Hematology 2

SCHOOL OF NATURAL SCIENCES
HEMAGY2 CLINICAL HEMATOLOGY 2
A Self-regulated Learning Module

SCHOOL OF NATURAL SCIENCES
For the Use of

Medical Laboratory Science Students
Prepared by:
Erlinda P. Sanchez
EP Sanchez " A Self-regulated Learning Module Page 1 of 88

TABLE OF CONTENTS Page No.
Chapter 1 Hemostasis ......................................................... 9

Unit 1.1 Primary hemostasis
Blood vessels: Functions; Disorders ................................... 11
Platelets: Functions; Disorders ...................................... 14
Laboratory Evaluation of Primary Hemostasis ........................... 18
Unit 1.2 Secondary hemostasis
Coagulation Factors .............................................. 29
Coagulation Cascade/Pathways .................................... 31
Unit 1.3 Laboratory Evaluation of Coagulation and Fibrinolysis ....................... 39
Unit 1.4 Hemostatic Disorders
Coagulation Disorders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Fibrinolytic Disorders . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Thrombotic Disorders .............................................. 47
Chapter 2 Myelodysplastic and Myeloproliferative Disorders . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Unit 2.1 Myelodysplastic disorders ............................................ 50
Unit 2.2 Leukemias; Myeloproliferative Disorders .................................. 52
Acute Leukemias ................................................ 53
Chronic Myelo- and Lympho- proliferative Disorders ...................... 57
Unit 2.3 Other Lymphocyte and Plasma Cell Neoplasms ............................ 59
Chapter 3 Red Blood Cell Disorders .............................................. 62
Unit 3.1 Microcytic, Hypochromic Anemias ...................................... 66
Unit 3.2 Macrocytic, Normochromic Anemias . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
Unit 3.3 Normocytic, Normochromic Anemias ..................................... 73
Hemolytic anemias ............................................... 74
Hemoglobinopathies .............................................. 81
Laboratory Activities
Activity No. 1 Capillary Fragility Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

Activity No. 2 Direct Platelet Count . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Activity No. 3 Platelet Estimation ............................................. 25
Activity No. 4 Bleeding Time: Ivy Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
Activity No. 5 Clot Retraction Time . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
Activity No. 6 Coagulation Time . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Activity No. 7 Prothrombin Time (PT) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Activated Partial Thromboplastin Time (APTT)
Activity No. 8 Reticulocyte Count . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Activity No. 9 Stained Blood Cell Examination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Activity No. 10 Osmotic Fragility Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
Activity No. 11 Sickle Cell Examination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
Page 2 of 88 A Self-regulated Learning Module "EP Sanchez

I. Course Code and Title: HEMAGY 2 (Coagulation and Blood Dyscrasias)
II. Course Description and Information: This is a 3-unit course with 2 units lecture and 1unit laboratory.
Hematology is a specialized branch of medical science that deals with the study of blood and blood-forming
organs, including the diagnosis, treatment, and prevention of diseases of the blood, bone marrow, hemostatic
system, and vascular system. Specifically, Hematology 2 emphasizes on coagulation and blood dyscrasias.
First part deals with hemostasis (principles and disorders), second part deals with disorders affecting blood
forming tissues (particularly the bone marrow); and the last part deals with erythrocytic disorders. Intact
foundation in Hematology will facilitate comprehension of the pathophysiology of these disorders. Thus,
students are encouraged to frequently review concepts in Hematology 1 as well as other related topics in
Biochemistry, Human Anatomy and Physiology. Correlation with topics in Clinical Microscopy and
Immunology is also necessary.
The laboratory part deals with special hematology procedures that will assist the clinicians in the
diagnosis and management of hematologic disorders. Laboratory activities aim to develop skills in
performing these hematology procedures as an application of the concepts learned in the lecture. Teacher-
made ideo and ome ideo link ill f he help pplemen den app ecia ion of he p oced e .
Other laboratory activities are given as inquiry-based (like case studies/analysis) that are intended to develop
critical thinking skills as well as to supplement the topics in the lecture.
Hematology (1 & 2) has a relative weight of 20% in the Medical Technology licensure examination with
the following table of specifications:
PRC Table of Specifications/Matrix for Licensure Examination

Hematology (20%)
1 Blood collection, anticoagulants and others (including Safety) 5%
2 Hematology tests and procedures 30%
2.1. Routine 15%
2.2. Automation 10%
2.3. Special 5%
3 Hematopoiesis, Diseases/Disorders and Reference values 40%
3.1. Hematopoiesis (in general) 6%
3.2. Erythropoiesis and RBCs 12%
3.3. Leukopoiesis and WBCs 12%
3.4. Thrombopoiesis and Platelets 10%
4 Coagulation(Principles, Procedures, Disorders and Reference values) 20%
4.1. Hemostasis - Theories/Concepts, Mechanisms 2%
4.2. Coagulation procedures/tests 8%
4.3. Coagulation factors, diseases/disorders & Reference values 10%
5 Quality assurance 5%
Total 100%
III. Requirements of the Course:
1. Regular Attendance to classes: you must attend online classes and live quizzes regularly by logging in
to our scheduled online activities. Online lectures will be done through Google meet and/or Facebook
live. Assessments shall be given through Quizziz, Pear Deck, Canvas and/or Google forms. For offline
students, your attendance will be monitored through your responses to text information and through
timely correspondence.

