細胞健康檢測指標：細胞活性（Cell Viability）
 
由於死細胞對於抗體有較強的非特異性結合，所以當進行流式細胞儀實驗時，在樣本準備上利用cell viability dye進行染色，透過螢光表現差異圈出並排除死細胞，能讓分析結果的統計更加精確。
 
挑選cell viability dye時需考慮以下事項：
1. 是否搭配需內染的細胞實驗：常見的染劑對於固定液和打洞液不具耐受性，只適用於做細胞表面染色的實驗，需內染的實驗則要選用對固定液和打洞液具耐受性的商品。
2. 是否適合流式細胞儀上機分析：先確認所用流式細胞儀設備規格，挑選可經雷射正常激發並偵測到螢光訊號的染劑。
 
嘉義長庚貴重儀器實驗室流式細胞分析儀設備規格參考如下表：
 
雷射光源
偵測器
(PMT)
Mirror(nm)
BP濾片(nm)
偵測範圍(nm)
(Emission)
可偵測螢光染料或螢光蛋白
488nm
藍光雷射
Blue A
735LP
780/60
750~810
 PE-Cy7
Blue B
655LP
670
>670
 PerCP, PerCP-Cy5.5
Blue D
556LP
585/42
564~606
 PE, PI, RFP
Blue E
502LP
530/30
515~545
 FITC, Alexa Fluor 488, GFP
640nm
紅光雷射
Red A
735LP
780/60
750~810
 APC-Cy7, APC-H7
690LP
730/45
708~753
 Alexa Fluor 700, iRFP713
Red B
—
670/30
655~685
 APC, Alexa Fluor 647, iRFP670
 
摘列市售符合上述條件之cell viability dye如下表，提供研究人員做為選購參考，相關問題可洽詢流式細胞儀室技術員 林倖如（分機：3949）。
 
☆ 適用於只做細胞表面染色不需內染的實驗之商品
 
廠牌
產品名稱
型號 / 規格
雷射
(nm)
Ex/Em
(nm)
相似螢光物質
檢測類型
Via-Probe
Red Nucleic Acid Stain		 565803 / 0.1 mL
565804 / 0.5 mL
633
631/651
 APC Nucleic acid binding
Via-Probe
Green Nucleic Acid Stain		 565799 / 0.1 mL
565802 / 0.5 mL
488
503/526
 FITC Nucleic acid binding
Pharmingen
Propidium Iodide Staining
Solution		 556463 / 2 mL
488
535/617
 PE Nucleic acid binding
Pharmingen
7-AAD		 559925 / 2 mL
488
546/647
 PerCP-Cy5.5 Nucleic acid binding
Propidium Iodide P1304MP / 100 mg
488
535/617
 PE Nucleic acid binding
TO-PRO-3 Iodide T3605 / 1 mL
633
642/661
 APC Nucleic acid binding
7-AAD A1310 / 1 mg
488
546/647
 PerCP-Cy5.5 Nucleic acid binding
SYTOX Green Nucleic Acid Stain S7020 / 250 uL
488
483/503
 FITC Nucleic acid binding
SYTOX Orange Nucleic Acid Stain S11368 / 250 uL
488
547/570
 PE Nucleic acid binding
SYTOX Deep Red Nucleic Acid
Stain		 S11380 / 50 uL
S11381 / 5 x50 uL
633
660/682
 APC Nucleic acid binding
SYTOX Green Dead Cell Stain S34860 / 1 mL
488
504/523
 FITC Nucleic acid binding
SYTOX Orange Dead Cell Stain S34861 / 1 mL
488
547/570
 PE Nucleic acid binding
SYTOX Red Dead Cell Stain S34859 / 1 mL
633
640/658
 APC Nucleic acid binding
SYTOX AADvanced Dead Cell Stain
Kit		 S10349 / 100 rxns
S10274 / 500 rxns
488
546/647
 PerCP-Cy5.5 Nucleic acid binding
CellTrace Calcein Green, AM C34852 / 20 x50 ug
488
495/515
 FITC Enzymatic
Propidium Iodide 75002 / 10mg
488
488/617
 PE Nucleic acid binding
7-AAD 75001.1 /200 tests
75001 / 500 tests
633
642/661
 APC Nucleic acid binding
     
☆ 對細胞固定液與打洞液具耐受性之商品
 
廠牌
產品名稱
型號 / 規格
雷射
(nm)
Ex/Em
(nm)
相似螢光物質
檢測類型
Horizon
Fixable Viability Stain 520		 564407 / 150 ug
488
498/521
 FITC Amine-reactive
Horizon
Fixable Viability Stain 570		 564995 / 150 ug
488
547/573
 PE Amine-reactive
Horizon
Fixable Viability Stain 620		 564996 / 100 ug
488
523/617
 PerCP-Cy5.5 Amine-reactive
Horizon
Fixable Viability Stain 660		 564405 / 100 ug
633
649/660
 APC Amine-reactive
Horizon
Fixable Viability Stain 700		 564997 / 100 ug
633
657/700
 Alexa Fluor 700 Amine-reactive
Horizon
Fixable Viability Stain 780		 565388 / 200 ug
633
759/780
 APC-Cy7 Amine-reactive
LIVE/DEAD Fixable Green stain L34969 / 80 tests
L23101 / 200 tests
L34970 / 400 tests
488
495/520
 FITC Amine-reactive
LIVE/DEAD Fixable Olive stain L34977 / 80 tests
L34978 / 200 tests
L34979 / 400 tests
488
480/557
 FITC Amine-reactive
LIVE/DEAD Fixable Far Red
stain		 L34973 / 80 tests
L10120 / 200 tests
L34974 / 400 tests
633
650/665
 APC Amine-reactive
LIVE/DEAD Fixable Scarlet stain L34986 / 80 tests
L34987 / 200 tests
L34988 / 400 tests
633
702/723
 Alexa Fluor 700 Amine-reactive
LIVE/DEAD Fixable Near-IR
(775) stain		 L34975 / 80 tests
L10119 / 200 tests
L34976 / 400 tests
633
750/775
 APC-Cy7 Amine-reactive
LIVE/DEAD Fixable Near-IR
(780) stain		 L34992 / 80 tests
L34993 / 200 tests
L34994 / 400 tests
633
633/785
 APC-Cy7 Amine-reactive
GloCell
Fixable Viability Dye Red 710		 75006.1 /100 tests
75006 / 500 tests
633
647/710
 Alexa Fluor 700 Amine-reactive
GloCell
Fixable Viability Dye Red 780		 75007.1 /100 tests
75007 / 500 tests
633
633/780
 APC-Cy7 Amine-reactive

嘉義長庚紀念醫院 第二共同實驗室 流式細胞儀室 2024年1月製表
 
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