????? ??????? (Coulter)

1

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• (????????????? ??????)

2
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3
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4
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  (??????????) ?????????
• ???????????? ??????????? ????? (??????????????
  ????????)
• ???????????? ????????????????? ??????? ?????

5
?????? ??????????? ????????

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  ?????????? ??????????? ??????)
• ????????-???????? ?????????

6
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7
(No Transcript)
8
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9
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1. ??????? ?????? ?????

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1. ??????????????? ??????????

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????????
????????
3. ??????????? ???????????? ???????????
4. ??????????? ???????? ???????? ???????????
(???)
9
10
??????????????? ?????????????? ?????? ?????
???????? ???????????? () ????????? ???????????? (??????/??3) ???????? ??????? (???) ????? (??3)
?????????? 46 (4,2 4,8) 106 7,19,2 1,72,4 70 100
????????? 1,2 (4 8) 103 8 22
?????????? 0,3 (2,5 5) 105 2 4 6
11
????????????? ???????
12
1 - ?????????? 2 - ?????????? 3 - ????????? 4
- ??????? 5 - ????????? 6 - ??????? 7 -
????????.
13
?????? ??? ????????????????? ???????????? ??????
?????
14
????? ????????????? ??????? ??????
15
(No Transcript)
16
????? ??????? (Coulter)
17
????????????????? ??????????? ??????
18

• ?????????? ??????????-????????? ?????????????
  ?????? ??? ???????? ?????? ? ??????????????
  ????????????????? ???????????

19
(No Transcript)
20

• ?????????? ????? ?????? V ????? ???? ????????? ??
  ?????????? ?????????
• V ?2 fkDE/ riF,
• ???
• ? ??????? ??????????? ??????? ?????????,
• fk ??????????? ??????????? ??? ????? ?????????
  ????????? ? ???? ??????????? ??????? ????? ????,
• r - ???????? ????????????? ?????? ?????,
• i ??? ????? ????????? (??????????),
• F ???????????, ??????????? ????? ? ????????????
  ???????,
• D ? ????????? ??????????????? ????????
  ??????????.

21
Cell type Size (fL)
Lymphocytes 35 90
Monocytes 90 160
Granulocytes 160 450
?????? ????? ?????? ?????
22
Number of cells versus cell volume from a Coulter
counter. (a) Nucleated RBCs (N), lymphocytes (L),
mononuclear cells (M), and polymorphonuclear
leukocytes (PMN). (b) Leukocyte differential
distribution (WBC), RBC distribution (RBC), and
platelet distribution (PLT).
23
(No Transcript)
24

• ? ???????? ???????????? ???????????????????
  ????????? ?????? ?????????
• ??????? ???????? ????? ? ????????? ??????
• ??????? ????????????????? ???????????
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  ??????
• ????? ????? ?????, ??????????? ??? ???????
• ??????????? ?????????????? ?????? ?????????????
  ?? ????????
• ???????? ??????????? ? ???????????? ?? ?????????
  ? ????????? ?????? ?????.

25

• ?????????? ??????????????????? ?????????
• ????????????? ??????? ??????????? ??????
  ?????????? ? ?????? ???????????, ????????????
  ???????? ????????.
• ???????????? ?? ????????????? ?????????, ????????
  ?????????????? ? ??????????, ????????? ?? ??????
  ????????.
• ??? ???????? ??????????? ???????????????????
  ??????? ?????????? ????? ??????? ??????????? ??
  ??????? ?????????????? ?????????, ??????? ??????
  ???? ??????? ???????? ???????????.

26

• ???????????? ????????? ???????????? ???????
• ????????? ???????? ?-Z ????????? ??? ????????
  ???????.
• ??????????? ??????? ?????????????? ? ???????
  ???????? Probe Cleaner
• ????????????? ?????? ????????? ????? ???????,
  ??????? ?????? ????????????? ?????.
• ??????? ?? ?????????? ??????? c ??????? ????????
  Probe Cleaner
• ??????? ?????????
• ?????? ?????????
• ?????? ?????? ??? ???????-????????? ??????????
  ???????
• ??????????? ??????? ??? ?????????? ??????????
  ????? 5 ????
• ?? ??????????
• ??? ? 3-6 ??????? ? ??????????? ?? ???????? ??
  ?????? - ?????? ????? ??????? ????
• ?????? ???????? ??????? ??? ????????
• ?????? ??????????? ?????? ?????? ??????????? (1
  ??? ? ??? ??? ???? ? ??????????? ?? ????????)

