Flow Cytometry Resources

1. ZE5 cell analyzer with five lasers:

1. Coherent OBIS BSO 355nm 50mW laser with:

        A. 690DLP, 700LP (BUV737)

B. 560DLP, 670/30BP (BUV661, Hoechst 33342-Red, PI)

C. 493DLP, 525/50BP (BUV496, BUV 563)

D. 405DLP, 447/60BP (Alexa350, DAPI, Hoechst 33342-Blue, Indo-1 low, -Ca++)

E. None, 387/11BP (BUV395, Indo-1 high, +Ca++)

2. Coherent OBIS BSO 405nm 100mW laser with:

        A. 752DLP, 750LP         (BV786)

B. 690DLP, 720/60BP (BV711)

C. 640DLP, 670/30BP (BV650)

D. 581DLP, 615/24BP (BV605)

E. 493DLP, 525/50BP (AmCyan, BV510, Cascade Yellow, Pacific Orange)

F. 439DLP, 460/22BP (Pacific Blue, Horizon BV450, DAPI)

G. None, 420/10BP (Cascade Blue, BV421, Alexa405)

3. Coherent OBIS BSO, 488nm 100mW laser with:

        A. 752DLP, 750LP (PE-Cy7, PE-Alexa750)

        B. 629DLP, 692/80BP (PE-Cy5, PerCP-Cy5.5, PerCP, PE-Cy5.5,  PE-Alexa647)

C. 558DSP, 593/52BP (PE, PE-Texas Red, PI)

D. 493DLP, 525/35BP (eGFP, FITC, Alexa488)

E. None, 488/10BP (SSC)

4. Coherent OBIS BSO 561nm 50mW laser with:

        A. 750DSP, 750DLP (PE-Cy7, PE-Alexa750)

B. 690DLP, 720/60BP (PE-Cy-5.5)

C. 652DLP, 670/30BP (PE-Alexa647, PE-Cy5)

D. 629DLP, 640/20BP (mPlum)

E. 600DLP, 615/24BP (mCherry, PE-Texas Red, PI, Texas Red)

F. 581DLP, 589/15BP ()

G. None, 577/15BP (dTomato, dsRed, PE)

5. Coherent OBIS BSO 640nm 100mW laser with:

        A. 800DLP, 800LP ()

B. 752DLP, 775/50BP (APC-Cy7, APC-H7, APC-Alexa750)      

C. 690DLP, 720/60BP (Cy5.5, Alexa680, Alexa700)

D.  None, 670/30BP (APC, Cy5, Alexa647)

 

