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流式細胞分選儀
細胞分選儀 FACSAria
分選儀 1 號機 BD FACSAria IIIu
4-laser, 13 colour cell sorting cytometer, Interchangeable emission filter set-up
分選儀 2 號機 BD FACSAria II
4-laser, 10 colour cell sorting cytometer, Interchangeable emission filter set-up
相關配備
(1) Nozzle : 70, 85 and 100 um
(2) Collection devices :
- for 2-way sorting (use 5 mL or 15 mL tubes for collecting cells), or
for 4-way sorting (using 5 mL tubes for collecting cells)
- ACDU (for 6-, 12-, 24-, 48- and 96-well plate)
上樣管 (自備)
可使用 PP 材質 (REF352053, 騰達行) 或 PS 材質 (REF352052 或 REF352235) 之 5mL Falcon 管上機,
不過 PP 材質較 PS 材質較不會造成細胞的吸附, 因此是較佳的選擇
光譜即時影像流式細胞分選儀 FACSDiscover™ S8 建置中
分選儀 3 號機 BD FACSDiscover S8 Cell Sorter with BD CellView™
5-laser, a spectral flow cytometer sorter with sort-capable image analysis
相關配備
(1) Nozzle : 85, 100, 130 um
(2) Collection devices :
- for 2~6-way sorting (use 1.5mL or 5mL tubes for collecting cells)
- ACDU (for 6-, 24-, 48- , 96- & 384-well plate sorting)
** 無法使用15mL離心管收集
** 85um nozzle可進行1~6向分選; 100um nozzle可進行1~4向分選; 130um nozzle可進行1~2向分選
上樣管 (自備)
可使用 PP 材質 (REF352053, 騰達行) 或 PS 材質 (REF352052 或 REF352235) 之 5mL Falcon 管上機,
不過 PP 材質較 PS 材質較不會造成細胞的吸附, 因此是較佳的選擇
流式細胞分析儀
細胞分析儀 FACSCalibur
分析儀 1~3 號機 BD FACSCalibur
2-laser, 4 colour analyzer, fixed emission filter set-up
上樣管 (自備)
僅可使用 PS 材質 (REF352052 或 REF352235, 騰達行) 之 5mL Falcon 管上機
清洗液
包含 Rinse, Clean 與 ddH2O 由本核心實驗室提供, 儀器使用完畢後, 請自行確實清洗乾淨
細胞分析儀 LSRFortessa
分析儀 4 與 5 號 機 BD LSRFortessa
5-laser, 18 colour analyzer, Interchangeable emission filter set-up
上樣管 (自備)
僅可使用 PS 材質 (REF352052 或 REF352235, 騰達行) 之 5mL Falcon 管上機
清洗液
包含 Rinse, Clean 與 ddH2O 由本核心實驗室提供, 儀器使用完畢後, 請自行確實清洗乾淨
雷射與集光濾鏡組態
分選儀1號機 FACSAriaIIIu
| Excitation Laser (nm) |
Dichroic mirror | Longpass Filter (nm) |
Bandpass Filter (nm) |
Fluorophores and other useful dyes |
|---|---|---|---|---|
| 488 | A | 655 LP | 695/40 | PerCP (87.4/63.3), PerCP-Cy5.5 (98.4/68.1), PE-Cy5.5 (76.9/38.5) |
| B | 502 LP | 530/30 | Alexa Fluor 488, FITC, GFP, Fluo-3 or Fluo-4 | |
| C | – | 488/10 | SSC | |
| 561 | A | 735 LP | 780/60 | PE-Cy7 (94.9/60.5) |
| B | 630 LP | 670/14 | PE-Cy5 (91.9/29.2), 7-AAD | |
| C | 600 LP | 610/20 | PE-Texas Red (94.6/30), PI DsRed (85.8/20.1) mCherry, |
|
| D | – | 582/15 | dTomato, PE(94.3/~30), RFP, Cy3 DsRed (85.8/22.3), |
|
| 633 | A | 735 LP | 780/60 | APC-Cy7, APC-H7, Alexa Fluor® 750 |
| B | – | 660/20 | APC, Alexa Fluor® 647, Cy5, MitoProbe™ DiIC1(5), CellTrace™ Far Red DDAO-SE, SYTOX® Red |
