This document provides information on determining total leukocyte count, including causes that can lead to increases or decreases in count. The total leukocyte count is determined using a hemocytometer to count white blood cells in a diluted blood sample under a microscope. Differential white blood cell counts are also performed to determine the percentages of neutrophils, lymphocytes, monocytes, eosinophils, and basophils by examining a stained blood smear under high power magnification. Precise procedures are described for making blood smears, staining, and counting cells to obtain reliable total and differential white blood cell counts.
CONCENTRATIONS TECHNIQUES IN PARASITOLOGY PRESENTATION.pptxShreyayadav91
INTRODUCTION
Concentration procedure separate parasites from fecal debris and increase the chances of detecting parasitic organisms when these are in small numbers.
If number of organisms in stool specimen is low, examination of a direct wet mount may not detect parasites.
Thus, whenever possible, the stool should be concentrated.
Advantages
Maximizes the numbers of organisms detected which may be too scanty to be seen by direct microscopy alone. Worm eggs, larvae, and protozoan cysts may be recovered.
Disadvantages
Destroys trophozoite stages. Most concentration methods destroy trophozoites stages.
Concentration techniques can be classified as the floatation or sedimentation methods.
Floatation technique
Here solutions with higher specific gravity than the organisms to be floated so that the organisms rise to the top and debris sink to the bottom.
Principle
This technique involves suspending the specimen in a medium of greater density than that of the helminthic eggs and protozoan cysts.
Eggs and cysts float to the top and are collected by placing a glass slides on the surface of the meniscus at the top of the tube.
Floatation Methods includes:
Saturated salt solution technique
Zinc sulfate centrifugal floatation
Sugar floatation technique
Saturated salt solution technique
Procedure:
About half tea spoon (about 4 gm) of fresh stool or preserved stool in a flat bottomed container with 20 ml capacity.
Now, few drops of saturated salt solution (specific gravity 1.20) is added and stirred to make a fine emulsion.
More salt solution is added with stirring throughout to fill the container up to the brim, until a convex meniscus is formed.
A glass slide (3”*2”) is carefully laid on the top of the container so that the center is in contact with the fluid.
This preparation is allowed to stand for 20 minutes after which the glass slide is quickly lifted and examined under microscope after putting coverslip.
Zinc sulfate centrifugal floatation
Procedure
Make a fine suspension of about 1 g of feces in 10 m L of water and strain through gauze to remove coarse particles.
Collect the liquid in a small test tube and centrifuge for 1 minute at 2,500 revolutions per minute. Pour off the supernatant, add water, resuspend, and centrifuge in the same manner, repeating the process, till the supernatant is clear.
Pour off the clear supernatant, add a small quantity of zinc sulfate solution (specific gravity 1.18- 1.2) and resuspend the sediment well.
Add zinc sulfate solution to a little below the brim and centrifuge at 2,500 revolution per minute for 1 minute.
Take samples care fully from the surface, using a wire loop, transfer to slide and examine under the microscope. A drop of dilute iodine helps to bring out the protozoan cysts in a better way.
This technique is useful for protozoan cysts and eggs of nematodes and small tapeworms, but it does not detect unfertilized roundworm eggs, nematode larvae, and eggs of most trematodes and large tapeworms.
CONCENTRATIONS TECHNIQUES IN PARASITOLOGY PRESENTATION.pptxShreyayadav91
INTRODUCTION
Concentration procedure separate parasites from fecal debris and increase the chances of detecting parasitic organisms when these are in small numbers.
If number of organisms in stool specimen is low, examination of a direct wet mount may not detect parasites.
Thus, whenever possible, the stool should be concentrated.
Advantages
Maximizes the numbers of organisms detected which may be too scanty to be seen by direct microscopy alone. Worm eggs, larvae, and protozoan cysts may be recovered.
Disadvantages
Destroys trophozoite stages. Most concentration methods destroy trophozoites stages.
Concentration techniques can be classified as the floatation or sedimentation methods.
