Danube 

Fluorescence Lifetime Flow Cytometer


In cell biology and cancer research, there is often the need to measure cellular processes, protein function, protein-protein interactions, or molecular transport with subcellular resolution. Fluorescence lifetime is a powerful tool that can provide this information. Traditionally, fluorescence lifetime techniques (such as FRET-FLIM) have been carried out on imaging platforms; however, the low throughput of microscopy severely limits the resulting efficiency of analysis.

The Danube combines many of the benefits of FLIM analysis with the inherent high throughput of flow cytometry. The most advanced fluorescence lifetime flow cytometer on the market, it provides direct, time-domain analysis of fluorescence lifetime, with the ability to measure multi-exponential decay on a cell-by-cell basis at a throughput of up to 5,000 cells/second

Danube measurement of multiexponential fluorescence lifetime decay
of a mixture of PyBlue (
t1) and a blue-emitting quantum dot (t2).


The Danube works by generating extremely short interactions between the interrogating laser light and the cells in the sample. Each cell is probed dozens of times, with each excitation event lasting 4 ns or less. This capability, unique in flow cytometry, results in subnanosecond time resolution of fluorescence lifetime decay values, and the ability to measure lifetime changes in most of the fluorophores and fluorescent proteins in common use.

Working directly in the time domain, the Danube is also capable of simultaneously resolving multiple lifetime components within the same cell. This allows the differential quantification of lifetime changes of a given compound in the subcellular environment.

The Danube brings a new level of performance to cell analysis. By allowing the rapid measurement of subnanosecond lifetime changes across entire cell populations, it gives cell researchers a flexible and efficient new tool for the study of the subcellular environment.

Form factor of a 1-laser, 4-detector Danube. Sheath and waste tanks housed separately.


Specifications

Excitation Optics

Standard laser:

  • 488 nm (≤ 200 mW)
  • 405 nm (≤ 300 mW)
  • 640 nm (≤ 150 mW)

Custom laser options (powers vary 50 – 300 mW):

  • 375, 395, 420, 445, 460, 473, 505, 515, 633, 660, 685, 785, 850 nm
All sources pulsed at repetition rates 10 – 100 MHz  

Emission Optics

Standard channels:

  • FSC: 2 – 10°
  • SSC: 90°, 1.2 NA
  • 488 laser: FL3 (530/30), FL4 (580/30)
  • 405 laser: FL1 (430/30), FL2 (470/30)
  • 640 laser: FL5 (660/30), FL6 (710/LP)

Custom channel bandpass selections available for each custom laser option

Fluidics

Hydrostatic sheath pressure injection:

  • 8-L sheath capacity, pressure up to 30 psig

Hydrostatic sample pressure injection:

  • Injection speed: 1 – 100 µL/min

Signal Processing

Digital waveform sampling:

  • up to 1.5 GHz bandwidth
  • up to 10-bit resolution (raw data)
  • up to 2.5 GS/s per channel

Offline signal analysis:

  • multiexponential lifetime fit

Performance 

Fluorescence lifetime:

  • down to 4-ns interaction time
  • 500-ps lifetime resolution
  • multiexponential decay

Sensitivity (488-nm excitation, 530/30-nm channel):

  • FITC ≤ 1000 MESF (typ.)
  • 5/6 Spherotech Rainbow bead peaks
  • CV 6% (typ.)

Throughput:

  • 5,000 events/s (typ.)

Installation Requirements 

Dimensions:

  • 24” x 36” x 10” (W x L x H) (separate sheath and waste tanks)

Weight:

  • 160 lbs. (1-laser, 4-detector system)

Environmental:

  • 15°–30°C, 60% RH

Power:

  • North America: 120 VAC, 50/60 Hz, 5A
  • Japan: 100 VAC, 50/60 Hz, 5A
  • Rest of world: 230 VAC, 50/60 Hz, 3A


KRCDS.Danube.1v1             
† For Research Use Only. Not for diagnostic use. Specifications subject to change without notice.