|
| 405 | A | 630 LP | 660/20 | Brilliant Violet650, Qdot 655 |
| B | 600 LP | 610/20 | Brilliant Violet605, Qdot 605 | |
| C | 556 LP | 585/42 | Brilliant Violet570, Qdot565 | |
| D | 502 LP | 510/50 | Brilliant Violet510, AmCyan, Horizon V500, | |
| E | – | 450/40 | Pacific Blue, Brilliant Violet421, Alexa Fluor® 405, Horizon V450, |
** 加註刪除線者表示儀器組態中有2個或2個以上的位置可以收集到該螢光訊號，加註刪除線 的位置非該螢光試劑之最佳收集位置，建議以未加註刪除線之位置收集其螢光訊號。
分選儀2號機FACSAria II
| Excitation Laser (nm) |
Dichroic mirror | Longpass Filter (nm) |
Bandpass Filter (nm) |
Fluorophores |
|---|---|---|---|---|
| 488 | A | 735 LP | 780/60 | PE-Cy7 (61.8/60.5) |
| B | 655 LP | 695/40 | PerCP (87.4/63.3), PerCP-Cy5.5 (98.4/68.1), PE-Cy5.5 (76.9/38.5), |
|
| C | 610 LP | 616/23 | PE-Texas Red (94.6/35), PE-Alexa Fluor® 610 |
|
| D | 556 LP | 585/42 | DsRed (40/59.7), PE (61.6/70.4), Cy3 |
|
| E | 502 LP | 530/30 | Alexa Fluor 488, FITC, GFP, Fluo-3 or Fluo-4, Calcein, TO-PRO®-1, YO-PRO®-1, CFSE, Rhodamine123… | |
| F | – | 488/10 | SSC | |
| 561 | A | 735LP | 780/60 | PE-Cy7 |
| B | 630LP | 670/14 | PE-Cy5.5 | |
| C | 600LP | 610/20 |
PE-Texas Red, mCherry DsRed (85.8/20.1), |
|
| D | - | 582/15 |
dTomato, RFP, Cy3, PE DsRed (85.8/22.3) |
|
| 633 | A | 735 LP | 780/60 | APC-Cy7, APC-H7, Alexa Fluor® 750 |
| B | - | 660/20 | APC, Alexa Fluor® 647, Cy5 | |
| 375 (NUV) | A | 610LP | 670LP | Hoechst Red |
| B | – | 450/40 | Hoechst Blue33258 (57.9/38.3), DAPI (79.8/36.9) |
|
| – | 502LP | 530/30 | 未使用 | |
| – | - | 450/20 | 未使用 |
分選儀3號機 FACSJazz
| Excitation Laser (nm) | Fluorescence Channel |
Bandpass Filter (nm) |
Color | Fluorophores and other useful dyes |
|---|---|---|---|---|
| 488 | A | 710/50 | Red | PerCP-Cy5.5, PE-Cy5.5 |
| B | 530/40 | Green | Alexa Fluor 488, FITC, GFP, Fluo-3 or Fluo-4, Calcein, TO-PRO®-1, YO-PRO®-1, CFSE, Rhodamine123… | |
| 561 | C | 750LP | Dark Red | PE-Cy7 |
| D | 610/20 | Orange | mCherry, PE-Texas Red, PI | |
| E | 585/29 | Yellow | dTomato, DsRed, PE, Cy3 | |
| 640 | F | 660/20 | Red | APC, Alexa Fluor 647 |
請注意:
- 以本儀器進行分析時，強烈建議勿使用PerCP螢光染劑。
如有疑問，請洽本核心人員。
分析儀1-3號機 FACSCalibur
| Excitation Laser (nm) |
Fluorescence Channel |
Bandpass Filter (nm) |
Color | Fluorophores and other useful dyes | |
|---|---|---|---|---|---|
| 488 | FL1 | 530/30 | Green | Alexa Fluor 488, FITC, GFP, |
Fluo-3 or Fluo-4, Calcein, CFSE, GFP, Rhodamine123, TO-PRO®-1, YO-PRO®-1, Click-i EdU Alexa Fluor 488, JC-1 or DiOC2(3), SYTOX® Green, DyeCycle™ Green LIVE/DEAD® Fixable Green Dead Cell Stain |