Floatation technique
Here solutions with higher specific gravity than the organisms to be floated so that the organisms rise to the top and debris sink to the bottom.
Principle
This technique involves suspending the specimen in a medium of greater density than that of the helminthic eggs and protozoan cysts.
Eggs and cysts float to the top and are collected by placing a glass slides on the surface of the meniscus at the top of the tube.
Floatation Methods includes:
Saturated salt solution technique
Zinc sulfate centrifugal floatation
Sugar floatation technique
Saturated salt solution technique
Procedure:
About half tea spoon (about 4 gm) of fresh stool or preserved stool in a flat bottomed container with 20 ml capacity.
Now, few drops of saturated salt solution (specific gravity 1.20) is added and stirred to make a fine emulsion.
More salt solution is added with stirring throughout to fill the container up to the brim, until a convex meniscus is formed.
A glass slide (3”*2”) is carefully laid on the top of the container so that the center is in contact with the fluid.
This preparation is allowed to stand for 20 minutes after which the glass slide is quickly lifted and examined under microscope after putting coverslip.
Zinc sulfate centrifugal floatation
Procedure
Make a fine suspension of about 1 g of feces in 10 m L of water and strain through gauze to remove coarse particles.
Collect the liquid in a small test tube and centrifuge for 1 minute at 2,500 revolutions per minute. Pour off the supernatant, add water, resuspend, and centrifuge in the same manner, repeating the process, till the supernatant is clear.
Pour off the clear supernatant, add a small quantity of zinc sulfate solution (specific gravity 1.18- 1.2) and resuspend the sediment well.
Add zinc sulfate solution to a little below the brim and centrifuge at 2,500 revolution per minute for 1 minute.
Take samples care fully from the surface, using a wire loop, transfer to slide and examine under the microscope. A drop of dilute iodine helps to bring out the protozoan cysts in a better way.
This technique is useful for protozoan cysts and eggs of nematodes and small tapeworms, but it does not detect unfertilized roundworm eggs, nematode larvae, and eggs of most trematodes and large tapeworms.
Physicians working in the field of hematology are called hematologists. Initially, hematologists complete a four-year medical degree and this is followed by three or four years in an internship or residency program. Thereafter, they spend two or three more years learning how to diagnose and treat blood disorders.
Incineration is the method of choice for treating large volumes of infectious waste, animal carcasses, and contaminated bedding materials. Because incinerators usually are located some distance from the laboratory, additional precautions for handling and packaging of infectious waste are necessary.
Types of Biomedical Waste Disposal
Autoclaving. The process of autoclaving involves steam sterilization. ...
Incineration. The major benefits of incineration are that it is quick, easy, and simple. ...
Chemicals. When it comes to liquid waste, a common biomedical waste disposal method can be chemical disinfection. ...
Microwaving.
Prokaryotes are always unicellular, while eukaryotes are often multi-celled organisms. Additionally, eukaryotic cells are more than 100 to 10,000 times larger than prokaryotic cells and are much more complex. The DNA in eukaryotes is stored within the nucleus, while DNA is stored in the cytoplasm of prokaryotes
Difference between prokaryotic and eukaryotic cell.pptxAmjad Afridi
Eukaryotic cells have several other membrane-bound organelles not found in prokaryotic cells.
These include the mitochondria (convert food energy into adenosine triphosphate, or ATP, to power biochemical reactions); rough and smooth endoplasmic reticulum ,golgi complex and in the case of plant cells, chloroplasts
All of these organelles are located in the eukaryotic cell's cytoplasm.
Mycology is the branch of biology concerned with the study of fungi.
The word 'myco' is derived from the Greek word mýkēs meaning “mushroom, fungus”.
Heinrich Anton de Bary is the father of Mycology.
Fungi are eukaryotic organisms that include such as yeasts, moulds and mushrooms. These organisms are classified under kingdom fungi.
Fungi are diverse and widespread.
Fungi metabolism consists on a series of reactions (biochemical reactions) constantly occurring inside the cells to keep it alive and active and in the results biosynthesis of a huge number of compounds.
