Flow cytometry uses laser-based technology to identify and quantify cell populations in a fluid suspension. Cells flow through the system and are interrogated by detectors that measure their physical properties like size and complexity. It can identify normal vs abnormal cells, differentiate cell types, and quantify tumor infiltration. Key components include lasers, fluidics to align cells, optical systems to filter light signals, and detectors to convert fluorescence into electrical signals for analysis. It allows multiparameter analysis using antibody panels to detect cell surface and intracellular antigens, aiding diagnosis and monitoring of hematological malignancies.
A brief presentation on cell counting and cell viability assays. For cell cytotoxicity assays, you can check my profile where I have uploaded a separate file.
Prepared in July 2015
A brief presentation on cell counting and cell viability assays. For cell cytotoxicity assays, you can check my profile where I have uploaded a separate file.
Prepared in July 2015
This work is done in IIT-M (Indian Institute of Technology- Madras) with help of Indian Academy of Science during June 2011-Oct 2011 under Dr Karunagaran Devarajan sir
Explain the basic mechanisms involved in DNA extraction.
Describe the steps involved in gDNA extraction from blood.
Explain the processes involved in quality and quantity check of extracted DNA using nanodrop technique.
Decribe the steps of quantity check of amplicon using flurometer.
Decribe the principle of dilution of amplicon.
Presented by,
Dr. Md. Mohiuddin Masum
Guided by,
Prof. Laila Anjuman Banu
The extraction of DNA involves three main steps that are cell lysis, protein separation, and DNA purification. Cell lysis is usually performed by incubation of cell in buffer containing detergent and protease. Cellular proteins are salted out or phase separated using organic solvents. Finally DNA is isolated and purified either by alcohol precipitation or adsorption with silica and elution.
Define karyotype and FISH
Describe the procedure of karyotyping and FISH
Explain chromosomal abnormalities through karyotyping and FISH
Describe the principles of FISH
Presented by-
Dr. Subarna Das
Resident, MS Anatomy
Phase-A, Year-1, Block-2
Guided by-
Prof. Laila Anjuman Banu
Chairman
Department of Anatomy, BSMMU
This work is done in IIT-M (Indian Institute of Technology- Madras) with help of Indian Academy of Science during June 2011-Oct 2011 under Dr Karunagaran Devarajan sir
Explain the basic mechanisms involved in DNA extraction.
Describe the steps involved in gDNA extraction from blood.
Explain the processes involved in quality and quantity check of extracted DNA using nanodrop technique.
Decribe the steps of quantity check of amplicon using flurometer.
Decribe the principle of dilution of amplicon.
Presented by,
Dr. Md. Mohiuddin Masum
Guided by,
Prof. Laila Anjuman Banu
The extraction of DNA involves three main steps that are cell lysis, protein separation, and DNA purification. Cell lysis is usually performed by incubation of cell in buffer containing detergent and protease. Cellular proteins are salted out or phase separated using organic solvents. Finally DNA is isolated and purified either by alcohol precipitation or adsorption with silica and elution.
Define karyotype and FISH
Describe the procedure of karyotyping and FISH
Explain chromosomal abnormalities through karyotyping and FISH
Describe the principles of FISH
Presented by-
Dr. Subarna Das
Resident, MS Anatomy
Phase-A, Year-1, Block-2
Guided by-
Prof. Laila Anjuman Banu
Chairman
Department of Anatomy, BSMMU
An array of presentation of lymphoma spillover in the peripheral smear and bone marrow. All types of lymphomas are discussed along with a bouquet of HPE pictures
Discussion on Papillary carcinoma thyroid - category, types with definitions, other DD for pap ca thyroid along with a whole lot of beautiful cytology pictures - a feast to the eyes
Discussion on types, pathogenesis of hypersensitivity. Referred from Robbins 10th edition. Prepared by a pathology Postgraduate for 2nd year MBBS students
263778731218 Abortion Clinic /Pills In Harare ,sisternakatoto
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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
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
The prostate is an exocrine gland of the male mammalian reproductive system
It is a walnut-sized gland that forms part of the male reproductive system and is located in front of the rectum and just below the urinary bladder
Function is to store and secrete a clear, slightly alkaline fluid that constitutes 10-30% of the volume of the seminal fluid that along with the spermatozoa, constitutes semen
A healthy human prostate measures (4cm-vertical, by 3cm-horizontal, 2cm ant-post ).
