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 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.
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 |
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 ML6640.
The ML6640 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.
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 ML6640 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 ML6640 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 is easily supported thanks to the Cloud connectivity of ML6600 platform.
- Predictive maintenance is supported both on-site and off-site.
Read more:
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.
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.