These compounds area usually divided into primary and secondary metabolites.
Primary metabolism is common to several species and usually produces compounds with the function of assuring fungi growth and development.
Primary metabolites are involved in the growth, development, and reproduction of organisms.
The primary metabolites consist of vitamins, amino acids, nucleosides and organic acids
Staphylococcus aureus is a bacterium that causes staphylococcal food poisoning, a form of gastroenteritis with rapid onset of symptoms. S. aureus is commonly found in the environment (soil, water and air) and is also found in the nose and on the skin of humans.
Communicable diseases are illnesses that spread from one person to another or from an animal to a person, or from a surface or a food. Diseases can be transmitted during air travel through: direct contact with a sick person. respiratory droplet spread from a sick person sneezing or coughing.
Host-Parasite relationship is the extreme case of animal association, in which both partners influence each others life by affecting each others metabolism and behaviour using different adaptive mechanisms in order to ensure their survival.
Bacteria have their own enzymes for
1. Cell wall formation
2. Protein synthesis
3. DNA replication
4. RNA synthesis
5. Synthesis of essential metabolites
Infections spread from animals to human are called zoonotic infections.
The term zoonos is’ Derived from the Greek
ZOON (animals) and NOSES (diseases)
Pathogens shared with wild or domestic animals cause more than 60% of infectious diseases in man.
Ozone (O3) is a molecule made up of three atoms of oxygen (O), and very reactive gas.
Bluish gas that harmful to breathe.
Is mostly found in the stratosphere, where it protects us from the Sun’s harmful ultraviolet (UV) radiation.
Although it represents only a tiny fraction of the atmosphere, ozone is essential for life on Earth.
Ozone in the stratosphere— a layer of the atmosphere between 15 and 50 kilometers (10 and 31 miles) above us—acts as a shield to protect Earth’s surface from the sun’s harmful ultraviolet radiation.
H: Infects only Human beings
I: Immunodeficiency Virus weakness the Immune system and increases the risk of infections
V: Virus that attacks the body and finally kills the body’s immune system
Tuberculosis is a communicable chronic granulomatous disease caused by Mycobacterium tuberculosis , where the center of the granuloma is Caseous necrosis
It usually involves the lungs but may affect any organ or tissue in the body
Airborne spread of droplet nuclei
Lung Cancer: Artificial Intelligence, Synergetics, Complex System Analysis, S...Oleg Kshivets
RESULTS: Overall life span (LS) was 2252.1±1742.5 days and cumulative 5-year survival (5YS) reached 73.2%, 10 years – 64.8%, 20 years – 42.5%. 513 LCP lived more than 5 years (LS=3124.6±1525.6 days), 148 LCP – more than 10 years (LS=5054.4±1504.1 days).199 LCP died because of LC (LS=562.7±374.5 days). 5YS of LCP after bi/lobectomies was significantly superior in comparison with LCP after pneumonectomies (78.1% vs.63.7%, P=0.00001 by log-rank test). AT significantly improved 5YS (66.3% vs. 34.8%) (P=0.00000 by log-rank test) only for LCP with N1-2. Cox modeling displayed that 5YS of LCP significantly depended on: phase transition (PT) early-invasive LC in terms of synergetics, PT N0—N12, cell ratio factors (ratio between cancer cells- CC and blood cells subpopulations), G1-3, histology, glucose, AT, blood cell circuit, prothrombin index, heparin tolerance, recalcification time (P=0.000-0.038). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT early-invasive LC (rank=1), PT N0—N12 (rank=2), thrombocytes/CC (3), erythrocytes/CC (4), eosinophils/CC (5), healthy cells/CC (6), lymphocytes/CC (7), segmented neutrophils/CC (8), stick neutrophils/CC (9), monocytes/CC (10); leucocytes/CC (11). Correct prediction of 5YS was 100% by neural networks computing (area under ROC curve=1.0; error=0.0).
