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1. Presenter: Dr. Alina Khadka
1st
year Resident
BLOOD AND ITS COMPONENT
AND BLOOD BANKING

2. BLOOD
• Specialised connective tissue.
• Total volume :5-6 L (7-8% body weight).
• PH : 7.35- 7.45
• Component:
- Plasma (55%)
- Blood cells(45%)

3. PLASMA
• Water (90-92%)- solvent, medium of transport.
• 7-8 % protein: (albumin, globulin,fibrionogen)
-Albumin- oncotic pressure, carrier protein.
-Globulin- immunity (Ig)
-Fibrinogen- Clotting.
• Others :electrolytes, nutrients, hormones ,waste
product.

4. BLOOD CELLS
• Red blood cells (erythrocytes).
• White blood cells(leukocytes).
• Platelets(thrombocytes)

5. RED BLOOD CELLS
• Biconcave disc shaped , measuring about 7-8 micro
meter.
• Normal count:
Male-5±0.5million/microlitre
Female- 4.3±0.5 million / microlitre
• Life Span:120 Days
• Hemoglobin : O2 and Co2 transport
Male-15±2 g/dl
Female-13.5±1.5 g/dl

6. WHITE BLOOD CELLS
• Nucleated cells of blood.
• Count:4000-10000
• Classification:
1.Granulocytes
2.Agranulocytes

7. WBC
• Granulocytes: Neutrophils
Eosinophils
Basophils
• Agranulocytes: Monocytes
Lymphocytes

8. Neutrophils
• Account 40-60% of total WBC counts with
2- 7x10^9 / L
• Uniform in size about 12-14 micrometer in
diameter.
• Have a pink/orange cytoplasm that is due to
presence of fine granulation.
• Have a segmented nucleus usually three nuclear
segment (lobes) connected by tapering
chromatin strands.

9. EOSINOPHILS
• Account 1- 6 % 0f leukocytes.
• About 12-17 micrometer in diameter.
• Usually have 2 nuclear lobe and cytoplasm is
packed with distinctive spherical gold / orange
(eosinophilic ) granules

10. MONOCYTES
• Account 2-10 %
• Diameter: 15-18 micrometer.
• Largest circulating leucocytes.
• Cytoplasm: bluish grey
• Nucleus: Large, curved, horse shoe shape with
no segmentation
• Chromatin: fine , evenly distributed

11. Lymphocytes
• 20-40% of total WBC count.
• Count: 1-3 thousand / microlitre
• Small lymphocytes: small with a thin rim of cytoplasm,
occasionally containing scanty azurophilic granules.
Nucleus: 7-10 micrometer
Have high N:C ratio
• Large lymphocytes: Abundant pale blue cytoplasm
containing azurophilic granules. The N:C ratio is lower
and nucleus may be round oval or indented with less
condensed chromatin.

12. • Reactive lymphocytes
-have slightly large nuclei with more open
chromatin.
-Abundant cytoplasm that may be irregular.
-Seen in – Viral infection such as Infectious
mononucleosis.

13. BASOPHILS
• Rarest < 1% in circulating leucocytes.
• Nuclear segment tends to fold up on each other,
resulting in a compact irregular dense nucleus
resembling a closed lotus flower.
• Granules: Large variable size dark blue or purple
often obscure the nucleus.
• Granules are rich in histamine, serotonin and
heparin.
• Absolute basophil count: 20- 100

14. PLATELETS
• Are small non nucleated fragments of cytoplasm
released from megakaryocytes.
• Small, round or oval, biconvex cytoplasmic disc,
varing in size from 1.5 to 3.5 micrometer in
diameter.
• Normal count:280 + 130 x 10^9 /L
• Life span: 5 to 10 days
• Irregular in outline with fine red granules that
may be scattered or centralised.

15. Blood banking
• Blood bank: A blood bank is a center where blood is
gathered as a result of blood donation, is stored and
preserved for later use in blood transfusion.
• Typically refers to a division of a hospital where the
storage of blood products occur and where proper
testing is performed ( to reduce the risk of
transfusion related events..
•

16. Continue
Blood banking includes tasks related to:
1. Blood collection
2. Processing
3. Testing
4. Separation
5. Storage

17. Functions of Blood Banking
• Blood donor selection
• Donor pre counselling
• Donor blood collection
• Donor post counselling
• Blood tests
• Blood storage
• Blood transfusion

18. Blood Donor Selection
• To assess the suitability of an individual to be a blood donor so that
donation is safe for the donor and the blood products derived from
this donation are safe for the recipients.
• Process should be carried out in accordance with written standard
operating procedures:
1.WELL BEING
• The donor shall be in good health, mentally alert and physically fit
and shall not be inmates of jail or any other confinement.

