sp6800 spectral analyzer · 2017-08-02 · sp6800 spectral analyzer the sony sp6800 spectral...

Sony Biotechnology Inc.

SP6800Spectral Analyzer

SP6800 Spectral Analyzer

The Sony SP6800 Spectral Analyzer uses spectral technology to optimize sensitivity and enhance dim signal detection by collecting photons from 420nm to 800nm. Spectral technology also simplifies multicolor panel design, by eliminating band-pass filters and conventional compensation matrices while delivering better data and simplifying visualization for the study of heterogeneous populations.

Novel global standardization mode automatically sets the system to a master specification with a single click. This capability eliminates instrument variability from day to day and across multiple instruments for greater reliability.

Advanced electronics and patented optical technologies bring simplicity to Spectral Analysis workflows. Sony’s patented FlowpointTM core stability and tracking system and automated QC ensure the highest resolution possible of target populations.

The system features easy to use software that automates alignment and laser delay with set up wizards and simplified voltage settings. Each system includes FCS ExpressTM software in addition to Sony analysis packages to offer the highest flexibility in analysis.

• Spectral analysis technology optimizes sensitivity while simplifying application design and workflow.

• Enhances dim signal detection for better visualization of rare populations, fluorescent proteins, and fluorochromes excited by multiple lasers.

• Novel global standardization mode automatically sets the system to a master specification with a single click.

• Easy to use software features include automated alignment and laser delay via set up wizards, easy acquisition, and flexible analysis using both Sony and FCS Express software included with every system.

3

The SP6800 spectral flow analyzer improves sensitivity and simplifies application design, workflow, and analysis over conventional flow cytometers. This is achieved using spectral analysis technology, advanced electronics, and patented optical technologies. These capabilities, unique to Sony Biotechnology systems, allow experienced and novice flow cytometrists to achieve greater flexibility for panel design and more accurate visualization of results.

SP6800 System Overview

Microfluidics flow cell chip maximizes signal with auto positioning to guarantee high sensitivity. Made of durable plastic with an embedded quartz cuvette, the chip is easy to replace when needed.

The 405nm, 488nm and 638nm excitation lasers are positioned to reduce fluorescent noise. They enable the system to support 16 or more fluorescent parameters.

Emitted light is directed through through a 32 Channel PMT that produces 66 data points of signal detection to analyze emitted photons from 420nm to 800 nm to ensure accurate visualization.

A unique prism collection systemDelivers light through 10 consecutive prisms allowing optimal signal separation while minimizing light loss.

The Flowpoint detection system precisely tracks the core stream shape and position in the flow cell as well as the cross sectional position of each passing particle to provide highly reliable measurements. This patented technology visualizes core stability and enables the highest resolution.

Scatter analysisForward and Side Scatter parameters to allow relative size and complexity measurements.

All events

FSC_A

SSC

_A

20

4

3

2

1

40 60x1,000

x1,0

00

4

The SP6800 software is easy to learn and use. It guides researchers from set-up to panel design, acquisition, analysis, and shutdown.

User preferences allow users and administrators set up options for overall instrument operation and experiment set up to facilitate use and unattended operation procedures.

Software

Standardization mode sets the system to a master specification to eliminate variability in a single instrument or among instruments located across sites. This unique capability allows experiments performed on any Sony cell analyzer to produce highly reliable, accurate, and reproducible results. This function also eliminates setup subjectivity for collaborative or long term studies with different operators or experience levels across sites.

Standardization

At start up Align Check and Performance QC wizards check instrument calibrations, using beads to ensure the instrument is operating optimally. On screen instructions guide the user through procedures, then display progress and report results. The performance report displays MESF, Q, and B values to describe real-time fluorescence detection performance. If desired, Align Check and Performance QC reports can be displayed in historical context.

System Start Up

When engaged, standardization mode sets the SP6800 to predetermined global settings that eliminate instrument variability.

5

On shutdown, the SP6800 software guides the user though pre –shutdown cleaning and shuts down the instrument automati-cally. Software wizards are also available to guide users through Bleach Cleaning and Rinse procedures.

All acquisition functions, including instru-ment settings are controlled from the Acquisition Window. Worksheet tools let users choose how the data is displayed- (such as plot types), and customize for their analysis needs. Plots and statistics provide real-time information during acquisition. To increase sample acquisition to the cuvette, a variable booster lets users set acquisition speed from low (33ul/min) normal or high (250ul/min) offering flexibility.

