TRUNG TM Y T KHNH VNH KHOA XT NGHIM - CHA

NGUYN L HOT NG
CA MY PHN TCH T BO MU T NG

(Lu hnh ni b Khoa)

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u __________

Nguyn l c bn ca my m t bo theo dng (flow cytometry) l nguyn l bin i in tr ca dng ht i qua ca s c t bo quang in v mt in trng. Nguyn l ny gip phn tch s khc bit v kch thc cc loi t bo khc nhau nhng khng nhn din chnh xc tng loi t bo. Cc my m t bo hin nay ang c s dng c th chia lm 2 loi: 1) My m t bo nguyn l tng tr: phn bit tng loi t bo da vo kch thc t bo. 2) Cc my th h sau: ng dng laser v xung in a chiu nn c tc cao v phn loi t bo chnh xc hn. - Vi nhng my sn xut trc nm 1996, kh nng phn loi chnh xc cc thnh phn bch cu ni chung khng qu 90%. - Cc my model gn y, vi vic p dng tng hp cc c ch tng tr, xung in a chiu, laser nn kh nng nhn din t bo c nng n 95%.
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Mt s serial my c th phn bit c cc loi bch cu a acid, a baso, hng cu li bng vic kt hp vi cc phng php nhum men Peroxydase, nhum ARN/AND, nhum hunh quang, phn tch huyt sc t (CellDyn 4000 ca hng ABBOTT, Se-Advance ca hng Sysmex)

Bi vit ny xin cp n nguyn l hot ng ca my Sysmex KX21 v nhng bin lun xung quanh vn ny./.

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NGUYN L HOT NG CA MY HUYT HC SYSMEX KX-21 V BIN LUN CC THNG S KT QU

I. NGUYN L M T BO MU TRN MY SYSMEX KX-21 1. Phng php in tr khng:
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Mu mu c ht v pha long trong dung dch in phn theo 1 t l nht nh. Sau c dn vo bung m.

DC

Cu to bung m: 1 khe m. 2 in cc (m, dng). Dng in 1 chiu i qua 2 in cc.

Phng php in tr khng (+) (-)

C R

V t bo mu c coi nh khng dn in nn khi 1

t bo mu i ngang qua l khe m s lm dch chuyn mt lng dung dch mui tng ng vi th tch t bo; ng thi lm ngt qung dng in, dn n thay i hiu in th gia 2 in cc (tng in tr), to ra 1 xung in.
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Cc xung in thay i ny c th m c, th hin s lng t bo. ln ca mi xung t l vi th tch ca tng loi t bo. Trung bnh ln ca cc xung in chnh l th tch trung bnh ca t bo (MCV).

1.2 Phng php quang hc (o nng Hemoglobin):

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Hng cu c lm ly gii v gii phng Hb. Sau , Hb c chuyn thnh Cyanmethemoglobin, mt quang ca dung dch ny (o bc sng 555 nm) t l thun vi lng Hb trong mu.

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II. S H THNG PHN TCH CA MY SYSMEX KX-21

(1.960 mL) (1.996 mL) Bung pha long (4 L)

Dung dch pha long (1:500) (1.994 mL) (1.0 mL) H thng ht mu (6 L) Mu ton phn (50 L)

Bung m HC/TC (40 L) (1:25000) (1:500) Bung m Bch cu

Dung dch ly gii

Bung o Hb (1:500)

1. Nguyn tc chung trong m s lng t bo:

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T bo c kch thc 02 030 fL: Tiu cu. T bo c kch thc 25 250 fL: Hng cu.
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T bo c kch thc 30 300 fL: Bch cu.

+ T bo c kch thc 030 080 fL: Lymphocytes. + T bo c kch thc 080 130 fL: Monocytes, Basophils, Eosinophils.

+ T bo c kch thc 130 300 fL: Neutrophils. 2. Phn tch Bch cu/Hb:
Dung dch pha long (1.994 mL) Dung dch ly gii (1 mL) Bung m Bch cu (1:500) Mu ton phn (6 L) 4 Bung o Hb (1:500)

H thng ht mu

2.1 Ti bung m Bch cu:

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Mu sau khi c pha long s b ly gii thnh phn Hng cu. Thnh phn cn li: Bch cu, Tiu cu, Hng cu c nhn, Hng cu c KSTSR, Hng cu c bn cao (trong bnh Thalassemi).