19. 2. AGE
• Minimum age 18 years
• Maximum age 65 years
• First time donor shall not be above 60 years of age
• For repeat donor upper limit is 65 years
• For aphaeresis donor 18-60 years
3.WEIGHT
• 350ml- 45 kg
• Apheresis- 50 kg
4. DONATION INTERVAL
• For whole blood donation, once in 3 months (90days) for males
and 4 months (120days)
for females.

20. 5. BLOOD PRESSURE
• 100 -140mm Hg systolic; 60-90mm Hg diastolic with or without
medication
6.Pulse
• 60-100 beats per minute
7.Temperature
• Afebrile; 37 /98.4 F
8.Haemoglobin
• It should be above 12.5g/dl
9.Meal
• The donor shall not be fasting before the blood donation or
observing fast during the period of blood donation and last meal
should have been taken atleast 4 hrs prior to donation

21. 10. ALCOHOL
• Donor shall not have consumed alcohol and show signs of
intoxication before the blood donation.
• The donor shall not be a person having regular heavy alcohol intake
11. PREGNANCY AND RECENT DELIVERY
• Defer for 12 months after delivery
12.ABORTION
• Defer for 6 months after abortion
13.BREASTFEEDING
• Defer for period of lactation

22. 14.MENSTURATION
• Defer for the period of menstruation
15.NON SPECIFIC ILLNESS
16.SURGERY
17.RESPIRATORY INFECTIONS
18.CARDIOVASCULAR SYSTEM

23. DONOR PRE COUNSELLING
• Explain about donation process and benefits of
blood donation
• Ensure volunteer blood donation
• Encourage HIV test
• Develop safe donor pool

24. DONOR BLOOD COLLECTION- PROCESS
• Blood donation is carried out under the supervision of trained, skilled
technicians.
• The entire procedure, from start to finish, does not take more then 45 minutes
• The blood is usually drawn from the median cubital vein, from the inside of the
elbow
• An antiseptic such as iodine is used to clean the skin above this vein
• This helps to prevent bacterial infection at the site of puncture and also helps
to prevent
the blood drawn from being infected.
• A tourniquet may be used to elevate the blood pressure in the veins of the
arm.
• This helps to ease and speed up the process.
• Sometimes the donor is given an object to squeeze repeatedly in order to
increase blood flow to the target vein
• Invariably a needle with larger gauge is used to minimize the shearing forces
that can cause damage to the RBCs.

26. DONOR POST COUNSELLING
• Tell about donor blood details (test results)
• Explain about healthy lifestyle
• Explain about to take nutrition
• Encourage HIV test
• Encourage AIDS test

27. BLOOD TESTS
All donations must be tested for the following infectious disease and
found to be negative for antibodies to:
• Human immunodeficiency virus ( anti HIV-1/2)
• Hepatitis C virus ( anti- HCV)
• Hepatitis B core antigen ( anti- HBc)
• Non reactive for hepatitis B surface antigen (HBsAg)
• Hepatitis B virus
• Syphilis

28. Figure: Blood grouping Test

29. DONOR BLOOD STORAGE
Donated blood was stored in 5 different ways:
1.Whole blood
2.Packed RBC
3.Fresh frozen plasma
4.Platelet rich plasma
5.Cryoprecipitate

30. WHOLE BLOOD
• A unit of blood collected from a single donor in a sterile
container with appropriate anticoagulant( CPDA-1 ).
• Volume :450 ml of blood.
• 56ml of anticoagulant in collection bag
• Anticoagulant CPD- Citrate Phosphate Dextrose
• Shelf Life- 35 days at 2-6C
• Indication:- Massive transfusion
Exchange transfusion

31. Complications of donation: :
 Bruises, soreness and hematoma at venipuncture site.
 Nerve irritation and/or injury and arterial puncture(less common)
 Vasovagal reactions, with syncope
 Fatigue, nausea and vomiting
 Risk of volume overload in patients with chronic anaemia and
compromised cardiovascular function