27

• ???? ??? ???????????????? ???????????? ?? ????
  ??????????? ???????????? ????? ?????????
  ?? ?????? ????
• ? ??????? ???? ????????? ???????, ???????????
  ?????? ?? ?????????? ????? ???????????, ??????
  ?? 68, ? ??? ????????????????? ??????????
  ?? ????????????.
• ?? ??????? ?????? ??????? ???????
  16-20-???????????? ???????????, ??? ??????????
  3-DIFF ???????, ????????? ????????????????
  ????????? ?? ??? ????????????.
• ? ???????? ?????? ????????? ??? ?????????? 5-DIFF
  ???????, ????????? ???????????????? ?????????
  ?? 5 ?????????? ? ??????????? ??????????
  ?? 28 ??????????.
• ????????? ????? ??????????? ? ???????
  ????????????????? ?????????????. ????? ??????????
  ??????????, ???????????? ????????????? ? ?????
  ??????? ??????? ?? 40.

28

• ????????????????? ??????? ?????????? ????
  ???????????????? ??????????? ???????
  ?? ?????????????????? ???????????,
• ? ??? ?????????? ???? ???????? ???????????????
  ?? ??????? ? ???????????? ? ???????????
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  ?????????????? ???????????  ? ???? ???????
  ??????? ?? ??? ??? ??????. ??????? ?????? ??????
  ????????? ???????? ?????? ? ???????????????
  ??????, ?????? ?????? ???????????? ????? ????????
  ??????????????? ? ??????? ??????. ???
  ????????????????? ??????? ????? ??????????????
  ??? ???????????, ??? ? ???????? ?????.

29

• ??????????????????
• ??????? ?????? ???? ??????? ??????????
  ?? 60 ???????? ? ???.
• ??????? ???????? ?????? ????? ??????????????????
  ?? 60 ?? 120 ???????? ? ???. ???????? ??????
  ???????? ???????????? ??? ????? ?????????
  ????????????, ??? ? ????????????? ??????????
  ????.

30

• ????? ???????????????? ????????????????
  ??????????? ???????????? ??? ??????? ?? 10 ?? 300
  ??????????? ??????? ?????. ????? ?????? ??????
  ????? ????????? ???????????? ??????? ? ?????????,
  ? ????? ????? ???????? ??????????? ?????, ???
  ???? ??????????? ?????????? ?????????
  ????????????. ????? ????, ????? ?????? ??????
  ???? ??????? ??????????? ?????????.

31

• ?????????? ?????????? ?????????? ???? ???????
  ???????? ????? ?????????? ????. ?????????? ??????
  ?????????, ???????????? ????????????, ???????????
  ?????? ?? ????????????? ? ???????? ????????????.
  ??????????? ??????? ??????? ????? ???????? ???
  ??????????, ?????????????? ??????-?????????????,
  ??? ? ? ?????????? ?????? ??????????????, ???
  ?????? ?? ??????????? ?? ????????????? ????????
  ????????????, ?? ????? ??????????? ????????
  ?? ????????????????? ???????. ???????? ???????
  ???????? ??????? - ??????? ????? ????????
  ?????? ? ??????????, ??????????????
  ??????-?????????????.

32

• ??????? ????????????? ????????????????? ??????
  ?????????????? ?????????? ???????? ??????????
  ? ????????????? ??????????, ? ????? ???????????
  ? ?????. ????????????? ?????? ? ??????????
  ??????????? ???????? ??????????? ???????????
  ???????. ??????? ? ???????? ???????????
  ???????????, ??????????? ???????? ??????????
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  ??????? ?????????, ???????? ? ????????? ?????
  ???????????? ???????????. ????? ?????? ????????
  ?????????? ??????????? ???????????? ? ??????
  ???????.

