Flow cytometry is a technique that uses lasers and fluorescence to count, examine, and sort cells suspended in fluid based on their optical and physical properties. Cells are stained with fluorescent markers and passed in a stream through a laser beam, which causes them to emit light at different wavelengths. Detectors then measure the light scattered and emitted to identify cell characteristics like size, granularity, and marker expression. Data is analyzed using software to identify cell populations and phenotypes. Flow cytometry has many applications in fields like immunology, hematology, and oncology for analyzing blood, tissue, and other cell samples.
FLOW CYTOMETRY, PRINCIPLE, APPLICATION, USE IN HAEMATOLOGY, COMPONENT OF FLOW CYTOMETRY, DATA INTERPRETATION, DATA ANALYSIS, CELL SHORTING ADVANTAGES AND DISADVANTAGES, IMMUNOLOGICAL CLASSIFICATION OF ACUTE
LEUKEMIA
Introduction
Definition
Basic mechanism
Prerequisite of flow cytometer
Components of flow cytometry
Flow system
Optics system
Concept of scattering
Advantage
Limitation
Application
Conclusion
References
It is a laser based technology that measures and analyses different physical and chemical properties of the cells/particles flowing in a stream of fluid through a beam of light
FLOW CYTOMETRY, PRINCIPLE, APPLICATION, USE IN HAEMATOLOGY, COMPONENT OF FLOW CYTOMETRY, DATA INTERPRETATION, DATA ANALYSIS, CELL SHORTING ADVANTAGES AND DISADVANTAGES, IMMUNOLOGICAL CLASSIFICATION OF ACUTE
LEUKEMIA
Introduction
Definition
Basic mechanism
Prerequisite of flow cytometer
Components of flow cytometry
Flow system
Optics system
Concept of scattering
Advantage
Limitation
Application
Conclusion
References
It is a laser based technology that measures and analyses different physical and chemical properties of the cells/particles flowing in a stream of fluid through a beam of light
Flow cytometry is a standard laser-based technology that is used in the detection and measurement of physical and chemical characteristics of cells or particles in a heterogeneous fluid mixture.
Flow cytometry is a standard laser-based technology that is used in the detection and measurement of physical and chemical characteristics of cells or particles in a heterogeneous fluid mixture.
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
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
- Video recording of this lecture in English language: https://youtu.be/kqbnxVAZs-0
- Video recording of this lecture in Arabic language: https://youtu.be/SINlygW1Mpc
- 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
These lecture slides, by Dr Sidra Arshad, offer a quick overview of the physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar lead (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
6. Describe the flow of current around the heart during the cardiac cycle
7. Discuss the placement and polarity of the leads of electrocardiograph
8. Describe the normal electrocardiograms recorded from the limb leads and explain the physiological basis of the different records that are obtained
9. Define mean electrical vector (axis) of the heart and give the normal range
10. Define the mean QRS vector
11. Describe the axes of leads (hexagonal reference system)
12. Comprehend the vectorial analysis of the normal ECG
13. Determine the mean electrical axis of the ventricular QRS and appreciate the mean axis deviation
14. Explain the concepts of current of injury, J point, and their significance
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. Chapter 3, Cardiology Explained, https://www.ncbi.nlm.nih.gov/books/NBK2214/
7. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Rasamanikya is a excellent preparation in the field of Rasashastra, it is used in various Kushtha Roga, Shwasa, Vicharchika, Bhagandara, Vatarakta, and Phiranga Roga. In this article Preparation& Comparative analytical profile for both Formulationon i.e Rasamanikya prepared by Kushmanda swarasa & Churnodhaka Shodita Haratala. The study aims to provide insights into the comparative efficacy and analytical aspects of these formulations for enhanced therapeutic outcomes.