Acquisition Functions and Analysis System Shutdown

Experiments can be created using a template, an existing experiment, a single stained, or multicolor assay, in the Create Experiment window. Users can point and click to choose (or edit) an existing ex-periment and can easily select templates for wells or plates when creating a new experiment. A setup Assay Wizard guides users through the creation of a single-stained or multicolor assay simplifying experiment creation.

The Spectral Library lets users create a personal reagent library that simplifies ex-periment creation and saves time. An Acquisition Wizard assists the users with step by step instructions to acquire and analyze single positive controls for your spectral library. Once acquired the spectral reference for that reagent including the spectral index are available for future use. Information from the spectral library is avail-able to users with a simple click improving accuracy and streamline panel design.

Spectral Library Experiment Creation

6

Spectral Analyzers from Sony include FCS Express, from De Novo Software. FCS Express offers a range of new analysis tools from live gating to batchanalysis. Native support for Sony Spectral data files to enable sophisticateddata transformations and visualizations such as spectral overlays, tSNE,Spade, and heat maps.

FCS Express from De Novo Software

105

106

FOX

P3

: PE-

Daz

zle5

94 -

Are

a

CD4 : BV421 - Area

104

103

-102

102

-103

103 104 105 106

Inte

nsi

ty

Height (Wavelength)

FOXP3 +

FOXP3-

420.0 515.0 610.0 705.0 800.0

107

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105

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Inte

nsi

ty

Height (Wavelength)

FOXP3 +

FOXP3-

420.0 515.0 610.0 705.0 800.0

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105

FOX

P3

: PE-

Daz

zle5

94

- A

rea

CD4 : BV421 - Area

104

103

102

102

-102

103 104 105 106

SSC

- A

rea

(x 1

00

0)

CD45 : BV711 - Area

Lymphs

262.1

196.6

131

65.5

102-0.1

-103 103 104 105 106

Inte

nsi

ty

Height (Wavelength)

FOXP3 +

FOXP3-

420.0 515.0 610.0 705.0 800.0

107

106

105

104

103

102

101

Dot plots and spectral plots can be set up with live gating. Live gating allows users to find positive and negative populations quickly and easily for added assurance about target populations.

Live Gating

Flexible data analysis can include integrated spreadsheets, custom calculations, charting and regression analysis.

Batch analysis lets you process any number of samples with one click. To support presentation, data can be exported directly to PowerPoint, PDF, and Excel.

Flexible Analysis Batch and Report

7

Spectral Analysis Technology

Spectral analysis technology is the foundation of the SP6800 system. Spectral flow cytometry streamlines workflow and yields better data by keeping all the light collected. In conventional systems, overlapping fluorescence is subtracted using color compensation, so less light is collected. Instead, spectral flow cytometers sum the fluorescence together and then use unmixing to mathematically separate the colors. This powerful capability also simplifies workflow including panel design, and improves visualization of autofluorescence.

A unique prism collection system delivers emitted light to a 32 channel PMT. This produces 66 data points of signal detection for fluorescence and bright auto fluorescence to achieve accurate visualizations of fluorescent populations. This lets researchers see the complete spectral fingerprintof each fluorochrome from 420nm to 800nm.

Visualization of fluorescent cell populations using the analyzer’s unique prism collection system.

A powerful capability of spectral technology is Unmixing. This allows researchers to separate fluorophores into pure signals that measure the quantity of each fluorophore at each pixel to more accurately measure data for analysis.

Spectral unmixing separates each spectral fingerprint to better visualize each fluorochrome marker. Unlike conventional filtering where overlapping signals are lost, spectral unmixing captures the photons emitted from 420nm to 800nm. In doing so it enhances dim signal detection for better visualization of rare populations, fluorescent proteins and fluorochromes excited by multiple lasers.

This also lets researchers separate the spectra of fluorophores masked by autofluorescence by extracting it from the signal and creating it into a unique fluorescent parameter. With a clear signal for each color channel unaffected by overlapping signals (spillover) and autofluorencence, spectral analysis yields better, unbiased data for analysis.

Spectral UnmixingLD

-48

8_A

106

104

103

102

101

100

105

E

Wavelengths500 600 700 800

Spectral analysis reduces false-positives and delivers more accurate analysis over conventional flow cytometry. Mouse splenocytes were stained with CD4 APC and anti-IL-2 PE. A. In this conventional density plot it is unclear if the light-blue region is a dim PE, weak double positive, or non-specific binding. B. Using Spectral analysis the spectral data of each region is compared against the Spectral Library to unmix the sample. This reveals the light blue region is high auto-fluorescence. Representative data collected on SP6800.