33
(No Transcript)
34
(No Transcript)
35

• ??? ?????? ????????????????? ??????????? ???????
  ????????? ????? ??? ????????
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• ????? ????????????
• ?????????????????? ???????
• ?????????????? ??? ?????????????????? ??????????
  ????
• ????? ?????
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• ??????? ????????? ???????? ????????
• ?????????? ????????? ????????

36
????????? ??????????
37

• ??????? ?????? ??????????? ???????? ??????

? ?????? ??????????? ???? ??????? ??????????
?????????? ? ????????? ?????, ??????? ???????????
????????? ????????, ??????, ??????????,
???????????? ? ?????????????? ???????????????
????????
38
(No Transcript)
39

• ??? ??????
• ????????????????? ??? ???????????????????
  ???????????
• ?????????? ???????? ?????? ??????????? ?
  ??????????????? ?????????
• ?????????? ?????? ????????-????????? ?? ??????
  ???????????????

40
?????????? ????? ??????????, ??????????? ??
?????????????? ????????. 1 ???????? ?????, 2
????? ??????????, 3 ????????? ?????? (????? ?
???????? ???? ??????????????? ????????? ?????), 4
??????????????? ?????, 5 ???????? ??? ?????
???????????? ????? ?????. 6 ? 7
???????????????? ?????????? ?????????, 8
???????, 9 ????????? ??????? ? ??????????????
????????????, 10 ???????????? ?????, 11
??????? ?????????, 12, 13, 14 ???, 15 ??????
?? ?????????????? ??????? ??????, 16 ????????
??????????? ?????????????? ??? ?????? ??????, 17
??????????????????? ?????????? ?????????
?????, 18 ?????????????? ???????? ??
??????????? ????? ???????????
41
Optical Detection Scattering

• Theory and Design
• Forward angle light scattering (FALS) passing
  cell scatters the light in the forward direction
  at low angles (0.5 - 10?)
• Photodiodes with filters aligned with laser beam
• Measures cell size
• Orthogonal light scattering light is reflected
  and refracted by subcellular structures
• Coupled with Fluorescence Photomultiplier Tubes
  (PMTs)
• Measures granularity of cell.

Figure from 1
Figure from 3
42

• ???????, ?????????? ??? ????????? ???????
  "?????????? ????", ? - ????????? ??????, ? -
  ??????? ???????, ? - ?????????? ????????? ?
  ?????? ?????????

43
Cell Sorting Droplet

• Theory
• Piezoelectric transducer used to generate
  periodic vibrations.
• Fluid stream is vibrated to form drops that are
  uniformly separated.
• Depending on its characteristics, each drop is
  charged by a strong electrical pulse.
• External electrical field deflects desired cells
  into collecting reservoir.

Figure from 5
44
??????????? ????????? ????? ????????????. 1 -
?????-???????? 2 - ?????????? ?????? ???
??????????? ?????? 3 - ????? ?????? 4 -
?????????? ???? 5 - ???????? ??????????????
???????
45
An automatic analyzer aspirates whole blood,
divides it, dilutes it, mixes it and then
analyzes it for hemoglobin and cell
characteristics. .
46
????????? ????? ?????? ??? ??????????? ?????
???????? ????????
47
(No Transcript)
48
(No Transcript)
49
???????? ????????
50
(No Transcript)
51
?????? ?????????? ??????
????????? ????????? ???????? ?? ??????? ?????
??????????? ??????? ?????
52
. ?????????? ?????? ?????????? ??????
53
??????????? ???????????
54
(No Transcript)
55
??? ???????? ?????????? ?? ??????, ????????? ?
??????????? ??????????? ??????????
56
?????????????? ?????????
?????????
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?????????
??????????
?????????
57
????????? ?????????? ? ????????????????? ???????
???????????
??? ??????????? ????????? ????????????? ??? ?????????????? ??????? ???????????
????? ??????? ????????????? ???????? ??????? (100 000 ? ?????) - ????????? (?????? ?? 1000 ????. 100 000)
???????? ????????? ??????? (100 000/???) - ?????? (?????? ?? 500/??? ????. 5000/???)
?????? ?????????? ????????? - ??????? (??????? ??????????, ?????) ??????? (??????? ??????????)
??????????? ????? ???????????? ??? ??????? - ??????? (?????? ?????? 1 ??, ?????? 100 000 ????????) ?????????, ????? ???? ??????? ????????
??????????? ??????????? ???????? ? ?????? ????????????? - ??????????? ??? ????????????? ?????? 1/10 000 ???????? ??????????? ????????? ????????
???????????? ?????????? ????????? ???????????? - ?????? ????????? ???????
??????? ??????????? ???????? - ??? ????
???????? ??????????????? ?????????? ??????????? ????????? ???????????????? ???????????
58
???????????????
59
??? ???????? ??????????????