2. LSRII-new cell analyzer with five lasers:

1. Coherent Sapphire Solid state 488nm, 20mW laser with:

        A. 685LP, 710/50BP (PerCP-Cy5.5)

        B. 505LP, 525/50BP (Alexa488, FITC, GFP, CFSE, YFP)

        C. None, 488/10BP (SSC)

2. Coherent COMPass YellowGreen 561nm, 25mW laser with:

       A. 740LP, 780/60BP (PE-Cy7, PE-Alexa750)

        B. 685LP, 710/50BP (PE-Alexa680, PE-Cy5.5)

        C. 635LP, 660/20BP (PE-Cy5, PE-Alexa647, 7AAD)

        D. 600LP, 610/20BP (PI, Alexa594, HCred, mCherry, mRFP1, PE-Texas Red, Alexa568, PE-CF594)

        E. 570LP, 582/15BP (DsRed, PE, DsRed2, dTomato,Cy3)

3. JOS Uniphase 1344p HeNe Red 633nm, 40mW laser with :

        A. 740LP, 780/60BP (APC-Cy7, APC-Alexa750, APC-H7)

        B. 685LP, 710/50BP (Alexa680, APC-Cy5.5, Alexa700)

        C. None, 670/30BP (APC, Alexa647, Cy5)

4. Coherent Vioflame solid state Violet 407nm, 50mW laser with:

        A. 740LP, 780/60BP (Qdot800)

        B. 685LP, 705/70BP (Qdot705)

        C. 565LP, 610/20BP (Qdot605, BV605)

        D. 505LP, 525/50BP (Alexa430, CFP, Qdot565,AmCyan, HorizenV500)

        E. None, 450/50BP (Alexa405, Cascade-Blue, PacificBlue, eFluor450, HorizenV450, BV421, CFP)

5. LightwaveXcyte solid state UV 355nm, 60mW laser with:

       A. 505LP, 530/30BP (Hoechst Red, Indo1 blue)

       B. none, 450/50BP (Alexa350, Hoechst, Indo1 violet, DAPI)

3. LSRII cell analyzer with three lasers:

1. Coherent Sapphire Solid state 488nm, 20mW laser with:

        A. 755LP, 780/60BP (PE-Cy7)

        B. 685LP, 695/40BP (PerCP-Cy5.5, PerCP)

        C. 655LP, 670/14BP (PE-Cy5, Nile red, 7AAD)

        D. 600LP, 610/20BP (PE-Texas Red, Propidiume Iodide, Diyhdroethidium, PE- CF594)

        E. 550LP, 575/26BP (PE, PI, DsRed, Dil)

        F. 505LP, 530/30BP (FITC, Alexa488, GFP, SYBR green, YFP, CFSE)

        G. None 488/10BP (SSC)

 2. JDS Uniphase HeNe 633nm, 17mW laser with:

        A. 755LP, 780/60BP (APC-Cy7, APC-H7, Alexa750)

        B. 710LP, 730/45BP (Alexa700)

        C. None, 660/20BP (APC, Cy5, DiD)

  3. Point Source Violet Solid State 407nm, 25mW laser with:

        A. 505LP, 525/50BP (Alexa430, AmCyan, HorizenV500, CFP)

        B. None, 450/50BP (Pacific Blue, Alexa405, Marine Blue, CFP, coumarine Hcc2, eFluor450, BV421)

4. FACSAria-FUSION with five lasers.

The blue, yellow-green and red can be used either with the violet or the nUV

1. 488nm – Blue – 50mW (Coherent Sapphier):

        A. 655LP, 695/40BP (695/40-B, PerCp-cy5.5 ,PerCP)

        B. 502LP, 530/30BP (530/30-B, FITC, Alexa 488, Brilliant Blue515 , GFP, CFSE, YFP, SYBR Green)

2. 561nm - Yellow Green – 50 – mW (Coherent Sappier):

        A. 735LP, 780/60BP (780/60-YG, PE-Cy7, PE-Alexa750)

        B. 685LP, 710/50BP (710/50-YG, PE-Cy5.5, PE-Alexa680)

        C. 630LP, 670/14BP (670/14-YG, PE-Cy5, 7AAD, PE-Alexa647)

        D. 600LP, 610/20BP (610/20-YG, PI, PE-CF594, PE-TxRed, Alexa594, mCherry, mRFP1,Alexa568)

        E. None, 582/15BP (582/15-YG, PE, DsRed, dTomato, Cy3)

3. 640 nm – Red – 100mW (Coherent Obis):

        A. 755LP, 780/60BP (780/60-R, APC-Cy7, APC-H7, APC-Alexa750)

        B. 690LP, 730/45BP (730/45-R, Alexa700)

        C. None, 670/30BP (670/30-R, APC, Alexa647, Cy5)

4. 405nm –Violet – 85mW (Point Source Iflex 2000):

        A. 750LP, 780/60BP (780/60-V, BV786, Qdot800)

        B. 690LP, 710/50BP (710/50-V, BV711, Qdot705)

        C. 630LP, 660/20BP (660/20-V, BV660)

        D. 595LP, 610/20BP (610/20-V, BV605, Qdot605)

        E. 505LP, 525/50BP (525/50-V, AmCyan, Horizon V500, BV510, Alexa430)

        F. None, 450/50BP (450/50-V, Pacific Blue, BV421, Alexa405, Cascade-Blue, eFluor450, CFP, DAPI-Violet)

5. 375nm - nUV - 7mW (Fabrinet):

