Flow Cytometry

Flow Cytometry

Flow cytometry is a widely used method in biomedical research and increasingly also in clinical diagnostics. It is a powerful and rapid technique to analyze physical and chemical properties of single cells or particles as they are suspended in liquid and pass in a narrow line across laser beams.

Introduction

Flow cytometry is a widely used method in biomedical research and increasingly also in clinical diagnostics. It is a powerful and rapid technique to analyze physical and chemical properties of single cells or particles as they are suspended in liquid and pass in a narrow line across laser beams. Fluorescence together with scattered laser light is then filtered, detected, and analyzed on the other side of the sample. In addition to analysis, many flow cytometers can also sort and purify cell populations of interest for downstream analysis based on the identified properties of cells or particles.

Modulight's solution for Flow Cytometry

Modulight's wavelength range

Modulight offers multiple laser lines for exciting different fluorescent labels, each with its own absorption and emission characteristics, increasing the versatility and flexibility of the system. The most common wavelengths based on the antibody labels used in the biosciences field include 405 nm, 445 nm, 488 nm, 532 nm, 561 nm, 633 nm, 640 nm, 660 nm, and 808 nm. Modulight covers the wavelength range of UV to 2000+ nm, with many options to choose from for the new upcoming dyes in the biomedical field.

Some examples of the most common supported fluorophores

Thanks to Modulight’s inhouse capabilities to manufacture lasers between UV and 2000+ nm, we can match these and many more fluorophore types, even uncommon and new ones.

Fluorophore Absorption (nm) Emission (nm)
Alexa Fluor 405 401 442
Pacific Blue 404 455
AmCyan 457 489
PerCP 477 678
EGFP 489 510
FITC 495 519
Alexa Fluor 488 499 520
7-AAD 543 647
Cy3 548 561
PE-Cy5 565 665
PE 566 576
PE-Cy7 566 778
Alexa Fluor 568 577 603
Alexa Fluor 594 590 618
APC 650 660
Alexa Fluor 647 653 668
Alexa Fluor 700 696 719
APC-Cy7 756 779
Alexa Fluor 790 782 805
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We support laser wavelengths from UV to 2000+ nm

Fluorophore Absorption (nm) Emission (nm)
Indo-1 hi 223 403
Riboflavin 266 531
Cascade Yellow 277 548
Emerald 300 289 530
QDot 525 300 525
QDot 545 300 543
QDot 565 300 564
QDot 585 300 588
QDot 605 300 603
QDot 625 300 621
QDot 655 300 654
QDot 705 300 702
QDot 800 300 792
Fura-2 hi 336 505
FVS440UV 336 435
Alexa Fluor 350 342 441
Indo-1 lo 345–346 479
AMCA 346 442
BUV395 347 393
BUV496 350 491
BUV563 350 560
BUV615 350 661
BUV661 350 653
BUV737 350 732
PI 351 617
BUV805 352 803
Hoechst33258 352 455
DAPI 357–359 461
Methyl Coumarin 360 448
Marina Blue 364 460
Fura-2 lo 367 515
DyeCycle Violet 369 437
FVS575V 396 572
Cascade Blue 398 420
Calcein Violet AM 400 452
Pacific Orange 400 551
Alexa Fluor 405 401 442
BV510 402–407 510
FVS450 404 448
Pacific Blue 404 455
V450 405 450
BV711 405–408 710
BV570 407 573
BV605 407 601
BV650 407 646
BV786 407 786
BV750 407–408 745
BV421 408 421
V500 415 499
FVS510 418 509
Lucifer Yellow 428 544
Alexa Fluor 430 431 540
Fura Red Hi 434–435 666
ECFP 435 477
BV480 436 476
SYTOX Blue 445 470
AmCyan 457 489
DiA 457 586
Fura Red Lo 471 672
BB630P2 476 613
BB660P2 476 658
BB700 476 695
BB755P 476 758
PerCP 477 678
PerCP-Cy5.5 480–482 676
BB790P 485 784
DiO 489 506
EGFP 489 510
CF488A 489–490 516
BB515 490 513
FVS520 491 517
YO-PRO-1 491 506
YOYO-1 491 508
Calcein pH 9 493 514
Fluo-4 494 516
ZsGreen 494 516
FITC 495 519
Fluoro-Emerald 495 523
RB545 496 544
Rho110 497 520
Oregon Green 498 526
RB780 498 781
SYBR Green 498 522
Alexa Fluor 488 499 520
BODIPY 502 511
CyQUANT DNA 502 523
PicoGreen 502 522
Alexa Fluor 500 503 525
SYTOX Green 503 524
DyeCycle Green 506 534
Fluo-3 506 527
Na-Green 506 531
Magnesium Green 507 531
Rho123 507 529
Via-Probe Green 511 534
TOTO-1 514 531
EYFP 515 528
TO-PRO-1 515 531
DyeCycle Orange 519 563
6-JOE 520 548
FVS620 522 617
EB 523 604
Eosin 525 546
Alexa Fluor 532 528 552
BODIPY R6G 528 547
mBanana 540 553
7-AAD 543 647
SYTOX Orange 547 569
Cy3 548 561
mOrange 548 562
SNARF-1 (pH6) 549 586
DiI 551 569
CoralHue mk02 552 565
Kusabira-Orange 552 565
DsRED 553 585
FVS570 553 578
TMRho 553 581
CF555 554 568
dTomato 554 581
Cy3B 559 571
Nile Red 559 637
RY586 564 588
BYG584P 563–564 582
PE-Cy5 565 665
PE-Cy5.5 565 693
PE 566 576
PE-CF594 566 612
PE-Cy7 566 778
PE-Texas Red 566 616
mTangerine 568 585
Rhodamine Red 573 591
mStrawberry 574 596
SNARF-1 (pH9) 576 640
Cy3.5 576–579 589
Alexa Fluor 568 577 603
MitoTracker Red 578 598
mCherry 587 610
mPlum 587 650
Alexa Fluor 594 590 618
JC-1 593 595
Texas Red 595 613
mRaspberry 597 624
YOYO-3 612 631
YO-PRO-3 613 629
Phycocyanin 617 646
Alexa Fluor 633 631 647
Via-Probe Red 631 651
SYTOX Red 640 658
TO-PRO-3 642 657
TOTO-3 642 661
Cy5 647 662
DRAQ5 647 683
DiD 648 670
APC 650 660
CF647 652 668
Alexa Fluor 647 653 668
APC-R700 653 706
FVS660 656 670
FVS700 656 697
Alexa Fluor 660 663 691
CF660 666 685
Alexa Fluor 680 679 702
CF680 681 689
R718 694–697 715
Alexa Fluor 700 696 719
DiR 750 782
Alexa Fluor 750 752 779
Cy7 753 775
CF750 755 779
APC-Cy7 756 779
FVS780 757 778
APC-H7 759 782
CF770 770 797
Alexa Fluor 790 782 805