CD

4_A

PC

_ALD

-48

8_A

105

104

103

102

101

-101

106

104

103

102

101

100

105

E: 95.81%

G: 2.18%

F: 1.28%

CD

4_A

PC

_A

IL2_PE_A

105

104

103

102

101

-101

-102100102

E: 95.65%

G: 8.63%

F: 1.42%

103 104 105

E

Wavelengths500 600 700 800

Wavelengths Wavelengths

LD-4

88

_A

106

104

103

102

101

100

105

G

500 600 700 800

LD-4

88

_A

106

104

103

102

101

100

105

F

500 600 700 800

102 104 105-102100

IL2_PE_A103

A B

Unstained cells

HighAuto-fluorescence

PE Positive cells

Unmixing

=Spectral Unmixing separates each spectral fingerprint for complete and optimal visualization of fluorochromes.

8

H-Q2: 3.50%H-Q1: 88.02% H-Q1: 91.66%

H-Q4: 0.15%H-Q4: 0.07%

H-Q3: 8.33%H-Q3: 8.24%

All events105

104

103

102

-101

-102

-102 102

0GFP_A

Ale

xa F

luo

r 70

0_A

103 104 105

H-Q2: 0.04%

All events105

104

103

102

-101

-102

-102 102

0GFP_A

Ale

xa F

luo

r 70

0_A

103 104 105

H-Q2: 3.50%H-Q1: 88.02% H-Q1: 91.66%

H-Q4: 0.15%H-Q4: 0.07%

H-Q3: 8.33%H-Q3: 8.24%

All events105

104

103

102

-101

-102

-102 102

0GFP_A

Ale

xa F

luo

r 70

0_A

103 104 105

H-Q2: 0.04%

All events105

104

103

102

-101

-102

-102 102

0GFP_A

Ale

xa F

luo

r 70

0_A

103 104 105

Y: 0.27%

S: 0.30%

R: 95.62%

F4/80_PE_A

Wavelength

S

LD40

5/63

8_A

420

106

105

104

103

102

-101

-102

550 680 800

O106

105

104

103

102

-101

-102

-102 -101 102 103 104 105 106

B22

0_B

V65

0_A

T: 0.39%

Wavelength

V

LD40

5/63

8_A

420

106

105

104

103

102

-102

550 680 800

Wavelength

Y

LD40

5/63

8_A

420

106

105

104

103

102

-102

550 680 800

GFP Pos: 0.90%

GFP neg: 98.74%

All events106

-103

GFP_A

SSC

_A

105

104

103

102

101

100

-102 103 104 105

GFP Pos: 0.21%

GFP neg: 99.66%

All events106

-103

GFP_A

SSC

_A

105

104

103

102

101

100

-102 103 104 105

GFP Pos: 0.90%

GFP neg: 98.74%

All events106

-103

GFP_A

SSC

_A

105

104

103

102

101

100

-102 103 104 105

GFP Pos: 0.21%

GFP neg: 99.66%

All events106

-103

GFP_A

SSC

_A

105

104

103

102

101

100

-102 103 104 105

In conventional flow cytometry cellular auto-fluorescence produced by pyridine (NAD/NADH), flavin (FMN, FAD), and other intracellular oxidative reactions can cause fluorescent signal contamination of other fluorescent markers. Other common sources of auto-fluorescence include cell fixation and permeabilization. Using spectral technology, auto-fluorescent spectral fingerprints can be subtracted to allow researchers to see the true fluorescent population.

Unstained mouse splenocytes were analyzed with the SP6800 revealing three distinct auto-fluorescent populations. Using the spectral fingerprints obtained in analysis, the appropriate auto-fluorescence can be removed, increasing the precision and quality of results.

Auto-fluorescence can result in the appearance of additional cell populations leading to erroneous data interpretation. A. T cells expressing GFP and stained with an antibody conjugated to Alexa Fluor® 700 were analyzed with the SP6800. A double positive population is present in the uncorrected density plot. B. This double positive population disappears when the auto-fluorescent spectral fingerprint is subtracted. C. Liposomes from cells expressing GFP were analyzed. D. This uncovered a small positive GFP population after auto-fluorescence was subtracted.