2. Submission of required activities: All required activities (assignments, research work, laboratory
illustrations) should be submitted on or before the given deadline. Deadlines will be posted by the
teacher in the google classroom and messenger group chat. It will also be texted to offline students.
For online students, requirements must be submitted to the google classroom forum or eache email
address (whichever will be specified during the class orientation). For offline students, requirements
must be submitted via mail or express courier (e.g. LBC, JRS) addressed to: Instructor s name, School
of Natural Sciences, University of Baguio, Baguio City.
3. Seventy percent (70%) passing score in all required activities: Quizzes, exams, assignments, research
work, laboratory illustrations.
Computation of grades:
The Course Grade is obtained by combining the lecture and laboratory grades (50%:50%) for the
subject.
Laboratory grade shall be computed as 30% enhancement activities (illustrations; research work;
case study; experiments when possible) plus 70% class standing (quizzes and exams).
The cumulative system of computing grades shall be followed. Grades computed for midterms and
finals are considered tentative. The final midterm grade is calculated by getting 1/3 of the first
grading grade plus 2/3 of the tentative midterm grade and the final grade is computed by getting 1/3
of the midterm grade plus 2/3 of the tentative final grade.
4. Study/Learning Guidelines:
a. Manage your time properly. As students of higher education (College), you are expected to be more
responsible in paying attention to course schedules, requirements, and deadlines. Schedule how
you will accomplish all the requirements in all your enrolled courses (reading the modules, reading
on research/ enhancement questions, doing assignments and laboratory illustrations) and focus
your attention when doing your tasks.
b. Read in advance. Read books. This module serves as your guideline in reading. While it will be
supplemented with online lectures, it should also be supplemented with textbooks and reference
materials. The laboratory will be supplemented by linked videos and prepared videos on test
procedures.
c. Observe proper conduct. Despite this online mode of learning, you must still maintain appropriate
behavior at all times. All standards of student conduct outlined in the University of Baguio Student
Handbook remain in full effect during this time of distance learning. Be honest in answering your
quizzes and exams. Work independently when accomplishing tasks and assignments.
d. Stay motivated. Your future depends on what you do today. Maintain a positive attitude towards
learning and enjoy a fun-learning environment despite the current circumstances.
e. Maintain a performance of high standard. Give your best in accomplishing all the assigned tasks. Do
not be complacent with just a 70% passing cut score. Remember that this is a board subject, and
the best preparation for the board/licensure examination should be during these formative years.
The board review is but supplementary to the knowledge you have already learned during your Med
Tech education.
f. Communicate properly. Promptly respond to notifications by regularly visiting our google classroom
and messenger group chat. If you have confusions or queries in any part of this module, I am here
to guide you through. Send your academic concerns using the same online platforms. For offline
students, text messages and mobile calls are welcome during scheduled hours of the day and
week. Be guided by this schedule when communicating:
Respect private hours. I do not always open my laptop/email/messenger 24/7. Send your
queries and/or concerns during regular office hours. For concerns that need immediate
attention, send through mobile text.
Be patient. Messages received between 8 AM to 8 PM will be responded to within the same day.
Messages received after 8 PM will be answered starting 8 AM the next day.
Before calling my mobile number, text first for permission for I might be giving an online lecture
or in a meeting or on private moment at that very instance.
Saturdays and Sundays are for my family and home chores. I shall respond to
queries/messages received during these days within the first office hour of Monday.

g. Show mutual support. Support one another. Let us all be responsible and supportive in making this
new learning process more effective.
h. Live lecture/Video conferencing guidelines:
h.1 Be punctual. Live lectures/Video conferences will be scheduled during the official class
period/time of this course. Log in to the platform at least 5-10 minutes before the class
period. Prepare your learning materials such as this module, pens, papers, etc. Attendance
will be checked during the lecture/video conference.
h.2 Maintain professionalism.
- Wear appropriate clothing and set your gadget in an appropriate area. You may be asked
to turn on your video/camera at any time during the lecture.
- Log in using your UB Gmail account. Unidentified names like nicknames, phone models,
etc. will not be allowed in the video conference.
- Mute your microphone as soon as you log in to the platform to avoid any excess
background noise. Unmute your microphone when instructed to do so.
- Be courteous. Do not interrupt your teacher or a classmate who is speaking. You may
pe o q e ion in he Cha a ea, o e he ai e hand fea e if a ailable, and ai
until you are allowed to speak.
- Respect privacy. Do not take a screenshot, picture, snapshot, etc. of your teacher or
fellow students, nor make any unnecessary audio or video recordings.
h.3 Remain focused and engaged. Do not be distracted by your gadget. Keep your
videoconference platform open and do not navigate other tabs or webpages unless
directed by your teacher.
i. Additional learning materials: E-books, teacher-made recorded videos and relevant video links.
Hematology E-books and teacher-made videos will be made available in the google classroom.
Relevant video links are already indicated in the laboratory activity sheets/pages. Offline students
should contact the teacher on how these materials will be shared.
j. Rubrics for grading: Research works/Essay Questions /Assignments and Laboratory Illustrations
Essay
1 2 3 4
question
Completenes Some data are left out Output contains all
Complete non- Some data are left
or answers are necessary data required
s of answer answering of a out; and there is
complete but by the activity (ex. Label).
(4pts) particular no discussion of
discussion is not The answer is complete
-per question question answer
comprehensive and comprehensive
ILLUSTRA-
1 2 3 4
TIONS
Illustration/Drawing Some illustrations/ Illustration/Drawin Output is of great
does not meet criteria Drawings are not at g is satisfactory quality.
Aesthetics
expected from the level of a student but could be Illustration/Drawing is
(4pts)
students taking taking professional improved at the level required for
professional subjects. subjects professional subjects
Complete non- Output contains all
Completeness More than 2 labels More than 1-2
illustration/drawing of necessary data
(4pts) are left out labels are left out
a particular item required by the activity.
Content is correct, no
Content More than 5 errors 1-3 errors are errors are noted (e.g.
4-5 errors are noted
(4pts) are noted noted correct color of
samples / tubes)
TOTAL

Research
0 1 2 3
Assignment
Completenes Some data are left out Output contains all
Complete non- Some data are left
or answers are necessary data required
s of answer answering of a out; and there is
complete but by the activity (ex. Label).
(4pts) particular no discussion of
discussion is not The answer is complete
-per question question answer
comprehensive and comprehensive
Answers are
Answers are Content is correct,
summarized
summarized and concise yet important
Content haphazardly; some
derived from reliable details are seen; Answers
(4pts) No Output answers are
sources but a few are derived from reliable
-per question copied verbatim, or
errors in the content of sources; there is evidence
from unreliable
the output are seen of personal input
sources
Technical
Aspects x 2 point if all Technical aspects required is met
(2pts) x 1 point if not all Technical aspects required are met
per report
TOTAL
k. Format for enhancement questions/research work/assignment:
Name: Subject/Section
Date:
Research Work No 1:
1. The question (must be highlighted) This format

applies to
Answer (Direct answer/enumeration; Discussion) EACH
Reference: Book or E-book: Title; Author; Edition; Page Number question
Must be in Internet sources: complete website address (Universal

RED font Resource Locator/URL; Date Retrieved).
Submit in WORD format
IV. Learning Competencies: After successfully completing this course, you must be able to:
1. discuss the mechanisms of hemostasis, coagulation, and fibrinolysis substantially;
2. accurately outline the coagulation pathways, the factors involved in each, and their roles in the
coagulation system;
3. correlate the importance of hematological tests in the diagnosis and prognosis of disease
4. identify the different blood disorders (RBC, WBC, Coagulation & Fibrinolysis);
5. assume responsibility in the collection and handling of blood specimens, and in the examination and
determination;
6. apply systematically the principles and procedural steps in common and special diagnostic
laboratory examination to evaluate blood disorders.
7. appreciate the correct performance of hematology procedures through video viewing.
8. develop the necessary skills such as technical, clinical, mathematical, and judgmental skills, and the
proper use of equipment and reagents; and
9. manifest responsibility, cooperation, self-reliance, honesty, critical thinking, empathy, and value for life.