32. Red blood cells preservation and storage
ANTICOAGULANT: CPDA-1
• CITRATE: Anticoagulation by binding of calcium in plasma.
• Phosphate: Acts as a buffer to minimize the effects of decreasing pH in blood.
•
• Dextrose: Maintenance of red cell membrane and metabolism.
• Adenine : Generation of ATP (energy source).
• Allows storage of RBC concentrates for 35 days.
• About 20% of anticoagulant containing plasma is left to provide metabolic
substrate for RBC during storage.
• Stored at 2-6 C to maintain optimum function.

33. Changes in Red cells during storage:
• STRUCTURAL CHANGE
 RBCs become spherical with surface
projections(spheroechinocytosis). Loss of membrane lipids
and proteins
• BIOCHEMICAL CHANGES:
 Decreased glycolysis
– Loss of potassium
– 2,3-DPG depletion

34. Blood components
• Blood components can be separated from one another by
centrifugation due to differences in their specific gravities whereas
blood derivatives use a chemical (eg. Ethanol fractionation) in
varying concentration and temperature for its separation.
• Separation is carried in double or triple bags(satellite bags)with
closed integral tubing.
• Blood should be processed for component separation within 6
hours of collection.
 Primary goal of component preparation is to provide right
component to the right patients in right time in right quality.

35. NEED FOR SEPARATION
• Optimal survival of each constituents.
-In whole blood stored at 2-6 C, platelets stay viable for 1
day and factor V and VII decreased.
-While after separation , platelets stay upto 5 days and
Factor V and VII can store as FFP for 1 year at -30C.
• Transfusion of only required components thus
avoids the use of unnecessary component which
could be contraindicated in a patients.
• One donor can save several patients.

36. Advantages of component therapy
• Maximization of yield of products from a single blood
donation.
• Ability to use the optimal products for specific diseases( thus
minimizing the amount of unnecessary foreign material the
recipient is exposed to).
• Improved quality and functional capacity of each components
when varied storage conditions and shelf lives were applied

37. Blood component types
Whole blood
Red cells
Packed RBCS
RBCS in
additive
solution
Leucocyte
reduced RBCS
RBCS ,
deglycerolized
Irradiated
RBCS
Washed
RBCS, Frozen
RBCs
Granulocyte
s, pheresis
Plasma
FFP, Thawed
plasma
Cryoprecipitate
Fractionated
products
Human albumin,
immunoglobulin, Factor
VIII, IX
Platelet
concentrates,
leucocyte reduced,
pheresis

38. Preparation of blood components
Equipments:
• Collection bag:
• Refrigerated centrifuge
• Plasma extractor
• Cooling water bath
• Freezer(-18degree centigrade or less)
• Blood refrigerator (1-6 degree centigrade)
• Platelet incubator

39. 1)PACKED RED CELLS
• Red cells have a higher
specific gravity than the
plasma , thus settle in
lower portion of the bag

40. PACKED RBC
• Red cells with 1/3 of the original plasma.
• contains mostly RBC, WBC and less amount of plasma.
• 45gm of hemoglobin per unit
• 250ml of blood stores
• Stored at 2-6C
• Shelf life: 35 days(CPDA1) Photo: showing extraction of
plasma and packed red cells from
whole blood

41. Indication:
-Symptomatic anemia
- Bleeding during surgery.
-Severe haemorrhage for patients with surgical
complication.
-Trauma
-Cancer
1 unit raises – 1.5 g/dl of Hb in an adult.

42. STEPS IN SEPARATION
1 • Store the bag at 2-6 °C till processed
2
• Place bag in buckets of refrigerated centrifuge
and balance the opposite bag
• Centrifuge at 4000 rpm (hard spin) for 10 min
at 2-6°C
3
• Express approx. 4/5th
of the plasma in the
satellite bag
• Keep red cell at 2-6°C and plasma at -18 °C or
below.