• 1. ????????? ??????????? ???????????? ?????
  ?????????? ??????? ?????????? ?????????,
  ????????? ? ?????? ????????? ???????????. ??????
  ???????? ? ???? ???????? ?????????
• 2. ??????? ?????? ????? ? ?????? ? ??????? ??
  5-20 ??????.
• 3. ?????????????????? ????????? ?????? ????? ?
  ??????????? ???????
• ????????????? ?? ???? ?????????? ?
• ?????????? ????????? ? ?????
• ?????????? ????????????? ? ????? ?????????
• ??????????? ??????, ??????? ???????
• ????????? ??????? ??? ???????????? ????????.

60

• ??????? ??????????
• ?????????? ?????????? ?? ?????????
• ?????????? ? ???????????? ? ? ???????
  ??????????? ???????
• ???????????? ??????????
• ???????? ?? ????? ??????????? ????
• ???????? ??? ?????????? ???????
• ??????????? ??????? ?????
• ??????????? ??????? ? ????????????? ?????????
• ??????????????? ??????? ???????? ?????? ?
  ?????????? ??????
• ???????????? ??????? ?????? ? ????

61
??????????????????
?????? ?????????? ????????? ???? ???????????
??????????? ????????? ????? ??????????
???
62
???????????? ??????????? SpO2 ???
??????????????????
Blood Cuvette
Red Light Source 660 nm
Photodetector
Infrared Light Source 810 nm
D(660), D(810) ???????? ?????????? ?????????
????? ?????????? ? ?????????????? ????? ? ? 660
? 810 ?? ? ? ? ??????? ?????????.
63
???????????????????
????????? ???????? ?????????? ??? ???????????
????? ????? ???????? ? ???????????? ????????
64
??????? ???????????
?????????? ??????????? ?????
Ip(lr), Ip(lir) ? Ib(lr), Ib(lir) ????????? ?
?????????? ???????????? ?????????? ?? ???????? ?
????????????? ??????
65
????????????? ?????? ??????????????
66
??????????? ????? ??????????????
67
????????????? ???? ???????????
68
(No Transcript)
69
(No Transcript)
70
??????????? ??????????????? ??? ?? ???????
?????????? ??????????????? ???? ??????? ??????
???????????, ?? ?? ???? ?????? ?????????? ?
??????????????? ?????????? ???????. ???????????
???????. ? ??????????? ??????? ?????????????
?????? ????, ??? ??? ???????? ?????? ? ??? ??????
? ????????????? ?? ????? ?????????? ????????????
??????. ??????? ????????? ?????. ???? ???
??????? ????????? ????? ?? ??????????????, ?????
????? ???????? ????????? ???????????. ??????????
????? ??????? ????, ?????, ???????? ?????
????????? ????????????? ?????? ? ????????
????????? ??? ??????. ??????????? ????
??????????? (????????, ????????????? ???
????????????? ??????????) ????? ?????? ????????
????????? ?? ?????? 85. ??????????????????,
???????????? ??? ?????????? ??????? ?????, ?????
?????? ???????? ????????? ????? 100. ?????????,
??????? ??? ??? ??????, ????? ??????????????
?????????? ???????? ?????????. ??????????????? ?
?????????? ???????? ?????????? ???????? ?????? ?
???????? ??????????? ???????. ???????? ?????????
???? 70 ?? ??????, ?.?. ??? ??????????? ????????
??? ?????????. ????????? ????? ?????? ?????
???????? ?????????? ??????????????? ??????????
???????. ????????????? ???????. ??? ??????, ???
??????????? ???????? ????????? ? ????? ?????
????????? ??????? ???????, ??? ?????? ?????????
?????????. ???????? ??????? ??????? ? ???, ???
?????? ???????????. ??? ??????, ??? ??????????
???????? 5-20 ?????? ????? ???, ??? ????????
??????????? ????????? ???????? ?????? ?
?????????? ???????? ?? ??????? ??????????????.
71
Tissue spectrometer
72
Molecule Type of test Normal levels US units (mg/dL) Normal levels SI units (mmol/L) Toxic levels (mg/dL)
Total bilirubin Blood (serum) 0.21.0 3.417.1?103 High levels result in jauntice
Lactate Blood (serum) 512 0.51.3 gt45
Creatinine Female Male Blood (serum) 0.61.1 0.71.2 5397 ? 103 62115 ? 103 gt4
Urea Blood (serum) 1020 3.67.1 gt100
Glucose Blood 70105 3.95.8 lt50 or gt400
Sodium Blood (serum) 136145 mEq/L 136145 lt120 or gt160 mEq/L
Potassium Blood (serum) 3.55.1 lt3 or gt7.5 mmol/L
Lithium Blood (serum) 0.81.2 mEq/L NA gt2.0 mEq/L
Table 3.1 Normal and toxic levels of various
molecules in the body given in both US and
European (SI) units (Pagana and Pagana, 1995). NA
denotes not available.
73
????????????? ?????????? ??????? ????????????