V. About this Module:
This Hematology2 module is intended to guide students (both online and offline/distant learners) in
understanding Hematology2 through self-paced learning. It is a compilation of concepts from different
books/references, but is not complete in itself. It is not a textbook nor a replacement thereof. Thus, you
are encouraged to further supplement your knowledge through reading prescribed textbooks and other
reference materials. Supply the other information (with posted reading/assignment guide) through reading
of books and answer the self-assessment only after completing the topic.
TERESA N. VILLANUEVA, RMT, MACT

Dean, School od Natural Sciences
VI. First Semester; Academic Year 2020 - 2021

( August 24, 2020 December 24, 2020)
WEEK TOPIC/S ACTIVITIES
Lecture and Laboratory:
Course Orientation, 1. Read the course description, course requirements and study
Week 1
Introduction and Overview guidelines.
2. Familiarize yourself with the platforms we will be using.
Lecture:
1. Read and understand the scheduled topic
2. Participate in the online lecture
Week 2 Primary hemostasis
3. Answer the assessment quiz
Laboratory:
1. Accomplish activity no. 1
Lecture:
Secondary hemostasis 2. Participate in the online lecture
Week 3
-Coagulation factors Laboratory:
1. Accomplish activity no. 2 and 3
2. Watch the teacher-made videos
Lecture:
Secondary hemostasis
Week 4 3. Answer the assessment quiz
-Coagulation pathways Laboratory:
2. Watch the teacher-made video
Lecture:
Tests to Evaluate 2. Participate in the online lecture
Coagulation and fibrinolysis
Laboratory:
1. Accomplish activity 5 and 6
Week 6 First Grading Exam
Lecture:
Disorders affecting 2. Participate in the online lecture
Week 7
Coagulation Laboratory:
Lecture:
Disorders affecting 1. Read and understand the scheduled topic
Week 8 fibrinolysis and thrombosis 2. Answer the assessment quiz
Laboratory:
1. Analyze and answer the case studies to be given this week

Lecture:
Week 9 Myelodysplastic disorders 3. Answer the assessment
Laboratory:
1. Research study: Bone marrow and peripheral blood pictures of
the disorders.
Lecture:
Leukemias
Week10 3. Answer the assessment
Myeloproliferative disorders Laboratory:
1. Research study: Bone marrow and peripheral blood pictures
of the disorders.
Lecture:
Other lymphocyte and 2. Participate in the online lecture
Week 11 Plasma cell neoplasms
Laboratory:
1. Accomplish activity no. 8;
3. Access the shared video link
Week 12 Midterm Examination
Lecture:
Introduction to anemias 1. Read and understand the scheduled topic
Week 13 Microcytic- Hypochromic
Laboratory:
anemias
1. Accomplish activity no. 9;
Lecture:
1. Continue reading microcytic-hypochromic anemias.
Microcytic-Hypochromic
Week 14 anemias Laboratory:
3. Access the shared video link
Lecture:
Macrocytic-Normochromic 2. Participate in the online lecture
anemias
Laboratory:
1. Accomplish activity no 11
Lecture:
Normocytic-Normochromic 2. Participate in the online lecture
Week 16
anemias 3. Answer the assessment quiz
Laboratory:
1. Analyze and answer the case studies
Lecture:
Week 17 Hemoglobinopathies
Laboratory:
Collate all laboratory activities

Unit 1.1: PRIMARY HEMOSTASIS: Components, Functions, Disorders
BLOOD VESSEL (Vascular Intima)
Endothelial Cells
Page 10 of 88 A Self-regulated Learning Module " EP Sanchez
Laboratory Activity 1: CAPILLARY FRAGILITY TEST
NAME: Rating:
Class schedule: Date submitted:
This test measures the ability of small capillaries to retain blood when subjected to increased
hydrostatic pressure and anoxia. It is a non-specific evaluation to measure capillary weakness and deficiencies
in platelet number and function. Decreased capillary resistance causes the capillaries to rupture which leads to
bleeding and formation of petechiae.
Objectives: At the end of this activity, you should be able to:

properly perform the capillary fragility test (tourniquet); and
discuss the clinical significance of the test in the screening of primary hemostatic disorders.
Materials Needed: Blood pressure cuff (or tourniquet/rubber or cloth strip), timer
Procedure: Rumpel-Leede Tourniquet Test
1. Examine the forearm, hand, and fingers to make certain that no petechiae are present.
2. With a blood pressure cuff, apply 100 mmHg pressure to upper arm.
F To those who do not have a blood pressure cuff, use a tourniquet or rubber/cloth strip instead.
Apply the tourniquet not too tight, not too loose to employ just enough pressure.
3. Maintain pressure for 5 minutes.
4. Release cuff and wait for 5 10 minutes before making a final reading.
5. Examine the forearm, hands and fingers for petechiae.
Note: Disregard any petechiae within ½ inch of the blood pressure cuff (tourniquet)
because this may be due to pinching of the skin by the cuff.
6. Count the number of petechiae and roughly interpret as follows:
1+: A few petechial on the anterior part of the forearm

2+: May petechial on the anterior part of the forearm
3+: Multiple petechial over the whole arm and back of the hand
4+: Confluent petechial on the arm and back of the hand
Illustrate:
1. Petechiae, purpura, ecchymosis
2. Photo of your individual results.
Enhancement Questions:
Reading Assignment:
1. What are petechiae, ecchymosis, purpura?
2. Enumerate and describe the different vascular disorders that lead to bleeding.
3. Clinical significance of CFT
Research Question: (to be checked using the rubrics) Format or you report
1. Give the factors affecting the results of the test
2. Give and explain the contraindications of CFT.
Photo (Individual Result)
Answer to the Research questions

HEMATOLOGY 1:
Megakaryopoiesis
Functional zones of Platelets.