43. 2)Red cells in additive solution (Red cell
suspension)
• Commonly used additive solution is SAGM which contains ( saline, adenine, glucose,
and mannitol.
• Advantages of this method are :
 Maximum amount of plasma can be removed for preparation of plasma components.
 Red cells with improved viability are obtained (shelf life increases from 35 days to 42
days).
•
 Flow of infusion is improved due to reduction in viscosity.
 Indications for red cells in SAGM are similar to those for packed red cells.
 contraindicated for exchange transfusion in neonates

44. 1 •collection of whole blood in the primary collection bag (containing CPDA-1)
2 •maximum amount of plasma is removed (after centrifugation) and transferred to one satellite bag.
3 •The additive solution from the second satellite bag is transferred into the primary collection bag (containing packed red cells) in a closed circuit

46. Leucocyte poor/ depleted RBCS
• WBC typically contaminate both red cell and platelet components with the
concentration of approx. 10 ^9/ product
• leucocyte depleted red cells should contain < 5×10^6 white cells per bag.
• obtained by passing blood through a special leucocyte-depletion filter at the
time of transfusion
Advantage:
• Minimizes sensitization to HLA antigens (e.g. in patients with severe aplastic
anaemia who are likely to receive allogeneic bone marrow transplant).
• Prevention of Febrile non- haemolytic transfusion reaction.
• Reduces the risk of transmission of cytomegalovirus (CMV).
• However cannot prevent graft vs. host disease

47. WASHED RED CELL
• Packed red cells can be washed with normal saline using
automated cell washers.
• Capable of removing approximately 99% of plasma proteins
from red cell products.
ADVANTAGES:
• Remove incompatible plasma proteins, white cells, and
platelets.
• Prevent severe allergic reactions (which are thought to be
triggered by donor plasma protein.
• Reduce RBC supernatant potassium, which may be required
prior to massive or rapid infusion of stored RBC to neonates

48. FROZEN RED CELLS
INDICATION:
• Donor red cells with rare blood groups can be stored
frozen for recipients who have developed antibodies
against frequently-occurring red cell antigens.
• -For future autologous transfusion, if blood group is
rare.
• -Associated with low risk of nonhaemolytic transfusion
reactions

49. 1
• Glycerol is gradually added to the red cells as a
cryoprotectant to a final concentration of 40%
(weight/volume).
• The cells are then frozen at -65°C or colder for up to
10 years
2 • Before transfusion, red cells are thawed and glycerol
removed by automated cell processor
3 • Can be stored in a refrigerator for up to 2
weeks after thawing.

50. Transfusion of gamma-irradiated red cells is indicated for prevention of graft vs. host
disease in susceptible individuals like:
 Immunodeficient individuals, and
 Patients receiving blood from first-degree relatives.
Gamma irradiation dose: (2500 cGy)
 Impairs the proliferative capacity of donor lymphocytes in the blood component .
Disadvantage:
 Changes in the red cell membrane that result in an increased loss of potassium
from the cell, limiting the storage time of red cell concentrates to 28 days
Irradiated Red Cells:

51. PLATELETS

52. PREPARATION OF PLATRLETS
CONCENTRATION
• Random donor platelets: Prepared from 450
ml of whole blood.
• Single donor platelets: Prepared by apheresis.

53. PLATELET RICH PLASMA
• Random donor platelets concentrate, prepared from one unit of
blood collected from one doner
Volume: 50 ml
Stored : 22-24 C(Agitation)
Shelf life : 3- 5 days
Adult dose:1 unit/10 kg(4-6 units)
Pedia dose: 10 -15 ml/ kg
Manual unit raises 5000- 10,000 platelets in an adult.

54. Steps in
Blood is collected in a triple bag system with CPDA-1
only
Whole blood is centrifused at low speed (2000rpm
x5min) at 20- 22 °C
The supernatant platelet rich plasma (PRP) is then
expressed into the attached satellite bag
The supernatant PRP is further centrifused at high
speed (4000rpm x10 min at 20-22 °C) to
concentrate the platelets
Most of the supernatant platelet poor plasma is
expressed into the another satellite bag leaving
behind 50-60 ml plasma for suspension.

55. PRECAUTIONS
• Agitation during storage helps in:
-Exchange of gases.
-Maintenance of pH.
-Reduce formation of platelets aggregates
• pH should never fall below 6 because
-Change in shape of platelets from disc to sphere.
-Pseudopod formation.
-Release of platelets granules.