• ????????????? ?? ???????????? ???????????????
  ??????????? ?????????
• ?) ???????????????
• ?) ??????????????????????
• 2. ????????????? ?? ???? ?????????????? ????????
  ? ??????????
• ?) ????????????? ??????????
• ?) ????????????
• ?) ?????????????
• ?) ??????????????
• ?) ??????????? ??????????
• ?) ?????????????? ??????????
• 3. ????????????? ??????? ?? ????????
  ????????????
• ?) ?????? ???????????? ???????? ? ????????
  (?????????????)
• ?) ??????, ??????????????? ??? ????????????
  ?????????.

74
?????????? ????????????? ???????

• ????????? ? ????????????????
• ????????????
• ??????????? ? ??????????????????
• ????????? ?????????
• ???????????
• ???????????

75
??????? ??????

• ??????????????? ?????? ???????????? ?????????? ??
  ??????????? ?????? ???? (?????????) ?????????
  ???????? ?????????.
• ? ?????? ?????????????? ?????? ??????? ???????
  ?????????? ????? ??????????????????. ??
  ?????????? ?? ???? ???????.
• ????????????? ?????????? ??????? ?????????
  ??????, ???????????? ??????, ?? ??????? ??
  ????????????? ?????????. ?????? ??????????? ????
  ?????????? ??????? ????????? ?????? ????
  ??????????? ????? ??? ???? ??????????????????
  ????????? ??????.
• ?????????? ?????????????????? ?????? ????????
  ??????? ????? ??????????????? ????? ???????
  ???????????? ????????.
• ???????? ??????????? ? ?????? ?????????? ?
  ????????? ? ???????? ? ????????? ?? 0 ?? 100.
  ?????????? (?????????) ?, ?????????? (??????????
  ?????????) ? ???????? ????????????. ????? ???
  ??????????? ? ?????? ??? ???????? ??????????
  ?????????.

76
?????????? ?????
???
???
77
????? ?????????????? ??????????????
???????????????

• 1 ???????? ????????? 2 ??????????
  ????????????? ???????, 3? ??????????? ????????,
  3? ???????? ?????????, 4 ????????????
  ??????????, 5 ?????????? ??????????????
  ??????????, 6  ?????????? ??????????? ?
  ??????????? ??????????.

