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HEMA LEC 1 > TOPIC 3: AUTOMATION IN HEMATOLOGY > Flashcards

TOPIC 3: AUTOMATION IN HEMATOLOGY Flashcards

1
Q

_______started with the ingenuity of the Coulters.

A

Automation

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2
Q

Used in order to count and size the different blood
cell

A

COULTER PRINCIPLE

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3
Q

Why do we employ analyzers in Hematology?

A

● Cell counting
● Diagnosis of hemoglobinopathies
● Immunophenotyping
● Diagnosis of Leukemias and Lymphomas
● Coagulation abnormalities

CDIDC

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4
Q

Complement system
- Anemia caused by a deficiency in _______ and ________
- With the use of hematology analyzing machines,
physicians may be able to directly diagnose based on
the results from a particular laboratory analyzer

A

Paroxysmal nocturnal hemoglobinuria
CD55 and CD59

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5
Q

ADVANTAGES OF AUTOMATION

A
  1. Speed and efficient handling of samples
  2. Greater Accuracy and Precision
  3. Multiple tests on single platform
  4. More efficient workload and management
  5. More timely diagnosis
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6
Q

DISADVANTAGES OF AUTOMATION

A
  1. Flagging of machine
  2. RBC morphology limited
  3. Erroneous results
  4. Expensive
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7
Q

● Measures only few parameters
● Some steps like dilution is carried out manually
○ Diluted sample will be fed to the machine

A
  1. SEMI-AUTOMATED
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8
Q

● All of the steps during analysis are performed by the
machine itself
● what MTs do is load the sample on the machine and
press button - analysis will start
● Measures multiple parameters
● Requires only anticoagulated blood samples

A
  1. FULLY AUTOMATED
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9
Q

● Vacuums and pressure for operating valves and also
to move the samples among the hydraulics

A

Pneumatics

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10
Q

● Analyzers and computing circuitry
○ In order to process the data during analysis
● Responsible to do the analysis and then the
calculation, and also the reflection of the sample of
the result generated
● Will be able to control the operational sequences of
the total system
● has an algorithm on how to analyze and process the
data during analysis of the blood samples/ blood cells

A

Electricals

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11
Q

Makes use of the Coulter Principle
○ Detection and measurement of changes in
electrical resistance produced by cells as
they traverse a small aperture

A

Electrical Impedance

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12
Q

Where the sample is suspended
*solution is composed of electrically conductive diluent
● There is a flow of electric current within the solution
● 2 chambers filled with a conductive buffered
electrolyte solutions separated by glass tube having
a small aperture
● direct current is generated between the internal and
external electrode
● Aperture for RBC/platelet is smaller than the WBC
aperture

A

APERTURE BATH

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13
Q

has the aperture where the cells pass through

A

Aperture tube

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14
Q

➔detects/does the analysis of the
blood cell that passes through here
● Internal And external electrode

A

Sensing zone

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15
Q

BASIC COMPONENTS OF HEMATOLOGY ANALYZER
Hydraulics

A

○ Mixing Chambers
○ Aperture baths/Flow cell
■ Where the samples are suspended
for anlaysis
○ Hemoglobinometer
■ Where Hgb is measured
○ Aspirating unit
○ Dispensers
○ Diluters

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16
Q

The height of the pulse is equivalent to the volume of
the cell
● Higher pulse, higher volume
● Number of pulses is equivalent to the number of cells
analyzed in the machine
● The pulses will be gathered and sorted out so they
will be plotted into the histogram

A

Oscilloscope

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17
Q

volume of the cell

A

X axis

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18
Q

number of the cell

A

Y axis

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19
Q

➔Through this analysis, the cells will be able to be counted
and will be discriminated/differentiated
➔Basis of differentiation will be through their CELL VOLUME

A

THRESHOLD DISCRIMINATOR

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20
Q

The machine is set to identify a ______________
on a treshold volume

A

particular cell based

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21
Q

Provides a sample stream surrounded by a sheath
fluid as cells pass through the aperture
● Allows the alignment of cells into a single-file
passage through the sensing zone

A

Hydrodynamic Focusing

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22
Q

● Incorporated with another principle
There is application of high voltage electromagnetic current
flowing between electrodes

A

RADIO FREQUENCY

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23
Q

● Involves light scattering

A

OPTICAL LIGHT SCATTER

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24
Q

➔Light Amplification by Stimulated Emission
of Radiation

A

LASER

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25
Q

There is a source of light → in the form of _______________

A

TUNGSTEN HALOGEN LAMP / HALOGEN NEON
LASER

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26
Q

The blood cell will scatter the light in
different directions ; hence the name _______________

A

Light scattering

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27
Q

The light scattered by the cell will be captured
___________, ________, ___________, _____________

A

● Absorption, diffraction, refraction, and reflection

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28
Q

There is a single file of cell; because this principle also
employs

A

HYDRODYNAMIC FOCUSING

29
Q

Different angles of light scattered detected by
detectors

1.a. 0 degree scatter – straight line
b. Diffraction of light
c. Cell volume

2.a. A.k.a Orthogonal Light Scatter
b. Refraction and reflection of light
c. Internal structures of the cell

3.a. → Differential scatter
b. Cell volume

A
  1. Forward angle scatter
  2. Side Scatter Light
  3. Forward Low Angel Scatter/Forward High Angle
    Scatter
30
Q

_______________ - directly count RBC

● ________________ – direct measurement
o It is able to directly measure the cell
through forward angle scatter

● __________________
o It may be 3-part or 5-part differential; but
with optical light scatter; it is 5-part

A

RBC Count
Mean Cell Volume
5-part WBC differential

31
Q

– light emission when electrons are
raised to an excited state; as they go back to their
ground state; they emit a light of a particular
wavelength

A

Fluorescence

32
Q

FLUORESCENT FLOW CYTOMETRY

1.
2.