Adv. biopharm. APPLICATION OF PHARMACOKINETICS : TARGETED DRUG DELIVERY SYSTEMSAkankshaAshtankar
MIP 201T & MPH 202T
ADVANCED BIOPHARMACEUTICS & PHARMACOKINETICS : UNIT 5
APPLICATION OF PHARMACOKINETICS : TARGETED DRUG DELIVERY SYSTEMS By - AKANKSHA ASHTANKAR
Local Advanced Lung Cancer: Artificial Intelligence, Synergetics, Complex Sys...Oleg Kshivets
Overall life span (LS) was 1671.7±1721.6 days and cumulative 5YS reached 62.4%, 10 years – 50.4%, 20 years – 44.6%. 94 LCP lived more than 5 years without cancer (LS=2958.6±1723.6 days), 22 – more than 10 years (LS=5571±1841.8 days). 67 LCP died because of LC (LS=471.9±344 days). AT significantly improved 5YS (68% vs. 53.7%) (P=0.028 by log-rank test). Cox modeling displayed that 5YS of LCP significantly depended on: N0-N12, T3-4, blood cell circuit, cell ratio factors (ratio between cancer cells-CC and blood cells subpopulations), LC cell dynamics, recalcification time, heparin tolerance, prothrombin index, protein, AT, procedure type (P=0.000-0.031). Neural networks, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and N0-12 (rank=1), thrombocytes/CC (rank=2), segmented neutrophils/CC (3), eosinophils/CC (4), erythrocytes/CC (5), healthy cells/CC (6), lymphocytes/CC (7), stick neutrophils/CC (8), leucocytes/CC (9), monocytes/CC (10). Correct prediction of 5YS was 100% by neural networks computing (error=0.000; area under ROC curve=1.0).
Integrating Ayurveda into Parkinson’s Management: A Holistic ApproachAyurveda ForAll
Explore the benefits of combining Ayurveda with conventional Parkinson's treatments. Learn how a holistic approach can manage symptoms, enhance well-being, and balance body energies. Discover the steps to safely integrate Ayurvedic practices into your Parkinson’s care plan, including expert guidance on diet, herbal remedies, and lifestyle modifications.
Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
1. FLOW CYTOMETRY
Subject- Pharmaceutical Biotechnology (P)
DEPARTMENT OF PHARMACEUTICAL SCIENCES
Dr. Hari Singh Gour Vishwavidyalaya
Sagar(M.P)-470003, India
(A Central University)
Submitted By:
DEBASIS SEN
(Y22254008)
Submitted To:
Prof. UMESH K. PATIL
(Professor, DOPS)
Dr. UDITAAGRAWAL
(Guest Faculty, DOPS)
Ms. POOJA DAS BIDLA
(Research Scholar, DOPS)
3. INTRODUCTION
Flow- the motion characteristics of fluids
Cyto- combining form of cells
Metry- measurement
• Hence, flow cytometry is the measurement of cell properties as cell move in a single
hydrodynamic flow and interrupt a beam of light.
• Flow cytometry is a popular cell biology technique that utilizes laser-based technology to
count, sort, and profile cells in a heterogeneous fluid mixture.
• The technique was first described by Wallace Coulter in the 1950s.
• The present flow cytometers are capable of analyzing upto 13 parametrs (forward scatter,
side scatter, 11 colours of immunofluorescence).
5. BASIC MECHANISM
Biological sample
Label it with a fluorescent marker
Cells move in a linear stream through a focused light source(laser
beam)
Fluorescent molecule gets activated and emits light that is filtered
and detected by sensitive light detectors(usually a photomultiplier
tube)
Conversion of analogue fluorescent signals to digital signal
6. SAMPLE PREPARATION
The exact steps for sample preparation can vary depending on the specific experimental design
and research question. Here is the general steps which are followed for sample preparation in
flow cytometry-
Collect the
cells or
particles of
interest
Prepare the
cells or
particles by
washing them
in a buffer
solution to
remove any
debris or
contaminants
Stain the cells
or particles
with
fluorescent
dyes or
antibodies
Fixation: In
order to stop
any further
degradation
Filtering: to
eliminate any
cellular
aggregates and
debris
Final
resuspension:
involves
resuspending
the cells or
particles in a
buffer solution
7. INSTRUMENTATION
COMPONENTS
Flow Cytometry has three main components-
Fluidics
• Transports cells in a
stream to the laser
beam for
interrogation.
Optics
• It consists of lasers
to illuminate the
particles in the
sample stream and
optical filters to
direct the resulting
light signals to the
appropriate
detectors
Electronics
• The electronics
system converts the
detected light
signals into
electronic signals
that can processed
by the computer
8.
9. FLUIDICS
The fluidic system consists of a Flow cell (Quartz chamber)
• Central core- through which the sample is injected.
• Outer sheath- contains faster flowing fluid i.e; Sheath
fluid, enclosing the central core.
• Once the sample is injected, they are forced into the
centre of the stream forming a single file.
10. OPTICS
Single wavelength, coherent light
Light Source
• Provide energy that excite the fluorochrome of interest, helps to
create stable and reliable signal.