Platelet Aggregation Test

Laboratory Activity 2: DIRECT PLATELET COUNT
NAME: Rating:
Platelets are thin disks, 2 4 m in diame e and 5 7 fL in volume. Platelets function primarily in
hemostasis and in maintaining capillary integrity. Platelet numbers must be sufficient for them to play their
supportive role in hemostasis. When evaluating a bleeding problem that maybe traceable to platelets, the
counting of platelets is an important and logical starting point
Objectives: At the end of this activity, the students should be able to:
appreciate the proper performance of direct platelet count through viewing of video-recorded
demonstration; and
appreciate the clinical significance of platelet count in the screening of hemostatic disorders.
Materials and Reagents Needed:
Light Microscopy: Tocantins method Phase-Contrast Microscopy:

Diluting fluid: Rees & Ecker fluid Diluting fluid: 1% ammonium oxalate (freshly prepared)
RBC pipette Dilution vial
Sucking tube Phase-Contrast Microscope (or Light Microscope used
Light Microscope with subdued light
Moist Chamber: Petri dish and filter paper

Hemocytometer
Tally counter
Specimen Needed: EDTA-anticoagulated venous blood or

First few drops from a deep skin puncture with freely flowing blood
PROCEDURES:
A. LIGHT MICROSCOPY: T ca i eh d
1. Do a finger puncture or venipuncture with EDTA tube.

2. Draw first diluting fluid up to 0.5 mark of pipette then draw blood to exactly 0.5 mark and dilute to 101
mark with Rees and Ecker diluting fluid.
Note: From this point, the platelet count should be completed within 30 minutes to avoid
platelet disintegration.
3. Gently mix, discard the first 2 drops and charge into a hemocytometer.
4. Place moist chamber (Petri dish with moistened filter paper) over the charged hemocytometer and
stand for 15 minutes.
5. E amine nde high d objec i e and co n pla ele in 4 la ge co ne q a e ( W q a e ).
(Depending on the laboratory preference, platelets may be counted
either in 1/5 of a mm2 or 1 mm2 in the center of the chamber)
F Both platelets and RBCs are preserved by the Rees and Ecker
fluid. Platelets are much smaller than red cells (1/10 the size of
an RBC) and appear as round, oval or elongated particles
which are highly refractile and stain light bluish.
6. Calculate as follows:
Platelets/cu mm = number of platelets counted in 4 squares x DF x VCF

B. PHASE-CONTRAST MICROSCOPY: Brecker-Cronkite Method
1. Prepare a 1:100 dilution of the blood sample

2. Rotate on a mechanical mixer for 10 15 minutes the vial containing the suspension.
3. Load both sides of the hemocytometer in the usual manner using a separate capillary tube for each
side.
4. Cover the loaded hemocytometer with moist chamber for 15
minutes to allow settling of platelets in one optical plane.
5. Count platelets in 10 small squares of the central large square, 5
on each side of the chamber; or in 20 small squares, 10 on each
side of the chamber; or in all 50 small squares, until at least 100
platelets have been recorded.
F Only platelets are seen on the preparation. RBCs are seen

as ghosts on the background. Platelets appear round or oval
and frequently have one or more dendritic processes. Their
internal granular structure and a purple sheen allow the
platelets to be distinguished from debris which is often
refractile.
6. Calculate as follows:
Platelets/cu mm = total number of platelets counted x DF x VCF
Improved Neubauer counting chamber
Light Microscopy: Count platelets in 4 large/W squares

(squares A,B,C,D)
Phase-Contrast: Count platelets in 10 RBC/small

squares (colored squares)

Laboratory Activity 3: PLATELET ESTIMATION
NAME: Rating:
A properly prepared blood smear is utilized for platelet estimation and for the observation of any
abnormal platelet size and distribution.

appreciate the proper performance of platelet estimation through viewing of video-recorded
demonstration;
properly report the estimated platelet count; and
correlate the platelet estimate with the direct platelet counts.

Immersion oil, Microscope , Tally counter
Specimen Needed: Wright- or Giemsa-stained blood smear
PROCEDURE:
1. Select an area of the blood film in which most RBCs are

separated from one another with minimal overlapping of RBCs
and where platelets are not clumped.
2. Using the 100x oil immersion objective, count the number of
platelets in 10 consecutive fields, and calculate the average
number of platelets per field. Ba lemen pa e n of co n ing cell
3
3. To obtain the platelet estimate per µL or mm of blood, multiply the average number of platelets per field
by 20,000.
4. Report the platelet count qualitatively using the following references:
Notes:
F Accurate estimates are possible only when there are no platelet clumps, or at most, rare clumps of 2
to 3 platelets.
F A better estimate is possible using venous blood with EDTA as an anticoagulant, in which platelets
are evenly distributed and where clumping normally does not occur.
F On average, there are 8 20 platelets per field with 200 red cells.
Platelet Estimate/uL: Report as. . .
0 - 49,000 Marked decreased

50,000 - 99,000 Moderate decreased
100,000 - 149,000 Slight decreased
150,000 - 199,000 Low normal
200,000 - 400,000 Normal
401,000 - 599,000 Slight increase
600,000 - 800,000 Mod. increase
Above 800,000 Marked increased

Illustrate: (to be checked using the rubric)
1. Characteristic blood picture in Essential thrombocytosis, Gray platelet syndrome and Bernard-Soulier
syndrome.
2. Platelet satellitism
Enhancement Questions: (For laboratory activities 2 and 3)
(Reading assignment)
1. Read on functions of platelets
2. What are the platelet factors?
3. Give the compositions and function/s of each component of the Rees and Ecker fluid.
4. Describe the principle of automated platelet count
(Research questions to be checked using the rubric)

1. Give the sources of errors in platelet counting (Falsely low and falsely high counts- Manual and automated
methods)
2. Describe these principles of platelet aggregation:
a) Light-transmittance aggregometry
b) Electrical impedance
c) Optical Lumi-Aggregometer

Laboratory Activity 4: BLEEDING TIME
NAME: Rating:
Bleeding time is the time it takes for a standard wound at a standard pressure to stop bleeding. This
serves as a screening test for detecting disorders of platelet function and the ability of the small blood vessels to
control bleeding after injury.

appreciate the proper performance of Ivy bleeding time through viewing of video-recorded
demonstration; and
appreciate the clinical significance of the test in the screening of primary hemostatic disorders.
Materials and Reagents Needed: Blood pressure cuff Stopwatch