CONCLUSIONS: 5YS of LCP after radical procedures significantly depended on: 1) PT early-invasive cancer; 2) PT N0--N12; 3) cell ratio factors; 4) blood cell circuit; 5) biochemical factors; 6) hemostasis system; 7) AT; 8) LC characteristics; 9) LC cell dynamics; 10) surgery type: lobectomy/pneumonectomy; 11) anthropometric data. Optimal diagnosis and treatment strategies for LC are: 1) screening and early detection of LC; 2) availability of experienced thoracic surgeons because of complexity of radical procedures; 3) aggressive en block surgery and adequate lymph node dissection for completeness; 4) precise prediction; 5) adjuvant chemoimmunoradiotherapy for LCP with unfavorable prognosis.
Tom Selleck Health: A Comprehensive Look at the Iconic Actor’s Wellness Journeygreendigital
Tom Selleck, an enduring figure in Hollywood. has captivated audiences for decades with his rugged charm, iconic moustache. and memorable roles in television and film. From his breakout role as Thomas Magnum in Magnum P.I. to his current portrayal of Frank Reagan in Blue Bloods. Selleck's career has spanned over 50 years. But beyond his professional achievements. fans have often been curious about Tom Selleck Health. especially as he has aged in the public eye.
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Introduction
Many have been interested in Tom Selleck health. not only because of his enduring presence on screen but also because of the challenges. and lifestyle choices he has faced and made over the years. This article delves into the various aspects of Tom Selleck health. exploring his fitness regimen, diet, mental health. and the challenges he has encountered as he ages. We'll look at how he maintains his well-being. the health issues he has faced, and his approach to ageing .
Early Life and Career
Childhood and Athletic Beginnings
Tom Selleck was born on January 29, 1945, in Detroit, Michigan, and grew up in Sherman Oaks, California. From an early age, he was involved in sports, particularly basketball. which played a significant role in his physical development. His athletic pursuits continued into college. where he attended the University of Southern California (USC) on a basketball scholarship. This early involvement in sports laid a strong foundation for his physical health and disciplined lifestyle.
Transition to Acting
Selleck's transition from an athlete to an actor came with its physical demands. His first significant role in "Magnum P.I." required him to perform various stunts and maintain a fit appearance. This role, which he played from 1980 to 1988. necessitated a rigorous fitness routine to meet the show's demands. setting the stage for his long-term commitment to health and wellness.
Fitness Regimen
Workout Routine
Tom Selleck health and fitness regimen has evolved. adapting to his changing roles and age. During his "Magnum, P.I." days. Selleck's workouts were intense and focused on building and maintaining muscle mass. His routine included weightlifting, cardiovascular exercises. and specific training for the stunts he performed on the show.
Selleck adjusted his fitness routine as he aged to suit his body's needs. Today, his workouts focus on maintaining flexibility, strength, and cardiovascular health. He incorporates low-impact exercises such as swimming, walking, and light weightlifting. This balanced approach helps him stay fit without putting undue strain on his joints and muscles.
Importance of Flexibility and Mobility
In recent years, Selleck has emphasized the importance of flexibility and mobility in his fitness regimen. Understanding the natural decline in muscle mass and joint flexibility with age. he includes stretching and yoga in his routine. These practices help prevent injuries, improve posture, and maintain mobilit
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
- Video recording of this lecture in English language: https://youtu.be/lK81BzxMqdo
- Video recording of this lecture in Arabic language: https://youtu.be/Ve4P0COk9OI
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
Ethanol (CH3CH2OH), or beverage alcohol, is a two-carbon alcohol
that is rapidly distributed in the body and brain. Ethanol alters many
neurochemical systems and has rewarding and addictive properties. It
is the oldest recreational drug and likely contributes to more morbidity,
mortality, and public health costs than all illicit drugs combined. The
5th edition of the Diagnostic and Statistical Manual of Mental Disorders
(DSM-5) integrates alcohol abuse and alcohol dependence into a single
disorder called alcohol use disorder (AUD), with mild, moderate,
and severe subclassifications (American Psychiatric Association, 2013).