Modulight solution for Flow Cytometry

Modulight is specialized to provide whole fluorescence sensing solution for flow cytometry, with all the needed wavelengths in a compact, easy-to-integrate single package, the ML6600.

ML6600 for flow cytometry
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The ML6600 for flow cytometry provides focused, noise-free, and monochromatic illumination for rapid detection and analysis as the samples are passing through the flow cell.

The system delivers a focused, noise-free, and monochromatic output, offering up to four selectable wavelengths: 349 nm (UV), 405 nm (violet), 488 nm (blue), 561 nm (yellow), and 638 nm (red). Output beams have flat-top profiles and are pre-configured into a beam ladder.

The ML6600 for Flow Cytometry features a built-in CCD camera for beam monitoring and the beams can be individually steered either remotely via the Modulight Cloud or automatically with the AI-based beam alignment system. The ML6600 platform supports preventive maintenance with predictive analytics and machine learning algorithms, powered by Modulight Cloud.

Having multiple laser wavelengths in flow cytometry increases the versatility and flexibility of the system: multiple fluorescent targets can be detected within the same experiment, with more freedom for fluorophore selection. Modulight provides complete optical subsystems for flow cytometry, including detection optics for all of the most important fluorescent dyes in a compact and cost-effective package, both off-the-shelf and tailored, based on customers’ requirements

The multi-wavelength ML6600 integrates all the basic optical elements needed in flow cytometers into one single box, providing the customer a single optical arrangement solution with all the most important wavelengths in a compact package. These elements include the illumination subsystem, fine-tuned optics, and a detection system (optional).

Benefits

  • The ML6600 for flow cytometry introduces a high level of integration and miniaturization of the most important elements in flow cytometers.
  • Wide range of wavelengths and laser technologies supported, from UV to 2000+ nm, for existing and new fluorophore excitation.
  • From mW to W level of output power.
  • AI-based automated beam alignment of each individual laser beam.
  • Beam shaping optics allow for differently shaped output beams to illuminate the flow cell according to the customer’s specifications.
  • Maintenance service (i.e., annual calibrations) and predictive maintenance of the ML6600 is easily supported thanks to the Cloud connectivity of ML6600.
  • Predictive maintenance is supported both on-site and off-site.

ML6600 is a completely customizable and easy-to-integrate laser platform.

Let's schedule a call and find the perfect integrated solution for you!

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Application note: Flow Cytometry

Flow cytometers can have from one to multiple laser lines for exciting different fluorescent labels each with own absorption and emission characteristics. The laser wavelengths are chosen based on the most commonly used antibody labels in biosciences: 405 nm, 445 nm, 488 nm, 532 nm, 561 nm, 633 nm, 640 nm, 660 nm, and 810 nm. Even with one laser, several different fluorescent labels with overlapping absorption spectra can be excited; however, having multiple laser wavelengths increases the versatility and flexibility of the system: multiple fluorescent targets can be detected within the same experiment, with more freedom for fluorophore selection.

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ML6600: Cloud-powered laser solution platform 

ML6600 is a compact, easy-to-integrate laser solution that can be flexibly tailored to various applications, such as Flow Cytometry, Semiconductor Metrology, Quantum Computing, Diagnostics, Genetics, and various medical applications.

It’s well suited for medical system integrators, as it’s designed in accordance with IEC 60601-1 norm. This will speed up the certification process. The lasers and the system are manufactured in ISO13485 certified fab.

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