Subtracting Auto-Fluorescence Improves Visualization

A. With Auto-fluorescence

C. With Auto-fluorescence

B. Without Auto-fluorescence

Activated T Cells with GFP and Alexa Fluor 700

Liposomes with GFP

D. Without Auto-fluorescence

A correction system adjusts the offset and sensitivity of each channel of the 32 channel PMT to ensure a uniform and accurate measurement of fluorescent emission from 420nm to 800nm. The corrective system brings a standardization to all 32 PMTs ensuring the user is getting the most reliable data with the ease of adjusting only one voltage. This saves time over conventional flow cytometry operation where users must calibrate each individual PMT.

Uniform measurement of Fluorescent Emissions

Pre and Post Correction Profile. Each graph illustrates pre and post correction in the SP6800 32 channel PMT. The correction improves accuracy of spectral visualization.

Spectral emission of PE Cy5 pre and post correction. Corrections support accurate unmixing of closely overlapping fluorescence spectra.

Post-correction120

100

80

60

40

20

01 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31

Pre-correction120

100

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40

20

01 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31

Pre correction Post correction

PE-Cy5 PE-Cy5

All events

Channel

LD4

88

_H

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1 10 20 32

All events

Channel

LD4

88

_H

106

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101

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1 10 20 32

Correction

9

500 600Wavelength (nm)

700 800

Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8

• Alexa Fluor® 488; 488nm • Alexa Fluor 532; 488nm • PE; 488nm • PE DazzleTM 594; 488nm • PE Alexa Fluor 610; 488nm • PE Cy5TM; 488nm • PerCP Cy5.5; 488nm • PE Cy7; 488nm

• BrilliantTM Blue 515; 488nm • Alexa Fluor 514; 488nm • PE Texas Red®; 488nm • PerCP eFluor 710; 488nm • PE Cy5.5; 488nm • PE Vio®770; 488nm

• FITC; 488nm • PerCP; 488nm

• PE Alexa Fluor 700; 488nmLaser

• 488nm

wavelength (nm)500 600

wavelength (nm)400 600500


• Brilliant VioletTM 421; 405nm • Pacific BlueTM; 405nm • Brilliant Violet 510TM; 405nm • Brilliant Violet 570TM; 405nm • Brilliant Violet 605TM; 405nm • Brilliant Violet 650TM; 405nm • Brilliant Violet 711TM; 405nm • Brilliant Violet 785TM; 405nm

• Alexa Fluor 405; 405nm • HorizonTM V450; 405nm • AmCyan; 405nm • Pacific OrangeTM; 405nm • Qdot® 605; 405nm • Qdot 655; 405nm • Qdot 705; 405nm • Qdot 800; 405nm

• Horizon V500; 405nm • eVolveTM 605; 405nm • eFluor 660; 405nm • Alexa Fluor 700; 638nm • APC Cy7; 638nm

• Krome OrangeTM; 405nm • Alexa Fluor 647; 638nm • APC Cy5.5; 638nm • APC Alexa 750; 638nm

• APC; 638nm • APC eFluor 780; 638nm

• Cy5; 638nm

400 500 600

Wavelength (nm)

700 800

Laser

• 405nm

• 638nm

Application data dye options and combinations. Select one from each group.

Fluorochrome Panel Guide

10

500 600Wavelength (nm)

700 800


• Alexa Fluor® 488; 488nm • Alexa Fluor 532; 488nm • PE; 488nm • PE DazzleTM 594; 488nm • PE Alexa Fluor 610; 488nm • PE Cy5TM; 488nm • PerCP Cy5.5; 488nm • PE Cy7; 488nm

• BrilliantTM Blue 515; 488nm • Alexa Fluor 514; 488nm • PE Texas Red®; 488nm • PerCP eFluor 710; 488nm • PE Cy5.5; 488nm • PE Vio®770; 488nm

• FITC; 488nm • PerCP; 488nm

• PE Alexa Fluor 700; 488nm

700 800

700 800


• Brilliant VioletTM 421; 405nm • Pacific BlueTM; 405nm • Brilliant Violet 510TM; 405nm • Brilliant Violet 570TM; 405nm • Brilliant Violet 605TM; 405nm • Brilliant Violet 650TM; 405nm • Brilliant Violet 711TM; 405nm • Brilliant Violet 785TM; 405nm

• Alexa Fluor 405; 405nm • HorizonTM V450; 405nm • AmCyan; 405nm • Pacific OrangeTM; 405nm • Qdot® 605; 405nm • Qdot 655; 405nm • Qdot 705; 405nm • Qdot 800; 405nm