In the DSM-5, all types of substance abuse and dependence have been
combined into a single substance use disorder (SUD) on a continuum
from mild to severe. A diagnosis of AUD requires that at least two of
the 11 DSM-5 behaviors be present within a 12-month period (mild
AUD: 2–3 criteria; moderate AUD: 4–5 criteria; severe AUD: 6–11 criteria).
The four main behavioral effects of AUD are impaired control over
drinking, negative social consequences, risky use, and altered physiological
effects (tolerance, withdrawal). This chapter presents an overview
of the prevalence and harmful consequences of AUD in the U.S.,
the systemic nature of the disease, neurocircuitry and stages of AUD,
comorbidities, fetal alcohol spectrum disorders, genetic risk factors, and
pharmacotherapies for AUD.
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
New Directions in Targeted Therapeutic Approaches for Older Adults With Mantl...i3 Health
i3 Health is pleased to make the speaker slides from this activity available for use as a non-accredited self-study or teaching resource.
This slide deck presented by Dr. Kami Maddocks, Professor-Clinical in the Division of Hematology and
Associate Division Director for Ambulatory Operations
The Ohio State University Comprehensive Cancer Center, will provide insight into new directions in targeted therapeutic approaches for older adults with mantle cell lymphoma.
STATEMENT OF NEED
Mantle cell lymphoma (MCL) is a rare, aggressive B-cell non-Hodgkin lymphoma (NHL) accounting for 5% to 7% of all lymphomas. Its prognosis ranges from indolent disease that does not require treatment for years to very aggressive disease, which is associated with poor survival (Silkenstedt et al, 2021). Typically, MCL is diagnosed at advanced stage and in older patients who cannot tolerate intensive therapy (NCCN, 2022). Although recent advances have slightly increased remission rates, recurrence and relapse remain very common, leading to a median overall survival between 3 and 6 years (LLS, 2021). Though there are several effective options, progress is still needed towards establishing an accepted frontline approach for MCL (Castellino et al, 2022). Treatment selection and management of MCL are complicated by the heterogeneity of prognosis, advanced age and comorbidities of patients, and lack of an established standard approach for treatment, making it vital that clinicians be familiar with the latest research and advances in this area. In this activity chaired by Michael Wang, MD, Professor in the Department of Lymphoma & Myeloma at MD Anderson Cancer Center, expert faculty will discuss prognostic factors informing treatment, the promising results of recent trials in new therapeutic approaches, and the implications of treatment resistance in therapeutic selection for MCL.
Target Audience
Hematology/oncology fellows, attending faculty, and other health care professionals involved in the treatment of patients with mantle cell lymphoma (MCL).
Learning Objectives
1.) Identify clinical and biological prognostic factors that can guide treatment decision making for older adults with MCL
2.) Evaluate emerging data on targeted therapeutic approaches for treatment-naive and relapsed/refractory MCL and their applicability to older adults
3.) Assess mechanisms of resistance to targeted therapies for MCL and their implications for treatment selection
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
1. how to Determining the Total Leukocyte Count
Total Leukocyte Count by Hemocytometer
The degree of rise in leukocytes depend on the type and severity of the infection and the
response of the body.
Clinical Significance
Increase in total leukocyte count of more than 10,000/cu mm (μl) is known as leukocytosis and
decrease of less than 4 000 cu mm (μl) as leukopenia.
Causes of leukocytosis
Pathological
It is common for a transient period in infections. The degree of rise in leukocytes depend on the
type and severity of the infection and the response of the body. The infection may be 1) bacterial
2) viral 3) protozoal (malaria) or 4) parasitic (filaria, hookworm infection). Leukocytosis is also
observed in severe hemorrhage and in Leukemia.
Physiological
1. Age: At birth the total leukocyte count is about 18,000/cu mm (μl). It drops gradually to
adult level.
2. Pregnancy: At ‘full term’ the total count tends to be about 12,000 to 15,000/cu mm (μl).
It rises soon after delivery and then gradually returns to normal.
3. High temperature.
4. Severe pain.
5. Muscular exercise.
Causes of Leukopenia
Certain viral and bacterial infections (typhoid) lead to leukopenia rather than leukocytosis.