        A. 610LP, 670LP (670LP-nUV, Hoechst Red)

        B. None, 450/20BP (450/20-nUV, DAPI-nUV, Hoechst Blue)

5. FACSAriaIII cell sorter with four lasers:

1. Coherent Blue 488nm 20mW laser with:

        A. 655LP, 695/40BP (PerCp-Cy5.5, PerCP)

        B. 502LP, 530/30BP (FITC, CFSE, GFP, YFP, Alexa488)

        C. None, 488/10BP (SSC)

2. Coherent Yellow Green 561nm 40mW laser with:

        A. 735LP, 780/60BP (PE-Cy7, PE-Alexa750)

        B. 685LP, 710/50BP (PE-Cy5.5, PE-Alexa680)

        C. 630LP, 670/14BP (PE-Cy5, PE-Alexa647, 7AAD)

        D. 600LP, 610/20BP (PI, PE-TxRed, Alexa594, mCherry, HCred, mRFP1, PE-CF594)

        E. None, 582/15BP (PE, DsRed, Alexa568, dTomato, DsRed2)

3. JDSU HeNeRed 633 nm 20mW laser with:

        A. 755LP, 780/60BP (APC-Cy7, APC-Alexa750, APC-H7)

        B. 690LP, 730/45BP (APC-Cy5.5, Alexa700)

        C. None, 660/20BP (APC, Cy5, Alexa647)

4. Coherent Violet 407nm 50mW with:

        A. 735LP, 780/60BP (QDOT800)

        B. 690LP,  710/50BP (BV711, Qdot705, eFluor 700NC)

        C. 630LP,  660/20BP (BV650, Qdot655, eFluor 650NC, eVolve650)

        D. 595LP, 610/20BP  (BV605, Qdot605, eFluor 605NC, eVolve605)

        E. 502LP, 510/50BP (Alexa430, CFP, QD 525, AmCyan, Horizen V500, CFP)

        F. None, 450/40BP (Alexa405, Pacific Blue, Cascade Blue, eFluor 450, Horizen V450, BV421, CFP)

6. SORP FACSAriaII cell sorter with five lasers:

1. UV Genesis 355nm 65mW laser with:

        A. 505LP, 530/30BP (Hoechst red, Indo-1 blue)

        B. None, 450/50BP (Alexa350, Hoechst, Indo-1 Violet, DAPI-UV)

2. Violet CUBE 405nm 100mW with:

        A. 750LP, 780/60BP (Qdot800)

        B. 670LP, 705/70BP (Qdot705)

        C. 595LP, 605/40BP (Qdot605, Brilliant Violet605)

        D. 505LP, 525/50BP (Alexa430, AmCyan, CFP, Qdot525, Horizen V500, CFP)

        E. None, 450/50 (Alexa405, Cascade Blue, Pacific Blue, DAPI- Violet, CFP,Horizen V450, BV421)

3. Blue Sapphire 488nm 100mW with:

        A. 685LP, 710/50BP (PerCp-Cy5.5)

        B. 505LP, 525/50BP (Alexa488, FITC, GFP, CFSE, YFP)

        C. None, 488/10BP (SSC)

4. YellowGreen Compass 561nm 110mW with:

        A. 750LP, 780/60BP (Pe-Cy7, PE-Alexa750)

        B. 685LP, 710/50BP (PE-Alexa680, PE-Cy5.5)

        C. 635LP, 660/20BP (PE-Cy5, PE-Alexa647)

        D. 600LP, 610/20BP (PE-Texas Red, PI, Alex 594, HCred,mCherry, mRFP1, PE- CF594)

        E. None, 582/15BP (DsRed, PE, DsRed2-Alexa568, dTomato)

5. Red CUBE 640nm 40mW with:

        A. 750LP, 780/60BP (APC-Cy7, APC-Alexa750, APC-H7)

        B. 685LP, 710/50BP (Alexa680, Alexa700, APC-Cy5.5)

        C. None, 670/14BP (APC, Alexa647, Cy5)

7. FACSAriaII cell sorter with three lasers:

1. Coherent Sapphire Solid state 488nm, 20mW laser with:

        A. 735LP, 780/60BP (PE-Cy7, aposense)

        B. 655LP, 695/40BP (PE-Cy5, PerCp, PerCp-Cy5.5, PI, 7AAD, Cy-Chrome)

        C.  595LP, 610/20BP (PE-Texas Red, HcRed1, Propidium Iodide, PE-CF594)

        D. 550LP, 575BP (DsRed, PE, Cy3, DsRed2, RFP)

        E. 502LP, 530/30BP (Alexa488, FITC, GFP, CFSE, SYBR Green, YFP)

        F. None, 488/10BP (SSC)

2. JDS Uniphase HeNe 633nm, 17mW with:

        A. 735LP, 780/60BP (APC-Cy7, APC-H7, APC-Alexa750)

        B. None, 660/20BP (APC, Alexa647, Cy5, Did)

3. Point Source Violet Solid State 407nm, 25mW with:

        A. 502LP, 530/30BP (Alexa430, AmCyan.,CFP,Horizen V500)

        B. None, 450/40BP (CFP, DAPI, Hoechst Blue, Marina Blue, Pacific Blue, Horizen V450, BV421)

8. FACSMelody cell sorter with three lasers:

1. Violet 405nm diode laser, 40mW with:

        A. 755LP, None (BV786)

        B. 500LP, 528/45BP (V500, BV510, Pacific Orange, AmCyan)

        C. None, 448/45BP (V450, Pacific Blue, DAPI, BV421)