It surrounds the urethra just below the urinary bladder. It has anterior, median, posterior and two lateral lobes
It’s work is regulated by androgens which are responsible for male sex characteristics
Generalised disease of the prostate due to hormonal derangement which leads to non malignant enlargement of the gland (increase in the number of epithelial cells and stromal tissue)to cause compression of the urethra leading to symptoms (LUTS
Acute scrotum is a general term referring to an emergency condition affecting the contents or the wall of the scrotum.
There are a number of conditions that present acutely, predominantly with pain and/or swelling
A careful and detailed history and examination, and in some cases, investigations allow differentiation between these diagnoses. A prompt diagnosis is essential as the patient may require urgent surgical intervention
Testicular torsion refers to twisting of the spermatic cord, causing ischaemia of the testicle.
Testicular torsion results from inadequate fixation of the testis to the tunica vaginalis producing ischemia from reduced arterial inflow and venous outflow obstruction.
The prevalence of testicular torsion in adult patients hospitalized with acute scrotal pain is approximately 25 to 50 percent
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
New Drug Discovery and Development .....NEHA GUPTA
The "New Drug Discovery and Development" process involves the identification, design, testing, and manufacturing of novel pharmaceutical compounds with the aim of introducing new and improved treatments for various medical conditions. This comprehensive endeavor encompasses various stages, including target identification, preclinical studies, clinical trials, regulatory approval, and post-market surveillance. It involves multidisciplinary collaboration among scientists, researchers, clinicians, regulatory experts, and pharmaceutical companies to bring innovative therapies to market and address unmet medical needs.
NVBDCP.pptx Nation vector borne disease control programSapna Thakur
NVBDCP was launched in 2003-2004 . Vector-Borne Disease: Disease that results from an infection transmitted to humans and other animals by blood-feeding arthropods, such as mosquitoes, ticks, and fleas. Examples of vector-borne diseases include Dengue fever, West Nile Virus, Lyme disease, and malaria.
Title: Sense of Smell
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the primary categories of smells and the concept of odor blindness.
Explain the structure and location of the olfactory membrane and mucosa, including the types and roles of cells involved in olfaction.
Describe the pathway and mechanisms of olfactory signal transmission from the olfactory receptors to the brain.
Illustrate the biochemical cascade triggered by odorant binding to olfactory receptors, including the role of G-proteins and second messengers in generating an action potential.
Identify different types of olfactory disorders such as anosmia, hyposmia, hyperosmia, and dysosmia, including their potential causes.
Key Topics:
Olfactory Genes:
3% of the human genome accounts for olfactory genes.
400 genes for odorant receptors.
Olfactory Membrane:
Located in the superior part of the nasal cavity.
Medially: Folds downward along the superior septum.
Laterally: Folds over the superior turbinate and upper surface of the middle turbinate.
Total surface area: 5-10 square centimeters.
Olfactory Mucosa:
Olfactory Cells: Bipolar nerve cells derived from the CNS (100 million), with 4-25 olfactory cilia per cell.
Sustentacular Cells: Produce mucus and maintain ionic and molecular environment.
Basal Cells: Replace worn-out olfactory cells with an average lifespan of 1-2 months.
Bowman’s Gland: Secretes mucus.
Stimulation of Olfactory Cells:
Odorant dissolves in mucus and attaches to receptors on olfactory cilia.
Involves a cascade effect through G-proteins and second messengers, leading to depolarization and action potential generation in the olfactory nerve.
Quality of a Good Odorant:
Small (3-20 Carbon atoms), volatile, water-soluble, and lipid-soluble.
Facilitated by odorant-binding proteins in mucus.
Membrane Potential and Action Potential:
Resting membrane potential: -55mV.
Action potential frequency in the olfactory nerve increases with odorant strength.
Adaptation Towards the Sense of Smell:
Rapid adaptation within the first second, with further slow adaptation.
Psychological adaptation greater than receptor adaptation, involving feedback inhibition from the central nervous system.
Primary Sensations of Smell:
Camphoraceous, Musky, Floral, Pepperminty, Ethereal, Pungent, Putrid.
Odor Detection Threshold:
Examples: Hydrogen sulfide (0.0005 ppm), Methyl-mercaptan (0.002 ppm).