Bin lun:
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Tiu cu c th tch trung bnh 10 fL (2 30 fL). Nu 4 Tiu cu dnh li vi Gi s SLTC bnh thng l 200 G/L, SLBC bnh thng 10 G/L. 200.109 Tiu cu tng ng vi 20.109 Bch cu.

nhau s c kch thc 40 fL, tng ng vi 1 Lymphocyte. + Nu SLTC trn ngng tp theo nguyn tc 10 Tiu cu = 1 Bch cu th

+ Nh vy, SLBC tng gi to thnh 10 + 20 = 30 G/L.

Kt lun:
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Nu Tiu cu gim gi to (do ngng tp, kt dnh) s lm tng SLBC. Thc t, nu SLTC gim, SLBC tng:

+ Cn phi ko lam nh gi ngng tp Tiu cu. + m li SLBC trn bung m. p dng thc t: trnh s ngng tp, kt dnh Tiu cu do ch quan, cn phi: Lng cht chng ng tng ng vi th tch mu. Sau khi ly mu xong phi cho ngay vo tube chng ng v lc trn u k

ngay sau .
2.2 Ti bung o Hb:

Nguyn tc: Hng cu c lm ly gii v gii phng Hb. Sau , Hb c chuyn thnh Cyanmethemoglobin, mt quang ca dung dch ny (o bc sng 555 nm) t l thun vi lng Hb trong mu. Bin lun:
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Hb c o theo nguyn tc quang hc. Do , nhng nguyn nhn no lm

tng mt quang ca huyt tng s lm tng nng Hb gi to. Ngc li,

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i vi nhng trng hp gim nng Hb gi to c th do Hng cu cha c ly gii hon ton.

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Nhng nguyn nhn lm tng nng Hb gi to:

+ Tng Lipid mu (huyt tng c mu trng). + S lng Bch cu tng cao, trn 50 G/L (huyt tng c mu trng).

+ Vng da (huyt tng c mu vng). + Tan mu (huyt tng c mu ). Nhng nguyn nhn lm gim nng Hb gi to: + Dung dch ly gii hng (Hng cu cha ly gii hon ton).
+ Hng cu b ngng kt (ch ly gii 1 phn ngoi ca cm Hng cu).

p dng thc t: -

Tt nht nn ly mu lc bnh nhn cha n ung g (trnh tng Lipid mu). Trnh ly gii Hng cu trong qu trnh ly mu. Lng cht chng ng ph hp vi th tch mu. Sau khi ly mu xong phi cho ngay vo tube chng ng v lc trn u k

ngay sau . 3. Phn tch Hng cu/Tiu cu:

Dung dch pha long (1.960 mL) (1.996 mL) H thng ht mu Mu ton phn (4 L) (1:500) Bung pha long (40 L) Bung m HC/TC (1:500)

Thnh phn trong bung m Hng cu: Hng cu, Tiu cu, Bch cu. Nh vy, da vo nguyn tc phn bit t bo theo kch thc th Bch cu (c khi Tiu cu khng l) s c m vo trong Hng cu.

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 Bin lun 1:
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Hng cu tng gi to do Bch cu tng qu cao ( > 50 100 G/L). Tiu cu tng gi to:
+ Do Hng cu v thnh nhiu mnh.

+ Do Hng cu kch thc qu nh (MCV < 65 fL). + Do trong mu mu c bi. + Do my tip t khng tt gy hin tng nhiu nn cc xung in nh s c my ghi nhn l cc Tiu cu.
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Tiu cu gim gi: mu vo tube thy tinh l Tiu cu kt dnh vo thnh tube ri hot ha v kt dnh.

+ Do Tiu cu b ngng tp: ly mu qu lu lm hot ha Tiu cu hoc ly

+ Do Tiu cu tp trung xung quanh Bch cu. Bin lun 2: nh gi Ngng kt Hng cu Ngng kt Hng cu c th do: + Ngng kt nng: xy ra nhau trong c th (do Khng th). + Ngng kt lnh: xy ra khi mu ra khi c th. Khi o, kt qu phn tch trn my s l: + Bch cu: + Hng cu: + Tiu cu: + Hb: + MCV: + HCT: + MCH: + MCHC: Tng/Bnh thng. Gim nhiu. Bnh thng (khng nh hng). Bnh thng/Gim nh. Tng c mc (<125 fL). Gim nhiu (do Hng cu gim nhiu). Tng nhiu. Tng nhiu.