Focus the beam
Lenses
• Keep the beam perpendicular to sample stream flow
• Make beam small enough to illuminate only one cell at a time
Filters
• Ensures that each photodetector receives light bands of various
wavelenghts
11. When a light intersects a laser beam at the so called ‘interrogation point’ two events occur-
A. Light scattering
B. Emission of fluorescent light
12. LIGHT SCATTERING
The light scatters in two ways-
• Forward Scatter (FSC)- The incident light scattered in forward direction (Along same
axis laser is travelling).
• Forward scatter represent the size of the cell.
Smaller cell will scatter less and larger will
scatter more.
13. • Side Scatter (SSC)- The incident light scattered at an angle of 90 degree to the axis of the
laser path.
• The intensity of this signal is proportional to
amount of cytosolic structure in cell (e.g-
granules, cell inclusions etc).
• Side scatter represents the granularity/ internal
complexity of cell.
14. EMISSION OF FLUORESCENT LIGHT
Fluorescence is the property of certain substances by which they absorb light at a certain wavelength
and then emit light at a longer wavelength, this substances are called fluorochromes.
Mainly used fluorochromes in flow cytometry- FITC, PE, ECD, PC5, APC, PERCP
Fluorochrome absorb some light and
become excited
Release energy in form of photons with
specific wavelength (larger than
excitation wavelength)
Photons pass through the collection lens
and splits down specific channel with the
use of filter
15. OPTICAL FILTERS
Many wavelengths of light will be scattered from a cell.
But how to split the light into its specific wavelengths to detect them independently?
Optical filters are designed such that they absorb or reflect some wavelengths of light, while
transmitting others.
Optical
Filters
Short pass
filter
Long pass
filter
Band pass
filter
Dichroic
filter
16. DETECTOR
There are mainly two types of detectors used in flow cytometry-
• Photodiode
• Photomultiplier tube (PMT)
Photodiode
• Used for strong signals
• When saturation is a potential
problem
• e.g- Forward scatter
Photomultiplier tube
(PMT)
• More sensitive and used to
detect small amounts of
fluorescence
• Band pass filter in front to
allow only specific band width
of light reach it
17. ELECTRONICS
• Detectors basically collects photos of light and convert them to current.
• The electronics process that light signal and convert the current to a digitalized value.
• This digitalized value is represented as a graph by the computer.
• Softwares-
1. Kaluza
2. CellQuest
3. FlowJo
4. WinMDI
5. FCS Express
18. DATA DISPLAY
The data are usually presented in the form of single parameter histograms or as plots or
correlated parameters, which are referred to as cytograms.
Cytograms may display data in the form of-
• Dot plot
• Density plot
• Contour plot
• Histogram
19. GATING
Gating is used to isolate a subset of cells on a plot and set a region on a histogram or
cytogram.
It allows the ability to look at parameters only on that subset.
20. COMPENSATION
Why do we need to compensate?
Fluorochromes typically give fluorescence over a large part of the spectrums (100nm or
more).
• Detectors detect fluorescence based on wavelengths
• Thus detects fluorescence from more than one fluorochrome which is called ‘Bleed Over’
• You need to compensate this bleed over so that one detector reports signals from only one
fluorochrome.
23. LIMITATIONS
• Flow cytometry is limited by its requirement that analyzed cells be in suspension, making
information on tissue architecture and cell-cell interactions unavailable.
• Cell subpopulations with similar marker expression and difficult to differentiate and
analyses that employ more fluorophores are subject to signal slipover.
• Flow cytometry may generate massive amounts of data, making analyses complicated.
24. CONCLUSION
Flow cytometry is a versatile and powerful technique that has revolutionized our
understanding of cells and their functions. Flow cytometry has a wide range of applications,
including immunology, hematology, oncology, microbiology, and stem cell research. It can
be used to analyze complex mixtures of cells, such as blood or bone marrow samples, to
identify and quantify different cell types. Flow cytometry can also be used to study the cell
cycle, apoptosis, and cell signaling pathways. So it has widespread use in research and
clinical settings has led to numerous discoveries and advancements in medicine and biology.
25. REFERENCES
1. Büscher M. Flow cytometry instrumentation–an overview. Current Protocols in
Cytometry. 2019 Jan;87(1):e52.
2. Nunez R. Flow cytometry: principles and instrumentation. Current Issues in Molecular
Biology. 2001 Apr;3(2):39-45.
3. Hoffman RA. Flow cytometry: instrumentation, applications, future trends and
limitations. Standardization and Quality Assurance in Fluorescence Measurements II:
Bioanalytical and Biomedical Applications. 2008:307-42.
4. Chapman GV. Instrumentation for flow cytometry. Journal of immunological methods.
2000 Sep 21;243(1-2):3-12.