Centrifugation principle and types by Dr. Anurag YadavDr Anurag Yadav
concept of cnetrifugation,
basic Principle
centrifugal force
types of centrifugation based on use and rotor type
application of the each type of centrifuge
Ultracentrifuge in detail
application in general
Centrifugation is the separation technique commonly used in clinical and research laboratories.
It is based on the behavior of particles in an applied centrifugal field.
More dense components of the mixture move away from the axis of the centrifuge while less dense components of the mixture move towards the axis.
Centrifugation principle and types by Dr. Anurag YadavDr Anurag Yadav
concept of cnetrifugation,
basic Principle
centrifugal force
types of centrifugation based on use and rotor type
application of the each type of centrifuge
Ultracentrifuge in detail
application in general
Centrifugation is the separation technique commonly used in clinical and research laboratories.
It is based on the behavior of particles in an applied centrifugal field.
More dense components of the mixture move away from the axis of the centrifuge while less dense components of the mixture move towards the axis.
Centrifugation is a mechanical process which involves the use of the centrifugal force to separate particles from a solution according to their size, shape, density, medium viscosity and rotor speed. The larger the size and the larger the density of the particles, the faster they separate from the mixture.
A biochemical technique used for isolation of molecules on the basis of size ,shape & density. Categorized into two types:
1) Analytical ultracentrifugation
2) Preparative ultracentrifugation
It is an important tool in biochemical research. Which through rapid spinning imposes high centrifugal forces on suspended particles, or even molecules in solution, and causes separations of such matter on the basis of differences in weight.
Bright field microscopy, Principle and applicationsKAUSHAL SAHU
Introduction
History
Basic Component of Microscope
Light Microscopy
Types of Light Microscopy
What Are Bright Microscopy
Principle of Bright Microscope
Advantage
Disadvantage
Application
Conclusion
Reference
A spectrophotometer is an instrument containing a monochromator, a device which produces a light beam containing wavelengths in a narrow band around a selected wavelength, and a means of measuring the ratio of that beam's intensity as it enters and leaves a cuvette 99 This describes a single-beam photometer.
Centrifugation is a mechanical process which involves the use of the centrifugal force to separate particles from a solution according to their size, shape, density, medium viscosity and rotor speed. The larger the size and the larger the density of the particles, the faster they separate from the mixture.
A biochemical technique used for isolation of molecules on the basis of size ,shape & density. Categorized into two types:
1) Analytical ultracentrifugation
2) Preparative ultracentrifugation
It is an important tool in biochemical research. Which through rapid spinning imposes high centrifugal forces on suspended particles, or even molecules in solution, and causes separations of such matter on the basis of differences in weight.
Bright field microscopy, Principle and applicationsKAUSHAL SAHU
Introduction
History
Basic Component of Microscope
Light Microscopy
Types of Light Microscopy
What Are Bright Microscopy
Principle of Bright Microscope
Advantage
Disadvantage
Application
Conclusion
Reference
A spectrophotometer is an instrument containing a monochromator, a device which produces a light beam containing wavelengths in a narrow band around a selected wavelength, and a means of measuring the ratio of that beam's intensity as it enters and leaves a cuvette 99 This describes a single-beam photometer.
Centrifugation is a process which involves the use of the centrifugal force for the sedimentation of heterogeneous mixtures with centrifuge used in laboratory. If the particle size in dispersion is 5 micro meter or less then they undergo Brownian motion hence don't sediment under gravity. Therefore a stronger force , centrifugal force is applied to separate the suspended particles.
Basics of DNA isolation, What is chemistry behind it. Presently the laboratory of animal science department ,Göttingen university using this technique for dna isolation in pig blood sample.
A centrifuge operates by using the sedimentation principle- Here the substances are separated based on their density under the influence of gravitational force. When spun rapidly, lighter particles stay at the top and heavier particles go to the bottom during centrifugation.The components of heterogeneous mixtures are detached by centrifugation. That comprises liquids in liquids, solids in fluids, and gases in solids and liquids. In order to transfer bulky sections to the outside of the pipe, centrifugation uses centrifugal energy. It allows the solid to settle more easily and completely.The components of heterogeneous mixtures are detached by centrifugation. That comprises liquids in liquids, solids in fluids, and gases in solids and liquids. In order to transfer bulky sections to the outside of the pipe, centrifugation uses centrifugal energy. It allows the solid to settle more easily and completely.
What is the centrifuge used for?
Centrifuges work by separating out two materials with different densities. Centrifuges are used in various laboratories to separate fluids, gases or liquids based on density like the separation of different constituents of blood, immiscible liquids, wastewater sludge etc.
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We understand the unique challenges pickleball players face and are committed to helping you stay healthy and active. In this presentation, we’ll explore the three most common pickleball injuries and provide strategies for prevention and treatment.
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Growing Prevalence of Lifestyle Diseases
The rising incidence of lifestyle diseases such as diabetes, cardiovascular diseases, and cancer is a major trend driving the clinical trials market in India. These conditions necessitate the development and testing of new treatment methods, creating a robust demand for clinical trials. The increasing burden of these diseases highlights the need for innovative therapies and underscores the importance of India as a key player in global clinical research.
