Achieving more with less.

Archive of CYTO2025 commercial tutorial

Title Achieving more with less. The value of the hybrid photodetector (Hypereon) in flow cytometry
Date June 2, 2025 • 12:15 - 1:15 PM MDT
Room Mile High 2C & 3C

 

 

Measurement of four mixed samples

Measurement of four mixed samples with one laser and one gain operation

4 mixed samples

  • APC Single stain (30 ng)
  • AF647 Singl stain (30 ng)
  • APC+AF647 (30 ng:15 ng)
  • APC+AF647 (15 ng:30 ng)

 

Compensation Bead;  BD™ Anti-Rat Ig, κ/Negative Control (BSA) Compensation Plus (7.5 µm) Particles Set [Cat. No. 560499]

Fluorescence Antibody Reagent; APC anti-mouse CD62L Antibody [Cat. No. 161217]

       Alexa Fluor® 647 anti-mouse CD197 (CCR7) Antibody [Cat. No. 120111]

Buffer; BD PharmingenTM Stain Buffer(FBS) [Cat. No. 554656]

Separation accuracy varies depending on the channel selected

Hypereon

Acceptable

APC:657

AF647:670

Optimal

APC:657

AF647:684

Suboptimal

APC:670

AF647:684

Optimal

APC:657

AF647:697

PMT

Unacceptable

APC:670

AF647:695

Unacceptable

APC:644

AF647:695

Unacceptable

APC:670

AF647:718

Measurement of ten mixed samples

Measurement of ten mixed samples with one laser and one gain operation

10 mixed samples

  • APC Single stain 160ng
  • APC Single stain 40ng
  • APC Single stain 3.2 ng
  • AF647 Single stain 160ng
  • AF647 Single stain 40ng
  • AF647 Single stain 3.2 ng

  • APC + AF647 160ng : 160ng
  • APC + AF647 40ng : 20 ng
  • APC + AF647 20ng : 40ng
  • APC + AF647 3.2ng : 3.2ng

 

Compensation Bead;  BD™ Anti-Rat Ig, κ/Negative Control (BSA) Compensation Plus (7.5 µm) Particles Set [Cat. No. 560499]

Fluorescence Antibody Reagent; APC anti-mouse CD62L Antibody [Cat. No. 161217]

       Alexa Fluor® 647 anti-mouse CD197 (CCR7) Antibody [Cat. No. 120111]

Buffer; BD PharmingenTM Stain Buffer(FBS) [Cat. No. 554656]

Hypereon for EV measurement

These are measurement results from the Ota Laboratory at the University of Tokyo.

These measurements were taken using a photocathode different from that used in normal Hypereon measurements.

Optics for comparison of Hypereon and PMT

Raw data from Hypereon, PMT and ground truth

Hypereon enables corrct detection of <10 photons

  1. Detect events and count photons using ground truth
  2. Judge detection by PMT and HYPEREON for each event
  3. Calculate accuracy as mAP (mean Average Precision)
  4. Plot mAP vs. photon count for each detector

Hypereon improves LoD(Limit of Detection) and dynamic range

Scattering

Fluorescence

Measurement window width 1 µs, Laser wavelength 405nm, Laser intensity 120 mW.

Potential application: virus and EVs detection

Fluorescence measurement

Detection window width : 1 us

Estimation of detectable Particle diameter

Detection window width : 1 us

Measurement window width 1 µs, Laser wavelength 405nm, Laser intensity 120 mW.

Data provision and cooperation

Yuichiro Iwamoto 

Postdoctoral Researcher, Ota Lab, The University of Tokyo

Specialized in high-throughput, single-particle analysis by integrating optics, fluidics, and data science.

The CYTO2024 seminar archive | held in Edinburgh

This is an archived video of the presentation given at CYTO2024.

As this presentation was given in 2024, some of the information may not reflect the latest information. Please note that this is for reference purposes only.

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