• Horizon V500; 405nm • eVolveTM 605; 405nm • eFluor 660; 405nm • Alexa Fluor 700; 638nm • APC Cy7; 638nm

• Krome OrangeTM; 405nm • Alexa Fluor 647; 638nm • APC Cy5.5; 638nm • APC Alexa 750; 638nm

• APC; 638nm • APC eFluor 780; 638nm

• Cy5; 638nm

400 500 600

Wavelength (nm)

700 800

11

WLSM

Granulocytes

Lymphocytes

Monocytes

All events

x1,0

00

SSC

_A

CD45_PE-Cy7_A

5

4

3

2

1

-102-10

210

210

310

410

5

WLSM

CD24-CD13

Granulocytes

CD

13_P

erC

P e

flo

ur7

10_A

CD24_PE-AF610_A

104

-102

10110

210

310

410

5

103

102

-101

-102

WLSM

CD35-CD3

Natural Killer Cells

CD35-CD3 12.27%

Lymphocytes

CD

35_B

V4

21_A

CD3_Alexa Fluor 458_A

106

-102

100

102

103

105

104

103

-102

WLSM

CD35-CD3

CD

16_B

V60

5_A

CD3_Alexa Fluor 458_A-10

2-10

110

210

6

104

103

-102

103

104

105

CD16-CD3: 10.73%

WLSM

Lymphocytes

CD

16_B

V60

5_A


2-10

010

2

104

103

-102

103

CD11c-CD14-

WLSM

CD45+

CD

14_B

V78

5_A

CD11c_BV711_A-10

210

2

104

103

-102

-103

-103

106

105

104

CD14 positive

WLSM

Monocytes

CD

14_B

V78

5_A

CD11c_BV711_A-10

210

3

104

103

102

-101

-102

-103

105

104

Cytotoxic T Cells

WLSM

Lymphocytes

CD

8_P

E_A


210

3

104

103

102

-102

-10010

2

Naive Cytotoxic T Cells

E�ector Cytotoxic T Cells E�ector Cytotoxic T Cells

WLSM

Cytotoxic T Cells

CD

45R

A_B

V51

0_A

CD35_BV421_A-10

210

3

104

103

102

-101

-102

-103

102

101

Helper T Cells

WLSM

Lymphocytes

CD

4_P

EDaz

zle

594

_A


210

3

103

-101

-102

-10010

2

Naive Helper T Cells

WLSM

Helper T Cells

CD

45R

A_B

V51

0_A

CD35_BV421_A-10

210

3

103

102

-102

-103

-101

-10010

2

CD13-CD14

WLSM

CD4

CD

14_B

V78

5_A

CD13_PerCP eflour710_A-10

210

3

103

102

-101

-102

104

CD3-CD33

WLSM

All events

CD

14_B

V78

5_A

CD11c_BV711_A-10

210

3

104

103

102

-102

-103

105

104

PLASMA B Cells

WLSM

CD19-CD20

CD

35_B

V4

21_A

HLA-DR_PacificBlue_A-10

210

3

104

103

102

-102

-101

104

104

104

CD19c-CD20

WLSM

CD11c-CD14-

CD

23_P

E-C

y5.5

_A

CD19_PE-Cy5_A-10

210

3

104

103

-102

104

Memory B Cells

Naive B Cells

WLSM

PLASMA B Cells

CD

11b

_BV

650

_A

CD45RA_BV510_A10

210

3

104

105

103

102

-102

-101

-103

101

100

104

105

106

WLSM

Granulocytes

Lymphocytes

Monocytes

All events

x1,0

00

SSC

_A

CD45_PE-Cy7_A

5

4

3

2

1

-102-10

210

210

310

410

5

WLSM

CD24-CD13

Granulocytes

CD

13_P

erC

P e

flo

ur7

10_A

CD24_PE-AF610_A

104

-102

10110

210

310

410

5

103

102

-101

-102

WLSM

CD35-CD3


CD35-CD3 12.27%

Lymphocytes

CD

35_B

V4

21_A


106

-102

100

102

103

105

104

103

-102

WLSM

CD35-CD3

CD

16_B

V60

5_A


2-10

110

210

6

104

103

-102

103

104

105

CD16-CD3: 10.73%

WLSM

Lymphocytes

CD

16_B

V60

5_A


2-10

010

2

104

103

-102

103

CD11c-CD14-

WLSM

CD45+

CD

14_B

V78

5_A

CD11c_BV711_A-10

210

2

104

103

-102

-103

-103

106

105

104

CD14 positive

WLSM

Monocytes

CD

14_B

V78

5_A

CD11c_BV711_A-10

210

3

104

103

102

-101

-102

-103

105

104