Blood lancet Filter paper strips
PROCEDURE:
1. Place a blood p e e c ff on he pa ien a m abo e he elbo .
2. Increase the pressure to 40 mmHg and hold this exact pressure for the entire procedure.
3. Cleanse lateral part of forearm with 70% ethyl alcohol. Dry.
4. Choose an area approximately three finger-width below the bend of elbow and make 2 skin punctures.
Incision must be made parallel to the elbow crease.
Note: Avoid underlying subcutaneous veins.
5. Start stopwatch as soon as blood appears from the puncture.
6. Blot the blood from each puncture with the edge of a filter paper every 30 seconds interval.
Note: Care must be taken not to touch the incision.
7. End point is when no blood comes out of the punctured area/blood does not stain the filter paper.
8. Record the bleeding time of the 2 puncture sites and report the average of the two results.
https://www.youtube.com/watch?v=bMVy6pCWhRk
Reading questions:
1. Enumerate and briefly discuss the factors affecting bleeding time.
2. Discuss the clinical significance of bleeding time
3. Read on qualitative disorders of platelets
(to be checked using the rubrics)

4. Explain the effect of aspirin on platelet function.
5. What are the precautions in performing bleeding time? Explain each.

Laboratory Activity 5: CLOT RETRACTION TIME (CRT)
NAME: Rating:
Within 1 hour after whole blood is allowed to clot in a clean glass tube at 37 OC, the clot will begin to
shrink and retract from the walls of the tube. Serum is expressed and the clot becomes denser. This retraction
process is maximal at 24 hours, by which time it occupies almost half of the original blood volume

appreciate the proper performance of clot retraction time through viewing of video-recorded
demonstration; and
appreciate the clinical significance of the test in the screening of hemostatic disorders.
Materials Needed: Test tubes (13 x 100 ) graduated cylinder Applicator sticks
Centrifuge tubes water bath
Specimen Needed: Fresh whole blood
PROCEDURE: Modified MacFarlane Serum method
1. Collect 5 ml of venous blood samples. Note the time of

collection.
2. Dispense into a graduated centrifuge tube noting exact
amount of blood used.
3. Place an applicator stick at the center of tube.
4. Incubate test tubes at 370C in a water bath for 2 hours.
5. Examine and record degree of retraction (partial,
complete, or no retraction)
6. Pull applicator stick with clotted blood and describe the
clot.
7. Serum expressed is centrifuged for 3-5 minutes at 3,500
rpm.
8. Calculate % of serum expressed as follows:
a. Volume of Serum = total volume of serum in tube volume of packed red cells
b. Then: % serum expressed = Volume of serum in ml x 100

Volume of blood used in ml.
9. Report results in %
Enhancement Questions: (to be checked using the rubrics)

1. Explain the factors that affect clot retraction
2. Discuss the clinical significance of CRT.

Thrombin Feedback Mechanism:
¾ Activation of Coagulation
It is autocatalytic or self-perpetuating;
Low level of thrombin activates V and VIII;
Activates XIII and XI
Induces platelet aggregation
¾ Inhibitor to Coagulation
Controls excessive coagulation
Increase concentration of thrombin; destroys V and VIII; activates Protein C
ª Protein C and S increase plasminogen activation
ª Promote plasmin generation (fibrinolysis

Laboratory Activity 6: WHOLE BLOOD COAGULATION TIME
NAME: Rating:
This measures the time required for blood to clot after it has been removed from the body. This is a
measure of the overall intrinsic and common pathways of coagulation.
Objectives: At the end of this activity, you must be able to:

appreciate the proper performance of whole blood clotting time through viewing of video-
recorded demonstration; and
appreciate the clinical significance of the test in the screening of secondary hemostatic
disorders.
Materials Needed: Glass slides Stopwatch

Test tubes (13 x 100mm) Water bath (370C).
Specimen required: Micro Method capillary blood

Macro Method Whole blood
PROCEDURE:
A. Micro Method slide method

1. Do a finger puncture. Start stopwatch at the time of appearance of blood
from the puncture
2. Place on a clean glass slide 3 separate drops of blood.
3. Allow to stand for 2 minutes at room temperature.
Check clot formation by drawing the blood with a needle or lancet and
observe for thread formation.
4. Record clotting time from the start to fibrin thread formation.
B. Macro Method Lee and White Method

1. Label three uniformly sized tubes 1, 2 and 3.
2. Make a clean venipuncture using 20-gauge needle and note the time at which blood enters the
syringe. Draw four (4) ml of blood.
3. Carefully dispense 1 ml to tube 3, then 1 ml to tube 2 and 1ml to tube 1. Discard the remaining
blood.
4. Incubate all tubes in a water bath at 37 0C (+ 0.5 0C )
5. At exactly 5 minutes, tilt tube number 1 to a 450 angle and observe for clotting. Repeat every 30
seconds until tube can be completely inverted without spilling contents. Note time of clotting.
6. After 30 seconds tube 1 is clotted, proceed to tube 2 and repeat the preceding procedures. Repeat
procedures with tube 3.
7. Record coagulation time as the time elapsed between the withdrawal of blood and the completion of
coagulation in tube 3.
Reading assignment:
1. Read on the different plasma coagulation factors (synonyms, functions, group, and pathway
involvement).
2. Read on the classical concept of coagulation.
To be checked using the rubric:

1. Explain the technical errors that may affect coagulation time.
2. Discuss the current concept of coagulation

FIBRIN lysis FIBRINOGEN lysis

Laboratory Activity 7: ACTIVATED PARTIAL THROMBOPLASTIN TIME and
PROTHROMBIN TIME
NAME: Rating:
Activated Partial Thromboplastin Time is a useful procedure for routine screening of coagulation disorders
in the intrinsic and common pathways.
Principle: Platelet-poor plasma contains all the coagulation factors needed for the generation of intrinsic
prothrombinase/plasma thromboplastin except Ca++ and platelet phospholipid. When Ca++ is added
with incomplete thromboplastin, intrinsic prothrombinase is generated Prothrombin is converted to
thrombin which cleaves fibrinogen into a fibrin clot.
Prothrombin Time is a useful screening procedure for the extrinsic and common pathways of coagulation.
Principle: When tissue extract or thromboplastin is added to platelet-poor plasma along with Ca++, it reacts
with factor VII, to convert factor X to Xa. This subsequently initiates the common pathway.

appreciate the proper performance of PT and APTT through viewing of video-recorded
demonstration; and
appreciate the clinical significance of these important screening tests for secondary hemostatic
disorders.