CHAPTER 1 SEMESTER V - ROLE OF PEADIATRIC NURSE.pdfSachin Sharma
Pediatric nurses play a vital role in the health and well-being of children. Their responsibilities are wide-ranging, and their objectives can be categorized into several key areas:
1. Direct Patient Care:
Objective: Provide comprehensive and compassionate care to infants, children, and adolescents in various healthcare settings (hospitals, clinics, etc.).
This includes tasks like:
Monitoring vital signs and physical condition.
Administering medications and treatments.
Performing procedures as directed by doctors.
Assisting with daily living activities (bathing, feeding).
Providing emotional support and pain management.
2. Health Promotion and Education:
Objective: Promote healthy behaviors and educate children, families, and communities about preventive healthcare.
This includes tasks like:
Administering vaccinations.
Providing education on nutrition, hygiene, and development.
Offering breastfeeding and childbirth support.
Counseling families on safety and injury prevention.
3. Collaboration and Advocacy:
Objective: Collaborate effectively with doctors, social workers, therapists, and other healthcare professionals to ensure coordinated care for children.
Objective: Advocate for the rights and best interests of their patients, especially when children cannot speak for themselves.
This includes tasks like:
Communicating effectively with healthcare teams.
Identifying and addressing potential risks to child welfare.
Educating families about their child's condition and treatment options.
4. Professional Development and Research:
Objective: Stay up-to-date on the latest advancements in pediatric healthcare through continuing education and research.
Objective: Contribute to improving the quality of care for children by participating in research initiatives.
This includes tasks like:
Attending workshops and conferences on pediatric nursing.
Participating in clinical trials related to child health.
Implementing evidence-based practices into their daily routines.
By fulfilling these objectives, pediatric nurses play a crucial role in ensuring the optimal health and well-being of children throughout all stages of their development.
Leading the Way in Nephrology: Dr. David Greene's Work with Stem Cells for Ki...Dr. David Greene Arizona
As we watch Dr. Greene's continued efforts and research in Arizona, it's clear that stem cell therapy holds a promising key to unlocking new doors in the treatment of kidney disease. With each study and trial, we step closer to a world where kidney disease is no longer a life sentence but a treatable condition, thanks to pioneers like Dr. David Greene.
How many patients does case series should have In comparison to case reports.pdfpubrica101
Pubrica’s team of researchers and writers create scientific and medical research articles, which may be important resources for authors and practitioners. Pubrica medical writers assist you in creating and revising the introduction by alerting the reader to gaps in the chosen study subject. Our professionals understand the order in which the hypothesis topic is followed by the broad subject, the issue, and the backdrop.
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1. PRINCIPLES AND APPLICATIONS OF
CENTRIFUGATION
Prakash
B.Sc MLT Ist year
Department of Biochemistry
Post Graduate Institute of Medical Education And Research, Chandigarh
Moderated by:
Respected Dinesh Sir
2. INTRODUCTION
WHAT IS CENTRIFUGE?
Centrifuge is a device for separating particles from a solution according to there size,
shape, density, viscosity of the medium.
WHAT IS CENTRIFUGATION?
Centrifugation is a process which involves the use of the centrifugal force for the
sedimentation of heterogeneous mixtures with a centrifuge.
3. Centrifugal force, word from Latin Centrum, meaning “centre”, and fugere, means
“to flee”, is the apparent force that draws a rotating body away from the centre of
rotation.
The term centrifugal force is used to refer to one of two distinct concepts: an
inertial force observed in a non-inertial reference frame, and a reaction force
corresponding to a centripetal force.
The concept of centrifugal force is applied in rotating devices such as centrifuges,
centrifugal pumps. The two different forces are equal in magnitude, but centrifugal
forces is opposite in direction to the centripetal force.
Centrifugal force
4. Principle
The centrifuge involves principle of centrifugation, where the
acceleration at centripetal force causes denser substances to separate out
along the radial direction at the bottom of the tube.
In a solution, particles whose density is higher than that of the solvent
sink (sediment), and particles that are lighter than it float to the top.
The greater the difference in density, the faster they move.
5. Relative Centrifugal Force (RCF)
RCF, is the ratio of the centrifugal acceleration at a specified radius and the
speed to the standard acceleration of gravity.
Relative Centrifugal force is defined as f=Mω2 r
Where,
F= intensity of centrifugal force
M= mass of particle
ω= angular velocity of rotation
R= distance of migrating particles from central axis of rotation.
6. The RCF value for any centrifuge may be calculated from the
following equation: RCF = 1.18 × r × (rpm) 2 ×10-5
Where,
1.18×10-5 = An empirical factor
r is the radius in cm from the centre of the centrifuge shaft to the
external tip of centrifuge tube.
rpm is the number of revolution per minute of the centrifuge rotor
7. The time required to separate particles depends on the rotor speed, radius of rotor
and effective path length travelled by sedimented particles.
The following formula useful for the calculation of the speed required for a rotor
with a radius that differ from the radius with which a prescribed,
R.P.M. =1000× √RCF
1.18×r
The length of time of centrifugation also can be calculated so, that running with an
alternative rotor of a different size is equivalent to running with a original rotor.
