HEMA INSTRUMENTATION Flashcards
AUTOMATION IN HEMATOLOGY
- Cell Counts (Automated Hematology Analyzers)
- Diagnosis of Hemoglobinopathies (HPLC)
- Immunophenotyping (Flow Cytometry)
- Coagulation (Coagulometers)
types of automated techniques of blood counting:
- Semi-automated Instruments
- Fully-automated Instruments
- Require some steps, as dilution of blood samples. Often measure only a small number of variables.
- Some steps like dilution of blood is carried out
manually. - Measures only a few parameter.
Semi-automated instruments
in semi-automated instruments, it require some steps, as dilution of blood samples. Often measure only a small number of ______.
variables
in semi-automated instruments, some steps like dilution of blood is carried out _______
manually
in semi-automated instruments, measures only a few _______
parameter
o Require only that an appropriate blood
sample is presented to the instrument.
o They can measure 8-20 variables including
some new parameters which do not have any equivalent in manual methods.
o Measures multiple parameters
o Requires only anticoagulated blood samples.
Fully automated instruments
Fully automated instruments, require only that an appropriate _________ is presented to the __________.
blood sample ; instrument
Fully automated instruments, they can measure _____ variables including some _________ which do not have any equivalent in manual methods.
8-20 ; new parameters
Fully automated instruments, measures __________
multiple parameters
Fully automated instruments, requires only ______________
anticoagulated blood samples
Cost effective and convenient and Several CBCs performed daily
Automated Analyzers
Benefits of Automated Analyzers
o Reduced labor costs
o More complete information
o Improved data reliability
FEATURES OF PENTRA DF NEXUS
- Methods of measurement: cytochemistry,
impedance and flow cytometry. - Balance concept: automatic control of the
leucocyte count based on 3 independent
principles. - DHSS: focused flow cytometry and sequential
measurement (impedance and absorbance). - Automatic reflex testing: selective and
programmable (hematology parameters, alarms
and flags, demography). - Rack rotation mixing of samples: smooth and
efficient - Quality: complete traceability for each run in
agreement with the accreditation requirements
FEATURES OF PENTRA DF NEXUS - METHODS OF MEASUREMENT
Cytochemistry
Impedance
Flow Cytometry
FEATURES OF PENTRA DF NEXUS - BALANCE CONCEPT
Automatic control of the leucocyte count based on 3 independent principles
FEATURES OF PENTRA DF NEXUS - DHSS
focused flow cytometry and sequential
measurement (impedance and absorbance)
FEATURES OF PENTRA DF NEXUS - Automatic Reflex Testing
selective and programmable (hematology parameters, alarms and flags, demography)
FEATURES OF PENTRA DF NEXUS - Rack rotation mixing of samples
smooth and efficient
FEATURES OF PENTRA DF NEXUS - Quality
complete traceability for each run in agreement with the accreditation requirements
________ regulation for quality control in
automated hematology
CLIA ‘88
electrical impedance is first introduced by
Wallace Coulter
In electrical impedance, ________ are poor conductor of electricity
blood cells
2 chambers filled with a conductive buffered
electrolyte solution
Electrical Impedance
Electrical Impedance is separated by a ______
small aperture
DC current between two electrodes
Electrical Impedance
In Electrical Impedance, as a cell passes through the ________, ________ is impeded and a _______ is generated.
aperture ; flow of current ; voltage pulse
In Electrical Impedance, the number of pulses indicate the number of the ________
blood cells
In Electrical Impedance, the amplitude (height) of each pulse is proportional to the _______.
cell volume
In Electrical Impedance, the requisite condition for cell counting by this method is ___________.
high dilution of sample
an electrical impedance method which is a measurement of cell volume
“coulter principle” - direct current
an electrical impedance method which is a measurement of internal cell structure
“coulter principle” - radio frequency
Each cell flows in a single line through a flow cell.