Test tubes (13 x 100) Water bath 370C 0.1 ml, and 0.2 ml pipettes
Distilled water PT & APTT Reagents 3.2% sodium citrate
Stopwatch Control plasma
Specimen Required: Citrated plasma (centrifuged at 2,500 rpm for 15 mins)
PROCEDURES: Tilt Tube Technique (Manual):
A. Prothrombin Time: Procedure Reference: QuikCoag TM ( BioMedica)

1. Pre-incubate the reconstituted PT reagent to 370C (in a water bath) for at least 10 mins. Maintain the
suspension of the reagent by magnetic stirring or mixing by inversion immediately prior to use.
2. Pipette 100 uL test plasma into a test tube and incubate at 370C for 1 minute.
Note: Do NOT incubate samples at 37OC for longer than 5 mins to avoid the loss of factors V & VIII.
3. Rapidly add 200 uL of the pre-incubated PT reagent and simultaneously start the timer.
4. Remove the test tube from the water bath; gently tilt the test tube back and forth until a gel clot forms;
stop the watch; and record the elapsed time in seconds.
5. Record the clotting time in seconds.
Expected PT ( Manual): 12 15 seconds
Note: A control plasma should be run together
with the test.
International Sensitivity Index (ISI): ____
Compute for International Normalized Ratio
(INR) using the following formula:
INR = Pa ien PT ISI
Mean Normal PT

B. Activated Partial Thromboplastic Time:
(Procedure Reference: AMS Company)
1. Pre-incubate the Calcium Chloride (0.02 M) to 370C (in a water bath) for at least 10 mins.
2. Pipette 100 uL test plasma into a test tube and incubate at 370C for 1 2 minutes.
3. Add 100 uL of the APTT reagent to the incubated plasma. (Maintain the suspension of the reagent by
magnetic stirring or mixing by inversion immediately prior to use).
4. Incubate the mixture at 37 OC for 3 minutes.
5. Rapidly add 100 uL of the pre-incubated Calcium Chloride and simultaneously start the timer. Gently
tilt back and forth until a gel clot forms.
6. Record the clotting time in seconds.
Expected APTT : 22 34 seconds
Enhancement Questions: (to be checked using the rubrics)

1. How should the plasma sample for routine coagulation tests be stored?
2. What is the purpose of computing the International Normalized Ratio?
3. What is a circulating anticoagulant? A Specific factor inhibitor?
4. Discuss the principle and importance/function of these tests? (a. Bethesda assay b. Ecarin clotting time)

1. Vitamin K deficiency will result to abnormal
A. PT (only) factor deficiencies?
B. PT and APTT A. An inherited disorder of coagulation
C. PT, APTT and Fibrinogen level B. Severe liver disease
D. PT, APTT and Thrombin time C. Dysfibrinogenemia
D. Lupus anticoagulant
2. Which of these characterize(s) Hemophilia A? 6. PT and APTT are both prolonged, and are both
A. a sex-linked disease corrected when mixed with normal plasma. The
B. with mild to severe bleeding episode most likely deficient factor is
C. with normal bleeding time and PT A. VIII
D. A and B B. V
E. A, B and C C. XI
D. IX
3. Analyze the following laboratory results: 7. If an abnormal APTT is caused by a pathological
PT = prolonged circulating anticoagulant, it will
APTT = prolonged A. be corrected with factor VIII deficient plasma
Bleeding time = Prolonged B. be corrected with factor IX deficient plasma
Platelet count = decreased C. corrected with normal plasma
Which disorder may be indicated? D. All of these
A. Factor X deficiency E. None of these
B. on Willeb and di ea e 8. Analyze the following laboratory results:
C. DIC Platelet count: 355 × 109/L
D. Factor V deficiency PT: 28 sec
4. Analyze the following laboratory results: APTT: 65 sec
PT = 23 seconds
APTT = 54 seconds
Transaminases: elevated (AST & ALT)

thrombin time = 19 seconds. These clinical presentations and laboratory
What is the most probable diagnosis? results are consistent with:
A. Factor VII deficiency A. Inherited factor VII deficiency
B. Factor II deficiency B. DIC
C. Factor X deficiency C. Liver disease
D. Hypofibrinogenemia D. on Willeb and di ea e
E. Any of these is possible E. Vitamin K deficiency
5. Which of the following is associated with multiple

WHO Classification - Main Categories of AML
1. AML with recurrent cytogenetic abnormalities
2. AML with multilineage dysplasia
3. AML, Therapy related
4. AML, Not otherwise specified (subclassified by morphology and immunophenotype).

Chapter 3
ERYTHROCYTIC DISORDERS

Laboratory Activity 8: RETICULOCYTE COUNT
NAME: Rating:
The reticulocyte is the cell stage immediately before the mature erythrocyte. It is released into the
peripheral circulation where it spends a day of maturation. The reticulocyte count, with its associated
corrections, can be used to assess bone marrow erythropoietic activity.

appreciate the proper performance of reticulocyte count through viewing of video-recorded
demonstration; and
Appreciate the clinical significance of reticulocyte count and its other associated corrections.

WBC pipettes Glass slides Brilliant Cresyl Blue or
Microscope Wright Stain New Methylene Blue
Specimen Needed: EDTA-anticoagulated blood or Capillary blood
PROCEDURE: Dry Preparation:

1. Mix equal amounts of filtered stain and EDTA-anticoagulated blood or fresh capillary blood in a small test
tube.
2. Incubate mixture at room temperature for 10 15 minutes.
3. After incubation, mix thoroughly and prepare a wedge smear.
4. Examine smear using 100x objective (Routine light microscope method). Select an area where
erythrocytes are close but not overlapping and reticulocytes appear to be well stained.
Appearance of RBCs and reticulocytes
Mature RBCs: light to medium green without granules
Reticulocytes: light green with granules that stain deep blue
5. Count the number of reticulocytes in 1000 red cells. Reticulocytes should also be counted as
erythrocytes.
6. Calculate as follows: % Reticulocyte = No. of reticulocytes x 100
1000 RBC
https://www.youtube.com/watch?v=q3oNCz_U1sY
Illustrations:
1. Routine light microscope method: One oil immersion field showing both mature erythrocytes and
reticulocytes
2. Miller disk method: One oil immersion field showing both red cell and reticulocytes with the
superimposed Miller disk. Indicate the areas for RBC and Reticulocyte counting.
F Indicate the formula for each method
Review questions/Reading Assignment:
1. Give the normal values (infants & adults) and the clinical significance of decreased and increased
reticulocyte count.
2. Describe the Miller disc. Give its formula.
3. What are the other associated corrections for reticulocytes? Give their formulas and significance.
Research Questions (to be checked using the rubrics)
1. Explain the relationship between reticulocytosis and polychromatophilia.
2. What are the indications of Reticulocytosis? Explain each
3. What are the indications of Reticulocytopenia? Explain each