Cytotoxic T Cells

WLSM

Lymphocytes

CD

8_P

E_A


210

3

104

103

102

-102

-10010

2



WLSM

Cytotoxic T Cells

CD

45R

A_B

V51

0_A

CD35_BV421_A-10

210

3

104

103

102

-101

-102

-103

102

101

Helper T Cells

WLSM

Lymphocytes

CD

4_P

EDaz

zle

594

_A


210

3

103

-101

-102

-10010

2


WLSM

Helper T Cells

CD

45R

A_B

V51

0_A

CD35_BV421_A-10

210

3

103

102

-102

-103

-101

-10010

2

CD13-CD14

WLSM

CD4

CD

14_B

V78

5_A


210

3

103

102

-101

-102

104

CD3-CD33

WLSM

All events

CD

14_B

V78

5_A

CD11c_BV711_A-10

210

3

104

103

102

-102

-103

105

104

PLASMA B Cells

WLSM

CD19-CD20

CD

35_B

V4

21_A


210

3

104

103

102

-102

-101

104

104

104

CD19c-CD20

WLSM

CD11c-CD14-

CD

23_P

E-C

y5.5

_A

CD19_PE-Cy5_A-10

210

3

104

103

-102

104

Memory B Cells

Naive B Cells

WLSM

PLASMA B Cells

CD

11b

_BV

650

_A

CD45RA_BV510_A10

210

3

104

105

103

102

-102

-101

-103

101

100

104

105

106

WLSM

Granulocytes

Lymphocytes

Monocytes

All events

x1,0

00

SSC

_A

CD45_PE-Cy7_A

5

4

3

2

1

-102-10

210

210

310

410

5

WLSM

CD24-CD13

Granulocytes

CD

13_P

erC

P e

flo

ur7

10_A

CD24_PE-AF610_A

104

-102

10110

210

310

410

5

103

102

-101

-102

WLSM

CD35-CD3


CD35-CD3 12.27%

Lymphocytes

CD

35_B

V4

21_A


106

-102

100

102

103

105

104

103

-102

WLSM

CD35-CD3

CD

16_B

V60

5_A


2-10

110

210

6

104

103

-102

103

104

105

CD16-CD3: 10.73%

WLSM

Lymphocytes

CD

16_B

V60

5_A


2-10

010

2

104

103

-102

103

CD11c-CD14-

WLSM

CD45+

CD

14_B

V78

5_A

CD11c_BV711_A-10

210

2

104

103

-102

-103

-103

106

105

104

CD14 positive

WLSM

Monocytes

CD

14_B

V78

5_A

CD11c_BV711_A-10

210

3

104

103

102

-101

-102

-103

105

104

Cytotoxic T Cells

WLSM

Lymphocytes

CD

8_P

E_A


210

3

104

103

102

-102

-10010

2



WLSM

Cytotoxic T Cells

CD

45R

A_B

V51

0_A

CD35_BV421_A-10

210

3

104

103

102

-101

-102

-103

102

101

Helper T Cells

WLSM

Lymphocytes

CD

4_P

EDaz

zle

594

_A


210

3

103

-101

-102

-10010

2


WLSM

Helper T Cells

CD

45R

A_B

V51

0_A

CD35_BV421_A-10

210

3

103

102

-102

-103

-101

-10010

2

CD13-CD14

WLSM

CD4

CD

14_B

V78

5_A


210

3

103

102

-101

-102

104

CD3-CD33

WLSM

All events

CD

14_B

V78

5_A

CD11c_BV711_A-10

210

3

104

103

102

-102

-103

105

104

PLASMA B Cells

WLSM

CD19-CD20

CD

35_B

V4

21_A


210

3

104

103

102

-102

-101

104

104

104

CD19c-CD20

WLSM

CD11c-CD14-

CD

23_P

E-C

y5.5

_A

CD19_PE-Cy5_A-10

210

3

104

103

-102

104

Memory B Cells

Naive B Cells

WLSM

PLASMA B Cells

CD

11b

_BV

650

_A

CD45RA_BV510_A10

210

3

104

105

103

102

-102

-101

-103

101

100

104

105

106

Application data using 2 lasers

16 color panel on Human Peripheral Blood

488nm Laser 405nm Laser

Examples of the resolution of several important cell populations using only two lasers with the SP6800. Clear population resolution is obtained with highly overlapping fluorochomes such as PE-Cy5/PE-Cy5.5 (G) and Pacific Blue/BV421 (H).