78
(No Transcript)
79
????????? ??????????? ????????????? ?????????
80
????? ????????????? ?????????????? ?????????
81
???????????????
??????????????? ??????????? ??? ??????? ?????????
?????? ??????????. ????????? ?????????? ?
????????????? ????????? ? ??????? ???? ???? ????
461 ?? ? 551 ??. ???????? ?????????? (???????
????????????????) ??? ?????????? ? ?????????
??????????? ??? 461 ?? (??? ???????? ??
????????). ?? ? ??????????? ???????? ?????
????????? ? ?????? ??????, ????????, ??????????
?????????? ????????, ???????, ??? ???????
???????? ? ?????? ??????? ???????????? ??????
?????? 551 ??. ???????? ????????? ?????????? ?
???? ??????? ?????????? ??????????, ?????????? ??
???? ????????. ???????????? ?????? ?????
??????? ???????? ????????? ?????? ??????????
0513 ??????/?, ??????????? ??????????? (5),
????? ????? ????? ????? (2 ?????), ???????
?????????, ??? ????????????? ? ?????????.
82
????????????????

• ???????? ??????? ???????????????? ??
  ?????????????????????? ??????? ? ???????????
  ?????????? ????? ??????????? ????? ???????? ?????
  ????? ?????? ????? ?????, ?????????
  ??????????????? ?????????, ???????? ???? ????????
  ???? ???? ?? ?????? ???????? (VIS) ????? ?? 380
  ?? 750 ??, ?? ? ???????? ????????????? (UV) ??
  190 ?? 380 ??.

83
?????????? ??????????? ????? ??????????????
????????????????
84
?????????????

• ?????? (220 950 ??)
• ????????????? ??????? (200 800 ??)

85
????????? ?????????

• ?????????? ??? ??????????? ????????????? ?????
  (200 - 360 ??)
• ??????????? ????? (200 - 360 ??)
• ????? ??????????? ? ???????????? ????? (360 800
  ??)

86
????????????? ???????????
????????????? ??????????? ????????????? ???
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87
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88
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???? ????? ??????????????? ??????????? ABBOTT
Spectrum
91
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?????
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The flame photometer aspirates a sample
containing metal ions and heats it to
incandescence. Detector output is proportional to
concentration
95
Figure 3.3 LifeScan, Inc., a system by Johnson
and Johnson for self-monitoring glucose levels.
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Figure 3.5 In the glucose enzyme electrode, when
glucose is present, it combines with O2, thus
decreasing the O2 that reaches the cathode.
98
Figure 3.4 In the PO2 electrode, O2 dissolved in
the blood diffuses through a permeable membrane.
Current is proportional to PO2 (Hicks et al.,
1987).
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Figure 3.19 In an electrophoresis system, charged
molecules move through a support medium because
of forces exerted by an electric field.
101
v velocity
E strength of the electric field
q net charge of the molecule
f molecules resistance
Migration distance
Figure 3.20 This serum protein electrophoresis
demonstrates a normal pattern, with the largest
peak for albumin.
102
Migration distance
Figure 3.21 This serum protein electrophoresis
demonstrates a decrease in the albumin and an
increase in gamma globulins.
103
???????????? ??? ????????????????? ???????
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2100
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????????? - ?????? StatFax 2200
????????????? Awareness Tehnology, ???

104
ELISA microplate reader



105
Hidex Chameleon microplate reader

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????????? ???????? ?????
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?????????????????? ?????
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115

• ???. 1. ????-????? ???????? ?????????????
  ???????? (????? ?????? ???????? ??????,
  ??????????? ?? ??????????? Freescale
  Semiconductor)

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Plethysmography (3)Electric-Impedance Method
Different tissues in a body have a different
resistivity. Blood is one of the best conductors
in a body ( 1,5 Om)
A constant current is applied via skin electrodes
I 0,5 4 mA rms (SNR) f 50 100 kHz
(Zskin-electrodeshock)
The change in the impedance is measured
The accuracy is often poor or unknown
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  ????????? ?????? ???????, ? ??????????
  ??????????????? ?????? (????????????).

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Thorax impedance curve
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Impedance Tomography
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