Some toxic substances are odorless at lethal concentrations.
Characteristics of Smell:
Odor blindness for single substances due to lack of appropriate receptor protein.
Behavioral and emotional influences of smell.
Transmission of Olfactory Signals:
From olfactory cells to glomeruli in the olfactory bulb, involving lateral inhibition.
Primitive, less old, and new olfactory systems with different path
2. INTRODUCTION
• Laser – based technology
• Identify & quantify cell populations
• Cells are manipulated into a stream of fluid
• Interrogated by an electronic detection system
Flow - Fluid
Cyto - Cell
Metry –
Measurement
Measures properties of the cell as they flow in a fluid
suspension across an illuminated light path
4. SAMPLE PREPARATION
• Peripheral blood
• Bone marrow
• CSF
• Ascitic fluid
• Pleural fluid
• FNA
• Red cells are lysed
• Appropriate antibody panels
are chosen
Spleen
Lymphnode
Liver
Bone marrow biopsy
After tissue disaggregation
5. USES
• Identification & quantification of cell populations within a sample
• Normal vs abnormal cells
• Reactive vs neoplastic cells
• Differentiation & maturation stage in a cell population
• Quantification of tumour infiltration
6. COMPONENTS
• Laser
• Argon (most common)
• Monochromatic light
• Sheath fluid
• Aligns cells in a single file
• Optical systems
• Filter and regulate light signals
• Photomultiplier detectors
• forward light scatter – cell size
• Side light scatter – cell complexity
• Converting fluorescent light signal into electrical signal
• Computer
Fluidics -
Hydrodynamic focusing
Excitation Optics &
Collection Optics
Electronics
7.
8. How does single filing occur?
• Hydrodynamic focusing
• Sheath fluid exerts pressure onto the sample
• As the nozzle narrows -> a single file is obtained
9. Forward Scatter & Side Scatter
• Forward Scatter – Proportional to the size of the cell
• Side Scatter – proportional to the Complexity of the cell
20. • STAIN-LYSE-WASH METHOD
• Dilute cell concentration to 1 – 2 x 106 per tube
Pipette 100 μl specimen in round bottom tube +
McAb combination or Multicolor cocktail
Incubate at room temperature in dark room for 15
min
1 ml of NH4Cl based lysing solution &
Incubate for 10 mins
Centrifuge x 5 mins at 300 g
Discard the supernatant & repeat
Resuspend cells in 0.2 – 0.5 ml of sheath fluid
21. Stain-lyse-no wash method
• No centrifugation
• Rest of the steps are same
• Ideal for samples with few cells
• Cell loss during centrifugation is minimised
22. Lyse-stain-wash method
• Bulk
specimen
lysis
• Used for
MRD
monitoring
5 – 10 ml of sample
+ same amount of
NH4Cl lysing
solution
Mix gently &
incubate at room
temperature for 10
mins
Centrifuge at 300g
for 5 min
Discard supernatant
Resuspend cell
pellet in 10 ml PBS-
Azide-BSA
Repeat washing &
resuspend
Aliquot a volume of
cell suspension
containing 10 x 106
cells/tube
Add McAb
Incubate in dark
Repeat washing &
resuspend in 0.2 –
0.5 ml of sheath
fluid
Interpret data
23. Detection of intracellular antigens
• Fix & Perm kit
• Solution A – Fixing agent (paraformaldehyde solution)
• Solution B – lysing agent (lysing solution & detergent)
Pipette 100 μl
of specimen
Add 100 μl of
Solution A &
Incubate at
room temp x
15 mins
Wash twice in
PBS-Azide-
BSA
Add 100 μl of
Solution B +
Ab cocktail
Incubate at
room
temperature
in dark x 15
mins
Wash twice
with PBS-
Azide-BSA
Resuspend in
sheath fluid
Interpret Data
25. MULTICOLOR OR MULTIPARAMETER FLOW
CYTOMETRY
• 8 – 10 color antibody panels
• Provides data of 12 – 14 cellular parameters simultaneously
ADVANTAGES DISADVANTAGES
Increased accuracy Increased complexity of compensation
Smaller sample size Challenges of antibody panel validation
Cost effectiveness Tandem dye conjugate issues
Increased efficiency Increased need for expertise in data
analysis & interpretation
Increased sensitivity for minimal residual
disease
Human error