A

F

J

B

G

K

C

H

L

D

I

E

HLA-DR Pacific Blue

CD38 BV421

CD45RA BV510

CD33 BV570

CD16 BV605

CD11b BV650

CD11c BV711

CD14 BV785

CD3 Alexa Fluor 488

CD8 PE

CD4 PE Dazzle 594

CD24 PE Alexa Fluor 610

CD19 PE Cy5

CD20 PE Cy5.5

CD13 PerCP efluor® 710

CD45 PE Cy7

2 Lasers, 488nm Laser


400 800700600

Wavelength (nm)

500

400 800700600

Wavelength (nm)

500



400 800700600

Wavelength (nm)

500

400 800700600

Wavelength (nm)

500

Sample Data

12

WLSM

Granulocytes

CD13+CD24+ Grans

Lymphocytes

Monocytes

All events

x1,0

00

SSC

_A

CD

13_P

erC

P e

flo

ur7

10_A

CD45_PE-Cy7_A CD24_PE-AF620_A

4

2

-102

-101

102

103

104

105

106

WLSM

Plasma B Cells: 99.57%

CD19+CD20+

CD

38_A

PC

_A


2-10

110

210

310

410

5

105

104

103

102

-101

-102

-103

106

WLSM

Helper T Cells: 50.38%

Lymphocytes

CD

4_P

EDaz

zle

594

_A


2-10

110

210

3

101

-100

-101

WLSM

Helper Naive T Cells

Lymphocytes

CD

45R

A_A

PC-

Cy7

_A

CD38_APC_A-10

210

210

3

103

102

104

-102

WLSM

Cytotoxic T Cells

Lymphocytes

CD

8_P

E_A


2-10

110

210

310

4

104

103

102

-102

WLSM

Cytotoxic Naive T Cells

Cytotoxic E�ector T Cells

Helper E�ector T Cells

WLSM

CD5+CD24+ B Cells

Lymphocytes

CD

24_P

E-A

F610

_A

CD5_BV421_A-10

210

310

5

104

103

105

106

-102

-103

-103

104

WLSM

CD13+CD33+ Monocytes

CD

33_B

V57

0_A


210

3

102

101

103

-102

-102

Cytotoxic T Cells

CD

45R

A_A

PC-

Cy7

_A

CD38_APC_A-10

210

210

310

4

104

103

102

-102

WLSM

CD16 positive

Monocytes

CD

16_B

V60

5_A


210

210

310

4

Monocytes

WLSM

CD24+CD38- Granulocytes

CD

38_A

PC

_A

CD24_PE-AF610_A-10

210

3

104

103

106

105

-102

-103

-101

-103

102

104

104

103

-102

-100

WLSM

Granulocytes

103

-102-10

110

210

310

4

102

-101

-102

-103

-103

NK Cells

CD

16_B

V60

5_A


WLSM

Lymphocytes

104

-102-10

110

210

3

103

102

-102

CD19+CD20+C

D20

_PE-

Cy5

.5_A

CD19_PE-Cy5_A

WLSM

CD45+

104

-103

-102

103

104

105

103

-102

-103

Granulocytes

Application data using 3 lasers

16 color stained sample of Human Peripheral Blood

488nm Laser

638nm Laser

405nm Laser

Examples of the detection of several important cell populations using sixteen fluorochromes excited by three lasers. Unlike conventional flow cytometers that can detect up to five off the blue, in this example we demonstrate that with the SP6800 eight fluorochromes can be excited off the blue with clear sample resolution. Even highly overlapping fluorchromes such as PE-Cy5 and PE-Cy5.5 can be resolved.

CD3 Alexa Fluor 488

CD8 PE

CD4 PE Dazzle 594

CD24 PE Alexa Fluor 610

CD19 PE Cy5

CD20 PE Cy5.5

CD13 PerCP efluor 710

CD45 PE Cy7

HLA-DR Pacific Blue

CD5 BV421

CD56 BV510

CD33 BV570

CD16 BV605

CD38 APC

CD11c Alexa Fluor 700

CD45RA APC-Cy7


400 800700600

Wavelength (nm)

Wavelength (nm)

Wavelength (nm)

500

400 800700600500

400 800700600500




400 800700600

Wavelength (nm)

Wavelength (nm)

Wavelength (nm)

500

400 800700600500

400 800700600500




400 800700600

Wavelength (nm)

Wavelength (nm)

Wavelength (nm)

500

400 800700600500

400 800700600500



13

Spectral Analysis allows multi-laser excited fluorochromes to be run without using a complicated compensation matrix.