1. Infections
a. Bacterial (typhoid. paratyphoid, tuberculosis, etc.)
b. Viral (hepatitis, influenza, measles, etc.)
c. Protozoal (malaria)
2. Some cases of Leukemia.
3. Primary bone marrow depression (Aplastic anemia).
4. Secondary bone marrow depression (due to drugs, radiation, etc.)
2. 5. Anemia (Iron deficiency Megaloblastic etc)
Normal values
• Adults : 4,000-10,000/cu mm (μl)
• At birth : 10,000-25,000/cu mm (μl)
• 1 to 3 years : 6,000-18,000/cu mm (μl)
• 4 to 7 years : 6,000-15,000/cu mm (μl)
• 8 to 12 years: 4,500-13,500/cu mm (μl)
Specimens
1. Double oxalated or EDTA blood
2. Capillary blood (specimen need not be a fasting sample).
Requirements
1. Microscope
2. Improved Neubauer Chamber
3. WBC pipette
4. WBC diluting fluid: It is prepared as follows:
a) Glacial acetic acid: 2.0 ml
b) 1 % (w/v) gentian violet: 1.0 ml
c) Distilled water: 97 ml
This solution is stable at room temperature (25°C ± 5°C). A pinch of thymol may be added as
preservative.
Principle
The glacial acetic acid lyses the red cells while the gentian violet slightly stains the
nuclei of the leukocytes. The blood specimen is diluted 1:20 in a WBC pipette with
the diluting fluid and the cells are counted under low power of the micro scope by
using a counting chamber. The number of cells in undiluted blood are reported per
cu mm (μl) of whole blood.
Procedure
1. Draw blood up to 0.5 mark of a WBC pipette.
2. Carefully, wipe excess blood outside the pipette by using cotton. Draw diluting fluid up
to 11 mark.
3. 3. Mix the contents in the pipette and after five minutes by discarding few drops, fill the
counting chamber and allow the cells to settle for two to three minutes.
a. Since Bulb pipettes are not recommended following procedure is performed
b. Make a 1:20 dilution of blood by adding 20 μl of blood to (1 38 ml of diluting
fluid in a glass tube (10 * 75 mm) Cork the tube tightly and mix the suspension by
rotating in a cell-suspension mixer for at least 1 minute. Fill the Neubauer
counting chamber by means of a Pasteur pipette or glass capillary.
4. Focus on one of the ‘W’ marked areas (each having 16 small squares) by
5. turning objective to low power. (10 X).
6. Count cells in all four W marked corner.
7. Calculations
Number of white cells/cu mm (μl) of whole blood = (number of white cells
counted * dilution) / (area counted * depth of fluid)
Where: Dilution = 20
Area counted 4 * 1 sq.mm = 4 sq.mm
Depth of fluid = 0.1 mm (constant)
Hence number of white cells per cu mm (μl) of whole blood = (No. of cells counted * 20) / (4 *
0.1)
= No of cells counted * 50
· No. of leukocyte per liter of blood = No.of cells per cu mm(ml) * 106 or use the following
formula
WBC count / liter = [No. of cells counted (per 1mm2 area) / volume counted (μl)] * dilution *
106
Note: The precautions taken are exactly the same as for REC counting technique. The sources of
error are also same as for RBC counting technique. However; in the case of WBC counting extra
care is taken during the preparation and storage of WBC diluting fluid. It should be perfectly free
from dust particles and yeast cells, otherwise falsely high counts are obtained due to the presence
of yeast cells and dust particles.
Error of the total white cell count
1) The inherent distribution error = λ1/2, here λ = total number of cells in each area.
2) The error as high as 20% may make difference between 5.0 and 6.0 x 109 cells
4. per liter, which is of little practical significance.
3) The error can be reduced by counting more cells. If 400 cells are counted the
error is reduced to 5%.
4) Error may also be caused due to dirt clumped RBC debris or due to clumping of
leukocytes.