2. Blue 488nm diode laser, 20mW with:

        A. 752LP, 783/56BP (PE-Cy7, PE-Alexa750)

        B. 665LP, 700/54BP (PE-Cy5.5, PerCP, PerCP-Cy5.5, 7-AAD, BB700)

        C. 560LP, 586/42BP (PE, PI)

        D. 507LP, 527/32BP (FITC, GFP, BB515, Alexa488, CFSE)

        E. None, 488/15BP (SSC)

3. Red 640nm diode laser, 40mW with:

        A. 752LP, 783/56BP (APC-Cy7, APC-Alexa750, APC-H7)

        B. None, 660/10BP (APC, Alexa647, Cy5)

9. ImageStreamX Flow Imager system with 5 lasers:

1. Excitation lasers:

        A. Violet 405 nm, 120mW

        B. Blue 488 nm 100 mW

        C. Yellow-Green 561 nm, 200 mW

        D. Red 642 nm 150 mW

        E. Darkfield laser 785 nm (SSC).

2. Emission channels (nm):

        A. 430-480: Brightfield

        B. 480-560: FITC, GFP, AF488, YFP

        C. 560-595: PE, Cy3, AF546, AF555

        D. 595-660: RFP, PI, mCherry, AF568

        E. 660-745: PerCP, PE-Cy5, PE-AF647

        F. 745-800: PE-Cy7, PE-AF750, SSC

        G. 430-505: DAPI, Hoechst, Pacific blue, CFP, A405

        H. 505-570: PacOrange, AF430, QD525

        I. 570-595: Brightfield

        J. 595-640: QD625, eFluor625

        K. 640-745: APC, Cy5, AF647, Draq5, PerCP

        L. 745-800: QD800, APC-Cy7,APC -AF750,Cy7, AF750

3. The ImageSreamX contains:

        A. Two cameras, 12 Imaging Channels

        B. 20X,40X and 60X magnification

        C. Extended Depth of Field optics

        D. Single color brightfield

        E. Autosampler for 96 well plates

        F. Data Analysis Workstation with IDEAS software.

10. ImageStream-NEW Flow Imager system with 5 lasers

1. Excitation lasers:

        A. Violet 405 nm 120mW

        B. Blue 488nm 400 mW

        C. Yelow-Green 561nm 200 mW

        D. Red 642 nm 150 mW

        E. Darkfield laser 785 nm (SSC)

2. Emission channels (nm):

        A. 430-480: Brightfield

        B. 480-560: FITC, GFPm AF488, YFP

        C. 595-660: RFP, PI, mCherry, AF568

        D. 660-745: PerCP, PE-Cy5, PE-AF647

        E. 745-800: PE-Cy7, PE-AF750, SSC

        F. 430-505: DAPI, Hoechst, Pacific blue, CFP, A405

        G. 505-570: PacOrange, AF430, QD525

        H. 570-595: Brightfield

        I. 595-640: QD625, eFluor625

        J. 640-745: APC, Cy5, AF647, Draq5, PerCP

        K. 745-800: QD800, APC-Cy7, APC-AF750, Cy7,AF750

3. The ImageStreamX contains:

        A. Two cameras, 12 Imaging Channels

        B. 20X, 40X and 60X magnification

        C. Single color brightfield

        D. Data Analysis Workstation with IDEAS software

11. CyTOF2 Mass Cytometer upgraded to Helios
12. EVOS FL Auto 2 Imaging System

        A. Fully-automated inverted multi-channel fluorescence and transmitted light imaging system.

        B. Equipped with on stage incubator with CO2 and temperature control.

        C. Available lenses: 4X, 20X.

        D. Available channels - brightfield and green (GFP, FITC, AF488)

13. Additional Equipment
  • A PC with FlowJo, FCS-express and Modfit for off line analysis.
  • An_Heraeus_Labofuge 400 Centrifuge.
  • An HERA cell Heraeus_Incubator.
  •  A NUAIR Biological Safety Cabinet.
  •  Floor grounding to avoid static electricity.
  • Individual air conditioning with positive pressure. Temperature maintained between 22-24C.
  •  Central air pressure outlet allowing up to 80 PSI.
  • Vacuum pump.
  • Fluorescent room lighting and additional rheostat control.
  • Walls and sinks with non-reflective surfaces.

   

Safety:
    A danger sign at the entry to the laboratory.
    A red light to be turned on when lasers are aligned.
    A main disconnect for the lasers located near the operator.
    Electric locking of the room with magnetic cards to enter