Research_Methods.pptx

Research
Methods
Basic Laboratory Techniques and
Methods
Presented by: HANNAH B. DELOS REYES

Review
Let’s have a short recap of
what we had discussed last
meeting!

Review
What are the two (2) general
types of sampling methods?

Review
Can you give one example
of scientific sampling
method and explain it?

Review
Can you give one example of
non-scientific sampling
method and explain it?

Objective
s
At the end of the lesson, the
learners should be able to:

At the end of the lesson, the
learners should be able to:
Identify standard
methods and
techniques use in
performing
experiments.
Describe proper
use of specific,
standard
laboratory
procedure.
Enumerate basic
standard methods
and techniques
use in performing
experiments.
Apply proper
handling and
setting of
laboratory
equipment.

- Biological Methods
- Chemical Methods
- Physical Methods
Research Methods

- Microscopy
- Tissue Culture
- Aseptic Technique
- Dissection
- Agarose Gel Electrophoresis
- Polymerase Chain Reaction (PCR)
Biological Methods

Microscopy is the technical field of using
microscopes to view objects and areas of
objects that cannot be seen with the naked eye
(objects that are not within the resolution
range of the normal eye).
Microscopy

There are three well-known branches of
microscopy: optical, electron and scanning
probe, along with the emerging field of X-ray
microscopy.
Microscopy

Optical microscopes bounce light off the thing
you’re trying to view.
Electron microscopy creates an image by firing
electrons at an object.
Microscopy

Scanning probe microscopy involves running
a physical probe across the surface of the
material. It’s the equivalent of a blind person
feeling their way around an object.
Microscopy

- Analysis of the distribution of
biochemical components (proteins,
lipids, sugars, etc.) inside a tissue
section (animal or plant material)
- Another application of electron
microscopy is forensic science
- Characterization and classification of
tissues
Applications

- Differentiation of tumors
- Diagnostics of diseases by pattern
recognition
- Microscopy is used to diagnose the
illness in the hospitals and the clinics
all over the world
Applications

- Microscopes help the scientists to
study the microorganisms, the cells,
the crystalline structures and the
molecular structures
- Have opened up a whole new
dimension in science
- Help create the very tiny electrical
circuits found on Silicon microchips
Importance

- Without microscope we hardly be
able to identify, study or see many
multicellular, unicellular organisms
their body parts, cells & tissues,
molecular structures of DNA, water,
diamond, glucose etc.
Importance

- Microscopes enlarge the view of tiny
organisms which allows scientists to
develop the vaccine & cures for
infectious diseases in the humans and
the animals.
Importance

In biological research, tissue culture
refers to a method in which fragments of a
tissue (plant or animal tissue) are introduced
into a new, artificial environment, where they
continue to function or grow.
Tissue Culture

- Virus reproduction
- Micro propagation of plants
- Growing human tissue for organ
transplants
- Producing biotechnology products
- Biomass Energy
- Secondary Metabolites
Application

- Genetic Variability
- Somatic Embryogenesis and Synthetic
Seed
- Breaking Dormancy
- Haploid Plants and Somatic Hybrids
- Transgenic Plants
- Germplasm Conservation
Application

Aseptic technique means using practices and
procedures to prevent contamination from
pathogens. It involves applying the strictest
rules to minimize the risk of infection.
Aseptic Techniques

- Handling surgery equipment
- Helping with a baby’s birth by vaginal
delivery
- Handling dialysis catheters
- Performing dialysis
- Inserting a chest tube
Application

- Inserting a urinary catheter
- Inserting central intravenous (iv) or
arterial lines
- Inserting other draining devices
- Performing various surgical
techniques
Application

- To protect patients from harmful
bacteria and other pathogens during
medical procedures, healthcare
providers use aseptic technique.
- Proper aseptic technique prevents
contamination of cultures from foreign
bacteria inherent in the environment.
Importance

Dissection is the process of cutting apart or
separating tissue as, for example, in the study
of anatomy or in the course of a surgical
procedure.
Dissection

- Determine the cause of death in
autopsy
- An intrinsic part of forensic
- Has been used for centuries to
explore anatomy
Application