Study of blood smear for differential leukocyte count and cell morphology
Introduction
Differential count is the percent distribution of various white cells in the peripheral blood. It is
determined from a blood smear stained with a polychromatic stain and after examination of the
stained smear by using oil immersion objective (total magnification 1000 X). The number of
each type of white cell is then expressed as a percentage of the total number of cells. The stained
blood smear also helps to study abnormal morphology of leukocytes and red cells. Study of
blood smear helps in the diagnosis of various anemias, leukemias and detection of blood
parasites. Three major steps involved in differential count are –
a) preparation of blood smear
b) staining of the blood smear and
c) microscopic examination of the stained smear.
The different types of leukocytes seen in a normal peripheral blood film may be
divided into three broad groups —
1. Granulocytes
2. Monocytes and
3. Lymphocytes.
1) Granulocytes: There are three types of granulocytes which derive their names
from the staining reaction of the granules present in the cytoplasm. These cells are
a. Neutrophils or polymorphonuclears
b. Eosinophils and
c. Basophils.
(a) Neutrophils (Polymorphonuclears): The average diameter is 10 to 12 μm. The
nucleus is usually divided into 1-4 lobes. The cell derives its name from the color
of the granules. The number of lobes give an indication of the age of the cell.
(b) Eosinophils: The diameter is about the same as the neutrophil. The cytoplasm
5. contains large, oval or round, red-orange (eosinophilic) granules. The nucleus
shows fewer lobes, on an average only two.
(c) Basophils or mast cells: The diameter is 8 to 10 μm. The nucleus is not easily
seen due to the presence of large, round, deep blue or black granules.
2) Monocyte: The diameter is about 16 to 22μm. The nucleus is kidney shaped or horse shoe
shaped. Sometimes it may be round or oval. It stains pale violet and has fine chromatic
arrangement. The cytoplasm is plentiful and stains pale greyish blue and contains a number of
very fine pinkish-blue granules.
3) Lymphocyte: Two forms observed are —
a) Large lymphocyte and
b) Small lymphocyte.
(a) Large lymphocytes: It is about 12 to 15μm in diameter. This has abundant clearpale blue
cytoplasm and a large round or slightly in dented nucleus with dense chromatin.
(b) Small lymphocyte: These are small round cells about 10 to 12μm in diameter.
It has very little blue cytoplasm and often little more than just a rim around the nucleus. The
nucleus is dark, round and sometimes indented.
Differential leukocyte count technique
Differential white cell counts are usually performed by visual examination of blood films which
are pre pared on glass slides by “spread” technique. Following are the requirements for reliable
results –
1. The film should not be very thin and the tail of the film should be smooth. To achieve this, the
film should be made with a rapid movement using a smooth glass (slide) spreader.
2. In a good blood film, there is some overlap of the red cells, diminishing to separation near the
tail.
3. If rough-edged spreader is used or if the film is too thin, many of the white cells (sometimes
more than 50%), accumulate at the edges and in the tail. The distribution of white cells appears
irregular. Polymorphonuclear neutrophils and monocytes predominate at the margins and the tail,
and lymphocytes appear mainly at the middle of the film. The separation of cells depends upon
differences in stickiness, size and specific gravity among the different classes of cells.
6. 4. Some parasites (Borrelia, trypanosomes and microfilariae) can be detected in a fresh wet blood
film by their motility, but for species identification a permanent preparation is necessary. A thick
film is mainly employed for detecting parasites in the blood.
Reporting the differential white cell count
1) The differential count should be expressed as the percentage of each type of cell.
It also should be related to the total leukocyte count and then the results should be reported in
absolute numbers (x 109/l)
2) Nucleated red blood cells (NRBC) are reported as NRBC / 100 WBC. Since most of the auto
mated blood cell counters include NRBC in WBC count, it is necessary to subtract total NRBC
from total WBC count to get true value of total WBC count.
3) Band neutrophils (with two nucleated lobes) are counted separately. They normally
constitute less than 6% of the neutrophils. An increase in band neutrophils may be
observed in inflammatory process (in absence of an absolute leukocytosis).