Flow-cytometrie
Měřící princip
Měření rozptylu světla na buňkách a biologických částicích (nefelometrie)
Měření fluorescence obarvené DNA a RNA
Měření impedance procházejících buněk a biologických částic
(elektrochemická metoda)
Měření absorbance hemoglobinu (spektrofotometrie)
Merici zanzeni
Optický systém
Průtoková optická cela (leukocyty, erytrocyty, trombocyty, retikulocyty)
Komůrka k měření impedance (el. odporu) erytrocyty, trombocyty
Průtoková spektrofotometricka cela (hemoglobin)
Optical Bench
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Multi Angle Polarised Scatter Separation (MAPSS)
0° Light Loss:     SIZE
7° Scatter:          COMPLEXITY
90° Scatter:        LOBULARITY
90° D Scatter:    GRANULARITY
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Ou Light Loss
</   0° Light Loss is
proportional to cell SIZE
S   Full light intensity = no cells in the laser beam
^   Decreased light intensity = cells in the laser beam
^   Detected by the inner element of the Bull's Eye Detector
No Cell Present
Cell Present
ft
B
DeCOCttr Qylpul
ÍTĽylil
Tmfc
Detector Output
L. "*"
O   Litjľ.l
Stg-ai
Tima
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7° Light Scatter
S A cell with a complex internal structure produces a larger 7° Light scatter signal than a cell of low complexity
S 7° Light Scatter can be used to measure cellular complexity
S Greater cell complexity = greater low angle scatter
S  Detected by the outer element of the Bull's Eye Detector
a
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90 Light Scatter
S 90° Light Scatter is used as a measurement of the lobularity of a cell's nucleus
S 90° Light Scatter is a function of NUCLEAR LOBULARITY
S Greater lobularity = greater 90° Light Scatter
S 90° Light Scatter is detected by a photomultiplier tube (PMT2)
PSS                                                     PSS
EJ
Argon ion          III                            Argon ion
laser light                      ^1Ab          laser light          _         ^ IAS
- ^*W   — ALL                        -
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ALL
90  Depolarized Light Scatter
S 90° Depolarized Light Scatter can be used to differentiate eosinophils from neutrophils
S Granules found in eosinophils, depolarize light during the course of scattering it
S 90° D is detected by a photomulitiplier tube (PMT1)
DSS    PSS                                                    DSS    PSS
i
Argon laser      étĚt^                                 ArSon laser
light source                       J>IAS                   Ught source
ALL
ALL
a
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■

Fluorescence
Red fluorescence is a measurement of the amount of
DNA stained by the dye.
Axial light loss (0°) measures cell size.
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WBC Differential	
S Differential Analysis by MAP PS	
S Cell Populations are colour coded	^H    m  -j
S Flags to alert of abnormalities	
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