A

Light scattering
Fluorescence

33
Q

_______________ - internal cell structure Side

● ____________ - RNA/DNA information
Forward

●_____________- cell volume

A
  1. Side scatter light
  2. Fluorescence light
  3. Scatter light
34
Q

o The sample is injected into a stream of
sheath fluid within the flow chamber
o The cells are hydrodynamically focused so
that they will line in a single file; so these
cells will be analyzed one cell at a time

A

1 - FLUIDICS (THE FLOW SYSTEM)

35
Q

Two types of flow rate

_____________________= immunophenotyping
analysis
______________________ = DNA analysis

A

High flow rate
Low flow rate

36
Q

__________light is required to excite the cells
o Argum/neon helium lamp

A

Laser

37
Q

____________ is proportional to the
amount of light striking a cell

A

ELECTRONICS

38
Q

___________ is collected and stored in the computer

A

Data

39
Q

Data plots

o ____________ = histogram
o ____________ = data plot

A

Single parameter
Two parameters

40
Q

Boundary that can be set to restrict the analysis to a
specific population within the sample

Data selected by the gate is then displayed in
subsequent plots

A

GATING

41
Q

● Consists of collecting cells of interests
o Defined through criteria of size and
fluorescence

A

SORTING

42
Q

Reticulocyte counting makes use of
___________________ to stain the residual RNA
of the reticulocyte

A

supravital stains

43
Q

FLUORESCENT DYES USED FOR SUPRAVITAL STAINING

1.
2.
3.
4.
5.

A

● Auramine O
● Thiazole Orange
o Most widely used fluorochrome for
[residual RNA]
● CD4K 530
● Oxazine 750
● New Methylene Blue

44
Q

Classification of reticulocytes into 3 maturation stages

1.
2.
3.

A

a. Low Fluorescence Reticulocytes - more mature
b. Middle Fluorescence Reticulocytes
c. High Fluorescence Reticulocytes - immature

45
Q

● Identification and counting of granulocytes

A

Peroxidase

46
Q

contain the enzyme myeloperoxidase

A

Granulocytes

47
Q

● Reticulocytes and platelets
● Best for detecting immature
platelets

A

Fluorescence

48
Q

● Accurate platelet counting using
CD41/CD61 antibodies

A

Immunological

49
Q

Makes use of Von Behren’s plate

A

Abbott CELL-DYN

50
Q

With MAPSS technology, the light that is scattered at
90 degrees will be directed to a __________

A

polarizer

51
Q

● Makes use of a unifluidics instead of hydraulics

A

Siemens ADVIA 2120i

52
Q

It employs peroxidase staining for identification of
granulocytes, particularly neutrophils

A

Siemens ADVIA 2120i

53
Q

RBCs, platelets, neutrophils, lymphocytes,
monocytes, eosinophils will be
disintegrated upon exposure to _____________, and basophils will remain intact

A

non-ionic surfactant

54
Q

________, ________, ________make use of Modified
Cyanmethemoglobin which makes use of Drabkin’s
reagent

A

Beckman, Abott, and Siemen

55
Q

For _________, it makes use of Sodium lauryl sulfateHGB technique

A

Sysmex

56
Q

Graphical representation of numerical data of
different cell populations in a cell counter
● Gives information on:
o Average size of cell
o Distribution of size

A

HISTOGRAM

57
Q

● The normal curve peaks between _______

A

80-100

58
Q

________________ will be able to identify the
type of cell based on its volume

A

Threshold discriminator

59
Q

● Platelets have volume between 8-12 fL and counted
between ________
o Some references say 2-30

● RBCs have volume 80-100 fL and are counted
between __________

● Peak of RBC MCV – ________

A

2-25

25- 250 fL

80-100 fL

60
Q

Small cytosis/cells
o Most of the cells analyzed by the machine
have a very small volume- so if we take a
look at this under the microscope, we will
be able to observe cells that are very small
o Cells having a volume of around 50 fL – very
small (normal volume is around 80-100)

A

● MICROCYTOSIS

61
Q

There is a peak in the volume greater than
100 fL

A

MACROCYTOSIS

62
Q

Significant number of cells having smaller
volume

A

● DIMORPHIC POPULATION

63
Q

To ensure readings from an instrument are
consistent with other measurements
- To determine accuracy of the instrument readings
- To establish reliability of the instrument
When the machine is able to give measurements that
are accurate over a long period of time, that indicates
that the machine is reliable – we can trust the results
generated to be___________ and ______________

A

Accurate and precise

64
Q

● The values are known

● Done
o Upon installation of machine
o Every _________
o Periodic after major repair

A

Commercial Calibrators

6 months

65
Q

Lipemic samples will cause __________________

A

FALSELY INCREASED MCHC

66
Q

________________ the sample is milky due to the
high lipid content in the sample of the
patient
o Causes turbidity in the sample
o False increase in the hemoglobin reading
o MCHC will be falsely increased – this is
directly affected by hemoglobin

A

Lipemia

67
Q

_________________the cells are rupturing
o Causes false low increase
o Increase in the hematocrit during
hematologic analysis – Hematocrit is a
computed value(not directly measured)
▪ Equal to MCV x RBC count /10

A

Hemolysis

68
Q

___________________There will be false increase because
multiple cells are analyzed at the same time
o The cells seemingly become one big cell

A

● Cold agglutinins, RBCs appear big

69
Q
A

HEMA LEC 1 (3 decks)

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