57. Composition: must contain 3x10 ^11 plts in aprroximately 300ml of
plasma by AABB standards
Storage: 20-22 °C for maximum of 5 days
Dose: One apharesis platelet concentrate (3x10 ^11) i.e. equal to 6
units of single donor platelet concentrate
 This method is especially suitable if HLA-matched platelets are
required (i.e. if patient has developed refractoriness to platelet
transfusion due to the formation of alloantibodies against HLA
antigens).
 Contains fewer WBCS- less likely to cause febrile transfusion
reaction.
Plateletpheresis (Single donor
platelets )

58. 1 •A donor is connected to a blood cell separator machine
2 •whole blood is collected in an anticoagulant solution, platelets are separated and retained
3 •Remaining components are returned back to the donor.
•With this method, a large number of platelets can be obtained from a single donor
(equivalent to 6 units)

59. Indications:
• Megakaryocytic Thrombocytopenia:
• Leukemia
• Aplastic Anaemia
• Following radiotherapy or chemotherapy
• Dilutional Thrombocytopenia. Eg massive transfusion with stored blood.
• Disseminated Intravascular Coagulation.
• Functional Platelets abnormalities.
• Viral Disease associated with Thrombocytopenia; Dengue.

60. Plasma
compone
nts

61.  Plasma can be obtained either by centrifugation of a unit of
whole blood or by plasmapheresis.
 Various components can be prepared from plasma.
 Important ones being:
1. fresh frozen plasma (FFP)
2. Plasma Frozen within 24 Hours (FP24 )
3. Liquid Plasma, Thawed Plasma
4. Cryoprecipitated Antihemophilic Factor
5. Plasma, Cryoprecipitate Reduced
Plasma
components

62. FRESH FROZEN PLASMA
• Plasma removed from cells within 6-8 hours of collection is rapidly frozen
to below -30 C temperature.
• Before transfusion is necessary to thaw at 37 C .Once thawed there is
rapid deterioration of clotting factors, so used immediately after thawing.
• Dose: 12-15ml /kg
• Volume:200ml
• Stored: <-30 C
• Shelf life : 12 months

63. • Indication:
-Single or multiple clot factor deficiency.
-Severe liver diseases
-Massive transfusion

64. Steps in
preparation
1
• Blood collected in a CPDA-1
double or triple bag system.
• Bag balanced before
centrifugation and then
centrifused at 4,000 rpm for
10 min at 4°C
2
• Supernatant plasma (200-
250ml) is expressed into the
satellite bag using plasma
extractor and sealed.
3
• Labelled as FFP, date of
collection and expiry date and
then stored at -18°C or below
4
• When required for
transfusion , FFP is thawed in
plasma water at 37°C.
• Thawed FFP should be
transfused immediately
within 2hrs or stored between
1-6°C for no more than 24

66. Precaution
• During blood collection:
-Flow should be continuous
-450 ml bag should not take more than 8 mins to fill .
• During processing:
- Freezing the plasma as soon as it is separated.
-Maintaining the storage temparature.
• During thawing:
-entry port of bag should remain above the water.
-should be administered within 12 hours if kept at 2-6C.

67. CRYOPRECIPITATE
• Prepared from fresh frozen plasma . When FFP is slowly thawed at
cold temperature (1-6), an insoluble precipitate form, called
cryoprecipitate.
• Contain fibrinogen 1 , factor Vll, factorXIII and von Willebrand
factor.
• Volume: 15ml
• Stored at < -18 C
• Shelf life : up to 1 year

68. 1
• Cryoprecipitate can be prepared at
any time during 12 months from FFP.
2
• Frozen plasma is thawed in a cryo
bath at temp 4°C or in refrigerator
overnight
3
• Plasma and white precipitated protein
are obtained
• Bag is then centifused at 4000 rpm x
10 min at 4°C.
4
• Supernatant plasma is separated into
the satellite bag by plasma extractor
leaving 15ml of supernatant plasma
with cryoprecipitate for suspension.
• Refrozen (-18°C or below for 1 yr)
4
• When required for transfusion, has to
be thawed at 30-37°C and then kept
in the refrigerator at 2-6°C till
transfusion (for maximum of 6hours
Cryo
contin
ue…

69. Indication
Correction of hypofibrinogenemia (<100
mg/dl) in bleeding patients
Factor XIII , VIII replacement; not first-choice
therapy for hemophilia A.
von Willebrand disease.

72. Thank you !!!