Laboratory Activity 9: STAINED BLOOD CELL EXAMINATION
NAME: Rating:

correctly identify the morphological features of blood cells, both normal and abnormal.
appreciate the clinical significance of the abnormal cells.
Materials and Reagents Needed: Microscope, immersion oil
Specimen Required: Stained blood smears (CSR Control slides)
PROCEDURE:
1. Using oil immersion objective, study a thin area of the blood smear. Cells must be evenly distributed and
free of stain precipitates, not overlapping nor bunched together.
2. Examine about 15 microscopic fields and make observations of:
a. RBC hemoglobin con en , i e, hape, and p e ence of incl ion bodie .
b. WBC c opla m and n cle
c. Pla ele i e, hape and di ib ion
3. Report the degree of anisocytosis and/or poikilocytosis following the standard manner of reporting
as ( - ; +1, +2, +3 or +4 )
Illustrate (Images)
Illustrate the blood pictures in the following disorders: Describe each blood picture and label significant
findings (e.g. hallmark finding/s).
1. Severe Iron deficiency anemia
2. Hereditary spherocytosis
3. Megaloblastic anemia.
4. Myelofibrosis
5. Hemolytic anemias like G6PD deficiency, DIC or MAHA
6. Sideroblastic anemia
7. Hereditary stomatocytosis
8. Abetalipoproteinemia
Illustrate bone marrow smears from:
9. Normoblastic marrow
10. hyperplastic marrow
11. hypoplastic marrow
Format for the output:

Image/Blood picture Image/Blood picture
Description Description
Review Questions/Reading Assignment:
1. Review the descriptions and clinical significance of the different anisocytes, poikilocytes and red cell
inclusions.
2. Review the standardized manner of reporting anisocytosis, poikilocytosis and hypochromia

Laboratory Activity 10: OSMOTIC FRAGILITY TEST (OFT)
NAME: Rating:
This test measures the ability of the RBCs to take in fluid without lysing. It reflects the shape and size of
erythrocytes (specifically the surface area-to-volume ratio). Cells with decreased surface area-to-volume ratio
have a limited capacity to expand in hypotonic solutions and therefore lyse even at a less hypotonic
concentration of saline than the normal biconcave cells.

appreciate the proper performance of osmotic fragility tests through viewing of video-recorded
demonstration;
accurately determine the significant OFT results; and
appreciate the clinical significance of the test in the diagnosis of red cell disorders.

Test tubes (13 x 100) Centrifuge 0.5% NaCl
Test tube rack distilled water
Specimen Required: Heparinized or defibrinated blood
PROCEDURE: Sanford Method:
Prepare different concentrations of hypotonic saline using the following procedure:

1. Set up 12 test tubes and label no. 14 - 25.
2. Place in each tube 0.5% NaCl solution (The number of the tube corresponds to the drops of 0.5% NaCl).
3. Using the same dropper, add distilled water into each tube until a total of 25 drops (NaCl & water) are in
each tube.
4. Dispense to each tube one drop of heparinized or defibrinated blood.
Note: Deliver at the same angle directly to saline solution.
5. Mix tubes and allow to stand at room temperature or centrifuge for 1 minute at 2,500 rpm.
6. Examine each tube for initial and complete hemolysis.
Initial Hemolysis Complete Hemolysis
faint pink tinge homogenous red mixture

supernatant with button without sediment.
of intact red cells.
Determine the % concentration of NaCl solution where initial and complete hemolysis occurred.
% NaCl = test tube number x 0.02
https://www.youtube.com/watch?v=9XB9yrBQ4xg

1. Define Osmosis and Osmotic pressure.
2. What are the factors that affect OFT?
3. Discuss completely the Incubated OFT.
4. Discuss the clinical significance of OFT.
5. Give other conditions associated with spherocytosis.
Research Questions (to be checked using the rubrics)
1. Describe the spectrophotometric determination of OFT
2. Why is heparin the recommended anticoagulant for OFT?

Laboratory Activity 11: SICKLE CELL EXAMINATION
NAME: Rating:
Some hemoglobins that aggregate and have reduced solubility are capable of polymerizing and
crystallizing within the red cell causing a distortion of cell shape (sickle shape). Hb S (Sickling Hb), when fully
oxygenated is fully soluble. Polymerization and formation into tactoid crystals occur only when oxygen is
decreased at tissue level.

appreciate the proper performance of sickle cell examination through viewing of video-recorded
demonstration; and
appreciate the clinical significance of the test in the screening of some hemoglobinopathies.

rubber band glass slide petroleum jelly Test tubes
coverslip microscope Pipettes Applicator sticks/stirrer
distilled water or deionized water Lancets, needles and syringes
Freshly made 20 g/L (2% w/v) sodium metabisulphite or sodium dithionite solution
Specimen needed: Capillary blood
PROCEDURE:
A. Scriver and Waugh Method:
1. Place a rubber band around the base of the middle finger and allow staying in place for 5 minutes.
2. Make a finger puncture on the ball of the finger and place a drop of capillary blood on a slide.
3. Immediately cover with a coverslip and seal edges with petroleum jelly.
4. Incubate the preparation at room temperature. Observe for red cell sickling at hourly intervals for 2, 3
hours, or after 24 hours if desired.
5. Microscopic examination (400x): If more than 10% of the cells are sickled, the result is positive.
B. SICKLE CELL SLIDE TEST
In the absence of the HbS solubility test, the sickle cell slide test is useful in detecting sickle cells in
patients who have either sickle cell disease or sickle cell trait.
1. Weigh 0.1g of sodium metabisulphite and transfer to a test tube capable of holding 15 ml of water.
2. Add 5 ml of distilled or deionized water, stopper, and mix until the chemical is fully dissolved. The
chemical can only be used within the day it was suspended (within 8 hrs)
3. Deli e one d op of pa ien capilla blood o ell mi eno blood on a lide and add an eq al
volume of freshly made reagent, mix and cover with a cover glass. Exclude
any air bubbles.
4. Place the slide in a plastic box or Petri dish with damp piece of blotting
paper or tissue at the bottom to prevent drying of the preparation (moist
chamber). Leave at room temperature.
5. After 10-20 minutes, examine the preparation microscopically for sickle
cells. Focus the cells first with the 10x objective and examine for sickling
using the 40xobjective. Examine several fields. Sickling usually takes place
in one part of the preparation than the other.
6. Sickle cells usually appear crescent shape with pointed ends or holly leaf
hape. Repo a ickle cell e po i i e hen c e cen hape cell a e
een, o ickle cell e nega i e hen cell appea o nded o o al
shape.