Agarose electrophoresis is a technique used to
separate DNA fragments based on their size
involves running a current through a gel
containing the molecules of interest.
Agarose Gel
Electrophoresis

The PCR is a scientific technique in molecular
biology to amplify a single or a few copies of a
piece of DNA across several orders of
magnitude, generating thousands to millions of
copies of a particular DNA sequence.
Polymerase Chain Reaction

- Genetic testing
- Tissue typing prior to organ
transplantation
- Formulation of individualized cancer
therapy treatments
- Genetic fingerprinting
- Parental testing
Application

- Early detection of several forms of
cancer including leukemia and
lymphoma.
- Detection of viral DNA and virulent
sub-types
- DNA sequencing, DNA cloning and
gene expression
- Production of hybridization probes
Application

- It can be used to amplify a sample of
DNA when there isn’t enough to
analyze.
- As a method of identifying a gene of
interest.
- PCR can detect and identify bacteria
and viruses that cause infections.
Importance

- PCR is also used in genetic testing, to
determine whether patients carry a
genetic mutation that could be passed
on to their or to determine disease risk
in patients themselves.
- Scientists can gather information
about evolutionary relationships using
PCR on ancient samples.
Importance

- Spectrophotometry
- Extraction
- Titration
- Qualitative Test for Secondary Metabolites
Chemical Methods

Spectrophotometry is a method to measure
how much a chemical substance absorbs light
by measuring the intensity of light as a beam
of light passes through sample solution.
Spectrophotometry

- Measure the concentration of a
solution
- Identify organic compounds by
determining the absorption in
maximum
- Used for color determination within
the spectral range
Uses

- Forensic sciences
- Molecular biology
- In disease diagnosis/ pathological
states (changes)
- Blood (the blood plasma, and the
formed elements
Applications

- To identify and quantify biological
molecules
- Measurement of concentration
- Detection of impurities
- Elucidation of the structure of
organic compounds
- Chemical kinetics
Importance

Extractions are a way to separate a desired
substance when it is mixed with others. The
mixture is brought into contact with a solvent
in which the substance of interest is soluble,
but the other substances present are insoluble.
Extraction

Maceration
This is a very simple extraction method with
the disadvantage of long extraction time and
low extraction efficiency.
Extraction

Percolation
Percolation is more efficient than maceration
because it is a continuous process in which
the saturated solvent is constantly being
replaced by fresh solvent.
Extraction

Decoction
The extract from decoction contains a large
amount of water-soluble impurities. Decoction
cannot be used for the extraction of
thermolabile or volatile components.
Extraction

Reflux Extraction
Reflux extraction is more efficient than
percolation or maceration and requires less
extraction time and solvent. It cannot be used
for the extraction of thermolabile natural
products.
Extraction

Soxhlet Extraction
The Soxhlet extraction method integrates the
advantages of the reflux extraction and
percolation, which utilizes the principle of reflux
and siphoning to continuously extract the herb
with fresh solvent.
Extraction

Pressurized Liquid Extraction (PLE)
PLE has also been described as accelerated
solvent extraction, enhanced solvent extraction,
pressurized fluid extraction, accelerated fluid
extraction, and high pressure solvent extraction.
Extraction

Supercritical Fluid Extraction (SFE)
Supercritical fluid extraction (SFE) uses
supercritical fluid (SF) as the extraction solvent.
SF has similar solubility to liquid and similar
diffusivity to gas, and can dissolve a wide variety
of natural products.
Extraction

Ultrasound Assisted Extraction (UAE)
Ultrasonic-assisted extraction (UAE), also
called ultrasonic extraction or sonication, uses
ultrasonic wave energy in the extraction.
Extraction

Pulsed Electric Field (PEF) Extraction
Pulsed electric field extraction significantly
increases the extraction yield and decreased the
extraction time because it can increase mass
transfer during extraction by destroying
membrane structures.
Extraction

Enzyme assisted extraction (EAE)
The extraction efficiency will be enhanced by EAE
due to the hydrolytic action of the enzymes on the
components of the cell wall and membrane and the
macromolecules which facilitate the release of the
natural product.
Extraction

Research_Methods.pptx

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