your lab report. Estimate the haematocrit of the sample using this method and include in your lab report.

Blood Practical report
If you choose to write up this practical for assessment you should follow the format below (written in third person and past tense).

Title of the experiment

BACKGROUND: include an overview of the topic area for investigation including references

AIM: describe the aim of the experiment

METHOD: In brief, describe what was performed to conduct the experiment

RESULTS: describe the results or outcome of the experiments conducted, include any relevant diagrams, graphs, drawings of what was observed and label appropriately so that the examiner can see you understand the underlying concepts.

For this practical you will need to include:
1) A labeled blood smear, label cell type, nucleus, membranes, try to show differences in cell sizes and abundance that you observed.
2) Describe the differences in cell sizes and abundance you would expect to see in a normal blood smear.
3)Name five cell types in human blood and state their primary function. Describe what alterations you would expect to see in the blood smear if someone had (i) Leukaemia and (ii) Anaemia.
4) Make a labelled drawing of the lymphoprep™ after centrifugation. Describe percentage red blood cells you would expect in a normal haematocrit.

CONCLUSIONS: describe the conclusions you can draw from the experiment conducted. If possible, show how this relates to the subject area in general and diagnostic monitoring.

ERROR: Identify any areas of the experimentation that could lead to error with regard to sample, reagents and outcome relating to diagnostic testing

REFERENCES: Include at least 5 references that are relevant to the introduction or conclusions drawn within this report. Ensure you have references to some primary material, published manuscripts within this selection. Use the Harvard referencing system.

Word Count: Max 3000 words (main body text and figure legends, does not include references).

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