C. Tube Solubility Test
The principle of solubility method was based on turbidity created when Hb

S is incubated with sodium dithionate.
1. Add 20 uL of blood with 2 ml of the reagent prepared in procedure B (2%

w/v sodium metabisulphite or sodium dithionite)
2. Mix and stand at room temperature for 5 minutes.
3. Examine the tubes using a white board with black lines as the background
in ambient light settings.
4. Interpret results as positive if the black lines are not visible.
1. How do red cells undergo sickling?
2. Pathophysiology of sickle cell anemia.
3. Difference between sickle cell trait and sickle cell anemia.
4. Descriptions of the hemoglobin variants Hb C and Hb SC
Research Questions (to be checked using the rubrics):
1. What are the sickling hemoglobins?
2. Explain the influence of HbS gene with Plasmodium infection.
3. Describe the hemoglobin variant HbE.

Self-Assessment: Instruction: Answer completely the following
Note: Graded quizzes will be given via the indicated online platforms during regular meetings. For offline
students, essay questions will be sent via available correspondence.
1.
End of Hematology 2
End of Hematology 2

EVALUATION OF THIS COURSE:
Dear students:
Please evaluate this course (HEMAGY1) honestly and objectively. Rest assured that your responses
will be taken positively and reflectively for the improvement of this course.
1. What lesson or activity did I enjoy most? Why?
2. What is the most important lesson which I can apply in my daily life?
3. What are the new insights/discoveries that I learned?
4. What topic/s do I find least important?
5. What possible topics should have been included?
REFERENCES:
1. Brown, B. (1993). Hematology: Principles and Procedures (6th ed.) Philadelphia: Lea & Febiger
2. Carr, J., & Rodak, B. (2016). Clinical Hematology Atlas (5th ed.). Philadelphia: W.B. Saunders.
3. Ciesla B., (2012), Hematology in Practice (2nd ed.). Philadelphia: F.A. Davis Co.
4. Mcpherson, R. A. & Pincus, M. R., (2011). Hen Clinical Diagn i and Managemen b Lab a
Methods. (22nd ed.) Philadelphia: Elsevier Inc..
5. Steininger, C., (1992). Clinical Hematology Principles Procedures Correlations. Philadelphia:

J. B. Lippincott Co.
6. Turgeon, Mary. (2012). Clinical Hematology, Theory and Procedures. (5th ed.) Lippincott Williams &
Wilkins.


Hematology 2

Uploaded by

Copyright:

Available Formats

Hematology 2

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Hematology 2

Uploaded by

Copyright:

Available Formats

SCHOOL OF NATURAL SCIENCES

HEMAGY2 CLINICAL HEMATOLOGY 2

A Self-regulated Learning Module

For the Use of

EP Sanchez " A Self-regulated Learning Module Page 1 of 88

Chapter 1 Hemostasis ......................................................... 9

Activity No. 1 Capillary Fragility Test . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

Page 2 of 88 A Self-regulated Learning Module "EP Sanchez

PRC Table of Specifications/Matrix for Licensure Examination

III. Requirements of the Course:

EP Sanchez " A Self-regulated Learning Module Page 3 of 88

Page 4 of 88 A Self-regulated Learning Module "EP Sanchez

EP Sanchez " A Self-regulated Learning Module Page 5 of 88

k. Format for enhancement questions/research work/assignment:

1. The question (must be highlighted) This format

Must be in Internet sources: complete website address (Universal

Submit in WORD format

Page 6 of 88 A Self-regulated Learning Module "EP Sanchez

TERESA N. VILLANUEVA, RMT, MACT

VI. First Semester; Academic Year 2020 - 2021

EP Sanchez " A Self-regulated Learning Module Page 7 of 88

Page 8 of 88 A Self-regulated Learning Module "EP Sanchez

BLOOD VESSEL (Vascular Intima)

Class schedule: Date submitted:

Objectives: At the end of this activity, you should be able to:

Procedure: Rumpel-Leede Tourniquet Test

6. Count the number of petechiae and roughly interpret as follows:

1+: A few petechial on the anterior part of the forearm

Answer to the Research questions

EP Sanchez " A Self-regulated Learning Module Page 13 of 88

Page 14 of 88 A Self-regulated Learning Module " EP Sanchez

Page 20 of 88 A Self-regulated Learning Module " EP Sanchez

Class schedule: Date submitted:

Materials and Reagents Needed:

Light Microscopy: Tocantins method Phase-Contrast Microscopy:

Moist Chamber: Petri dish and filter paper

Specimen Needed: EDTA-anticoagulated venous blood or

1. Do a finger puncture or venipuncture with EDTA tube.

Platelets/cu mm = number of platelets counted in 4 squares x DF x VCF

EP Sanchez " A Self-regulated Learning Module Page 23 of 88

1. Prepare a 1:100 dilution of the blood sample

F Only platelets are seen on the preparation. RBCs are seen

Platelets/cu mm = total number of platelets counted x DF x VCF

Improved Neubauer counting chamber

 Light Microscopy: Count platelets in 4 large/W squares

 Phase-Contrast: Count platelets in 10 RBC/small

Page 24 of 88 A Self-regulated Learning Module " EP Sanchez

Class schedule: Date submitted:

Objectives: At the end of this activity, you should be able to:

Materials and Reagents Needed:

Specimen Needed: Wright- or Giemsa-stained blood smear

1. Select an area of the blood film in which most RBCs are

Platelet Estimate/uL: Report as. . .

0 - 49,000 Marked decreased

EP Sanchez " A Self-regulated Learning Module Page 25 of 88

Enhancement Questions: (For laboratory activities 2 and 3)

(Research questions to be checked using the rubric)

Page 26 of 88 A Self-regulated Learning Module " EP Sanchez

Light Microscopy: Count platelets in 4 large/W squares

Phase-Contrast: Count platelets in 10 RBC/small

Expected APTT : 22 34 seconds