| FL2 | 585/42 | Yellow/ Orange |
PE, DsRed, |
PI, Fura Red, DyeCycle™ Orange, JC-1 or DiOC2(3), SNARF® (low pH), pHrodo™ dye |
|
| FL3 | 670LP | Dark Red | PE-Cy5.5, PE-Cy7 PerCP, PerCP-Cy5.5, PE-Alexa Fluor 700, |
PI, 7-AAD, SNARF® (high pH), DiOC2(3), LIVE/DEAD® Fixable Red Dead Cell Stain |
|
| 635 | FL4 | 661/16 | Red | APC, Alexa Fluor 647, |
Click-i EdU Alexa Fluor 647, LIVE/DEAD® Fixable Far Red Dead Cell Stain, TO-PRO®-3, SYTOX® Red, MitoProbe™ DiIC1(5), CellTrace Far Red DDAO-SE |
** 若需要，本表所列出的螢光試劑當然亦可適用於其他具有類似雷射/集光濾 鏡組態的流式細胞儀。
分析儀4與5號機 LSRFortessa
| Excitation Laser (nm) |
Dichroic mirror | Longpass Filter (nm) |
Bandpass Filter (nm) |
Fluorophores & other useful dyes |
|---|---|---|---|---|
| 355 (UV) |
A | 690LP(暫) | 740/35(暫) | BUV737, (或換 BUV803) |
| B | 505LP(暫) | 530/30(暫) | DAPI-B, (或換 Hoechst Red, BUV661, BUV496) | |
| C | 450/50(暫) | DAPI-C, Hoechst Blue, (或換 BUV395) | ||
| 替 換 用 濾 鏡 |
770LP | 820/60 | BUV803 | |
| 690LP | 740/35 | BUV737 | ||
| 505LP | 670LP | Hoechst Red | ||
| 630LP | 670/25 | BUV661 | ||
| 450LP | 515/30 | BUV496 | ||
| 450/50 | Hoechst Blue | |||
| 379/28 | BUV395 | |||
| 405 (Violet) |
A | 685LP | 710/50 | BV711 (* 若需要, 可更換濾鏡為 780/60, 使用染劑 BV786) |
| B | 630LP | 655/8 | BV650, Qdot 655 | |
| C | 595LP | 605/12 | BV605, Qdot 605 | |
| D | 545LP | 560/20 | BV570, Qdot565 | |
| E | 475LP | 525/50 | BV510, AmCyan, Qdot525, Horizon V500, SYTOX Blue, Indo-1 (low Ca2+), CFP |
|
| F | - | 450/50 | BV421, Pacific Blue, eFluor 450, Horizon V450, Alexa Fluor 405, DyeCycle Violet, TaqBFP/BFP |
|
| 488 (Blue) |
A | 685LP | 695/40 | BB700, PerCP , PerCP-Cy5.5, PI |
| B | 505LP | 530/30 | Alexa Fluor 488, BB515, FITC, GFP, YFP, YO-PRO®-1, Fluo-3或Fluo-4, Calcein, TO-PRO®-1, JC-1, CFSE… |
|
| C | - | 488/10 | SSC | |
| 561 (yellow green) |
A | 750LP | 780/60 | PE-Cy7(YG) |
| B | 635LP | 670/30 | PE-Cy5 (YG) 7-AAD-B(YG), PI-B(YG) |
|
| C | 600LP | 610/20 | PE-Texas Red(YG), PE-CF594(YG), mCherry(YG), Alexa Fluor594(YG), PI-C(YG), |
|
| D | 582/15 | PE(YG), dTomato(YG), DsRed(YG), RFP(YG), Cy3(YG) | ||
| 640 (Red) |
A | 750LP | 780/60 | APC-Cy7, APC-H7, Alexa Fluor® 750 |
| B | 710LP | 730/45 | Alexa Fluor 700, Cy5.5 | |
| C | - | 670/14 | APC, Aexa Fluor® 647, Cy5, MitoProbe™ DiIC1(5), SYTOX Red |
** 欲使用355nmUV雷射的研究人員請注意: 有三組濾鏡位置 (A、B與C) 可供分析使用，
濾鏡為抽換式的，需視實際需求抽換合適之濾鏡組，預約與使用前，請務必先與管理人員確認。