Optical Light Scatter
A laser device focused on the flow cell
Optical Light Scatter
In Optical Light Scatter, as laser light beam strikes a cell, it is scattered in _________
various directions
In Optical Light Scatter, ______ capture the light
Photodetectors
In Optical Light Scatter, __________ to cell size
Forward Scatter Light (FALS)
_____________ corresponds to nuclear complexity & granularity of cytoplasm
Side Scatter Light (SS) (90°)
Used to distinguish between granulocytes,
lymphocytes & monocytes
OPTICAL LIGHT SCATTER
fluidics is also known as
the flow system
In fluidics (flow system), the sample is injected into a stream of _______ within the ________.
sheath fluid ; flow chamber
fluidics are focused into the center of the stream forming a single file by the principle of _______
- only 1 cell or particle can pass through the ______ at a given moment
hydrodynamic focusing ; laser beam
in fluidics, the _______ is always greater than the ________ ensuring a high flow rate, thus allowing more cells to enter the stream at a given moment.
sample pressure ; sheath pressure
in fluidics, the high flow rate is used for ________ and low flow rate used for _________.
immunophenotyping analysis of cells ; DNA analysis
This method allows the quantitative and qualitative analysis of several properties of cell populations from virtually any type of fresh unfixed tissue or body fluid.
FLOW CYTOMETRY
In Flow Cytometry, The properties measured include a ________, ____________, and ___________
particle’s related size, relative granularity or internal
complexity and relative fluorescence intensity
In flow cytometry, the most commonly analyzed materials are:
o blood,
o bone marrow aspirate and
o lymph node suspensions.
principle of flow cytometry:
flow cytometer is composed of three main components:
- The Flow System
- The Optical System
- The Electronic System
(component of the flow cytometry) cells in suspension are brought in single file past
The Flow System (Fluidics)
(component of the flow cytometry) a focused laser which scatter light and emit fluorescence that is filtered and collected
The Optical System (Light Sensing)
(component of the flow cytometry) emitted light is converted to digitized values that are stored in a file for analysis
The Electronic System (Signal Processing)
Flow cytometry can be used for ________ of fine needle aspirates, and is a powerful adjunct to _________.
lymphoma phenotyping ; cytologic diagnosis
Neutropenia may be ______ or _______ in nature
Immune or Nonimmune
_________ may result from granulocyte specific autoantibodies, granulocytespecific alloantibodies, or transfusion-related anti- HLA antibodies.
Immune neutropenia
Flow cytometry can readily identify __________ that are either bound to granulocytes or free in plasma and confirm the origin of __________, possibly eliminating the need for a bone marrow procedure.
anti-neutrophil antibodies ; neutropenia
Basic Uses of Flow Cytometry
- Phenotyping
- Apoptosis and cell death
- Cell cycle, cell divising and DNA synthesis
- Transduction/transfection confirmation
- Cell tracking
- Small particle analysis
- Functional analysis - calcium flux, gene expression, dye efflux, mitochondrial activity
- Marine and microorganism identification
Potential Applications of Flow Cytometry
- Cell activation status
- Cell cycle distribution
- Cell division
- Apoptosis
- Differentiation state
- Cytokine Secretion
- Activation of signalling pathways
- Calcium flux
- Levels of intracellular reactive oxygen species
- Immune reconstitution studies
- Minimal Residual Disease
Flow cytometry is used in _____________
“Immunophenotyping”
a technique used to study the protein expressed by cells such as erythrocytes, leukocytes, and platelets.
Immunophenotyping
one of the example of immunophenotypung is Immunophenotyping for HLA-B27. HLA-B27
is one of the multiple major histocompatibility
complex (MHC) class I-specific antigens. Detection of HLA-B27 (positive) may associate the patient with several other disorders, such as _________, _______, and ___________.
Reiter’s syndrome, psoriatic arthritis and inflammatory bowel disease
Immunophenotyping of leukemias and lymphomas by differentiating their _________. _______ can also be used to identify leukemias that may be resistant to
therapy
antigen profiles ; Flow cytometry
Flow cytometry not only is can detect the presence or absence of antigens but also the ___________ which can also aid in diagnosis.
strength of antigen expression