Kt lun: nh gi s Ngng kt Hng cu, ta da vo: MCH v MCHC (tng nhiu). SLHC v HCT (gim nhiu).
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 Tm tc: Cc nguyn nhn gy sai s v s lng Hng cu

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Hng cu b ngng kt (tan mu t min): s lng Hng cu gim, MCV

tng, MCHC cao (trn 380 g/L). Trng hp ny s dng c kt qu Hemoglobin, cn Hematocrit th s dng phng php ly tm vi th tch hoc mu mu 370C trong vng 30 pht ri m li. Cn kim tra k ng mu khi nhn bnh phm. mu. Lc ng mu khng k: nu ht phn trn ca ng mu s gy hin tng gim Hng cu r rt v khng tng ng vi tnh trng lm sng. Nu ht phn y ng s gy tng Hng cu gi to, ng thi gy gim r rt s lng Tiu cu. Cn kim tra v i chiu s lng Tiu cu trn lam nhum Giemsa. Do Hng cu nh, Tiu cu to, cm Tiu cu, mnh Hng cu, tan mu, ri Mu b ng, Hng cu b v hoc lng mu ly lm xt nghim khng Ch : MCHC (g/L) 320 360 < 300 300 360 > 360 ngha Bnh thng Hng cu nhc sc Hng cu bnh sc Khng khng nh c Hng cu u sc (V Hng cu bo ha Hb, khng dung np thm) lon ng mu, Bch cu qu nhiu u dn n sai kt qu. Tng nht huyt tng: vi thi gian v p lc ht thng thng ca my m t bo c th gy ra gim 3 dng ngoi vi gi to do mu b qunh nn my ht khng

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III. CNG THC TNH TON V SUY LUN 1. Cng thc tnh ton cc thng s:
HCT RBC HGB MCH (Mean Corpuscular Hb): MCH = RBC

MCV (Mean Corpuscular Volume): MCV =

MCHC (Mean Corpuscular Hb Concentration): MCHC =

HGB HCT

2. Cc thng s my Sysmex KX21 o c:

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MCV: Trung bnh ln ca cc xung in o c. S lng Hng cu: Tng s xung in o c ti bung m Hng cu/Tiu cu. S lng Bch cu: Tng s xung in o c ti bung m Bch cu/Hb. S lng Tiu cu: Tng s xung in o c ti bung m Hng cu/Tiu cu. Lng Hemoglobin: T l thun vi mt quang ca dung dch mu sau khi lm ly gii Hng cu v o bc sng 555 nm. S lng Neutrophils: Tng s xung in o c ti bung m Bch cu/Hb. S lng Lymphocytes: Tng s xung in o c ti bung m Bch cu/Hb. p dng cng thc tnh ton v cc thng s my o c, t suy lun ra cc thng

3. Suy lun: s khc nh MCH, MCHC, NEUT%, LYM%...

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IV. BNG GI TR CC THNG S HUYT HC BNH THNG

STT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 WBC RBC HGB HCT MCV MCH MCHC PLT LYM% LYM# NEUT% NEUT# MXD% MXD# MO% MO# BASO% BASO# EO% EO# RDW PDW MPV P-LCR RDWSD RDWCV K hiu White Blood Cells Red Blood Cells Hemoglobin Hematocrit Mean Corpuscular Volume Mean Corpuscular Hb Mean Corpuscular Hb Concentration Platelet LYM% LYM# NEUT% NEUT# MID% MID# Monocytes% Monocytes# Basophils% Basophils # Eosinophils% Eosinophils# Red Distribution Width Platelet Distribution Width Mean Platelet Volume Large Platelet Ratio ngha S lng Bch cu. S lng Hng cu. Hemoglobin. Th tch khi Hng cu. Th tch trung bnh H.cu. Lng Hb trung bnh H.cu. Lng Hb trung bnh 1 H.cu. S lng Tiu cu. % Lymphocyte. S lng Lymphocyte % Neutrophil S lng Neutrophil. % Monocytes, Basophils, Eosinophils. S lng Monocytes, Basophils, Eosinophils. % Monocytes S lng Monocytes % Basophils S lng Basophils % Eosinophils S lng Eosinophils phn b Hng cu. phn b Tiu cu. Th tch trung bnh Tiu cu T l tiu cu khng l. Di phn b HC- lch chun Di phn b HC-H s bin thin Gi tr Bnh thng 4.0 10.0 G/L Nam: 4.2 5.4 T/L N: 4.0 4.9 T/L Nam: 130 160 g/L N: 120 142 g/L Nam: 0.40 0.47 L/L N: 0.37 0.42 L/L 85 95 fL 28 32 pg 320 360 g/L 150 500 G/L 25 40 % 1.2 4.0 G/L 55 70 % 2.8 6.5 G/L 5.1 13.2 % 0.22 1.32 G/L 14% 0.05 0.40 G/L 0.1 1.2 % 0.01 0.12 G/L 48% 0.16 0.8 G/L 11 14 % 11 15 % 5 8 fL Gi tr thc t o c 35 46 fL 11 14.4 %

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