8. Types of Centrifuge depends on :
Speed of sedimentation (ultra centrifuge or high speed centrifuge).
Presence /absence of vacuum (ultra centrifuge or small bench top)
Temperature control refrigeration.
Volume of sample and capacity of centrifugation tubes
9. Depending on the particular
application, centrifuges differ in
their overall design and size.
A common feature in all
centrifuges is the central motor
that spins a rotor containing the
samples to be separated.
10. With or without refrigeration
Slow speed (eg up to 4000
RPM)
Common in clinical lab
(blood/plasma/serum separation)
Can take approx (up to) 100
tubes, depending on diameter
Small Bench top
11. Micro centrifuge (“microfuge”,
“Eppendrof”)
Sample volume is small in
eppendrof tubes
Refrigerated with or without
Centrifuge maximum approx
10000 g
Take tube of small volume
up to 2ml.
Commonly used of
concentration protein
12. High speed centrifuge
Refrigerated
Use for protein precipitates, large intact
organelles cellular debris from tissue
homogenization and microorganism
They operate maximal centrifugal force of
approx 50000g
Use for research applications
Differential separation of nucleus,
mitochondrial, protein precipitate, etc.
13. Ultra centrifuge
Refrigerated and evacuated
The detail biochemistry analysis
of subcellular structures and isolate
biomolecules.
Operate at upto 90000 g
15. Fixed angle rotor
Ideally tool for pelleting
Isopycnic banding may form
Centrifugation tubes are held at at fixed angle of between
14'-40' to vertical axis of rotation.
Start of centrifugation particles are driven outward
horizontally but strike side of the tube so that
sediment pack against the side & bottom of the tube,
with the surface of the sediment parallel to the shaft
of centrifuge.
16.
17. Swinging bucket rotors:
Sample tubes are loaded into individual buckets that hang vertically while
the rotor is at rest.
When the rotor begins to rotate the buckets swing out to a horizontal
position. Useful when samples are to be resolved in density gradients.
The longer path length permits better separation of individual particle types
from a mixture.
This rotor is relatively inefficient for pelleting .
20. Verification of rotation speed:
Centrifuges used in the laboratory are to be considered as contaminated and
should only be handled with gloves and other personal protective equipment
and/or thoroughly disinfected before calibration verification.
Place a small section of black and white reflective tape that comes with the
tachometer on the center spindle of the test centrifuge.
In order to measure the rotation speed, there must be a viewing port in the top
cover that will allow the tachometer line of sight to the reflective tape.
Place a normal well-balanced load using specimen covers into the centrifuge.
Start the centrifuge and allow it come to equilibrium at a normal operating
speed.
Use the tachometer through the viewing port above the reflective tape to take
a rotation rate reading.
Record the rotation speed indicated by the centrifuge either by the dial setting
or by a built in tachometer on the centrifuge .
21. Verification of timer :
Centrifuges used in the laboratory are to be considered as
contaminated and should only be handled with gloves and other
personal protective equipment and/or thoroughly disinfected before
calibration verification.
Set the centrifuge timer at a setting frequently used in procedures, and
start the stopwatch simultaneously.
Stop the stopwatch at the same time as the centrifuge timer ends.
Calculate the difference between the two times .
22. Interpretation of results:
Verification of rotation speed:
If the difference between the test centrifuge and the certified tachometer
is ± 5% of the procedure speed, then the test centrifuge rotation
calibration is verified as acceptable.
Verification of timer:
Acceptable difference between the test timer and the certified timer must
be ±2% of the total test timer setting.
23. SAFETY MEASURES
The work surface must be level and firm.
Balance the tubes in the rotor.
Do not open the lid while the rotor is moving.
Bucket is properly cushioned.
If you see it wobbling or shaking, turn it off or pull the plug. A little vibration is normal, but
excessive amounts can mean danger.
Wear a face shield and / or safety goggles if you have to work anywhere near a centrifuge
that’s in use.
Examine tubes and bottles for cracks or stress marks before using them.
Discard any centrifuge tubes that have cracks in them
Clean the rotor and centrifuge well after each use
24. Maintenance of Centrifuge
Daily maintenance
Wipe the inside of the bowl with disinfectant solution and rinse thoroughly.
The centrifuge must not be used if the interior is hot, if unusual vibrations or noises occur, or if
deterioration (corrosion of parts) is detected.
A qualified service technician should be contacted.
Most vibrations are due to improper balancing and can be corrected by rebalancing the buckets and
tubes.
Monthly maintenance
Clean the centrifuge housing, rotor chamber, rotors and rotor accessories with a neutral cleaning
agent.
Clean plastic and non-metal parts with a fresh solution of 0.5% sodium hypochlorite .
25. Application
Remove cellular elements from blood to provides cell free serum or
plasma
Conc. Cellular element for microscopy.
Remove protein precipitate from analytic sample.
Isolation of macromolecules such as DNA, RNA, proteins, or lipids
Use in haematology lab for PCV determination.
26. Reference
Principles and technique in biochemistry-L WALKER&
WILSON
Lynch's Medical Laboratory Technology (4th edition)