Application Data: Brilliant Violet Dyes

Wavelength (nm)

400 800700600500

Example of Human Peripheral Blood Leukocytes was analyzed on the SP6800 and a BD LSRFortessa conventional flow cytometer. Spectral analysis was able to remove the autofluorescenece and use unmixing to separate each fluorochrome which resulted in clarity and resolution of each population in these density plots.

Sample data from Human PBMCs stained with seven markers conjugated to Brilliant Violet dyes offered by Sony Biotechnology. D. All populations were clearly resolved including fluorochomes with significant spectral overlap such as BV605 and BV650 (D).

12-color Staining of Human Peripheral Blood Leukocytes

BV421

BV510

BV570

BV605

BV650

BV711

BV785

BA C D E

CD45+

SSC

_A

CD

8_B

V57

0_A

CD45_BV711_H CD3_BV421_A

H

x1,0

00

10

8

6

4

2

102 104 105103

CD45+

102

104

103

102

-101

-102

-102 103 104 105

CD

4_B

V51

0_A

CD3_BV421_A

CD45+

102

103

102

-101

-102

-102 103 104 105C

D16

/56_

BV

605_

ACD19_BV650_A

CD45+

102

103

102

-102

-102 -101 103

CD

3_B

V4

21_A

CD14_BV785_A

CD45+

102

105

104

103

-102

102

-102-100 103

CD45+

SSC

_A

CD

8_B

V57

0_A


H

x1,0

00

10

8

6

4

2

102 104 105103

CD45+

102

104

103

102

-101

-102

-102 103 104 105

CD

4_B

V51

0_A

CD3_BV421_A

CD45+

102

103

102

-101

-102

-102 103 104 105C

D16

/56_

BV

605_

A

CD19_BV650_A

CD45+

102

103

102

-102

-102 -101 103

CD

3_B

V4

21_A

CD14_BV785_A

CD45+

102

105

104

103

-102

102

-102-100 103

CD45+

SSC

_A

CD

8_B

V57

0_A


H

x1,0

00

10

8

6

4

2

102 104 105103

CD45+

102

104

103

102

-101

-102

-102 103 104 105

CD

4_B

V51

0_A

CD3_BV421_A

CD45+

102

103

102

-101

-102

-102 103 104 105C

D16

/56_

BV

605_

A

CD19_BV650_A

CD45+

102

103

102

-102

-102 -101 103

CD

3_B

V4

21_A

CD14_BV785_A

CD45+

102

105

104

103

-102

102

-102-100 103

CD45+

SSC

_A

CD

8_B

V57

0_A


H

x1,0

00

10

8

6

4

2

102 104 105103

CD45+

102

104

103

102

-101

-102

-102 103 104 105

CD

4_B

V51

0_A

CD3_BV421_A

CD45+

102

103

102

-101

-102

-102 103 104 105C

D16

/56_

BV

605_

ACD19_BV650_A

CD45+

102

103

102

-102

-102 -101 103

CD

3_B

V4

21_A

CD14_BV785_A

CD45+

102

105

104

103

-102

102

-102-100 103

CD45+

SSC

_A

CD

8_B

V57

0_A


H

x1,0

00

10

8

6

4

2

102 104 105103

CD45+

102

104

103

102

-101

-102

-102 103 104 105

CD

4_B

V51

0_A

CD3_BV421_A

CD45+

102

103

102

-101

-102

-102 103 104 105C

D16

/56_

BV

605_

ACD19_BV650_A

CD45+

102

103

102

-102

-102 -101 103

CD

3_B

V4

21_A

CD14_BV785_A

CD45+

102

105

104

103

-102

102

-102-100 103

Wavelength

All events

LD4

88

_H

700 800600500102

103

104

105

106

Wavelength

All events

LD40

5/6

38_H

680 800550420102

103

104

105

106

CD45RA

Marker Fluorochrome 488nm Laser 405/638nm Laser

TCR-gd

CD56

CD45

CD3

CCR7

CD27

CD33

CD4

CD19

CD8

HLADR

Alexa Fluor 488

PE

PE-Dazzle594

PerCP-Cy5.5

PE-Cy7

BV421

BV510

BV570

BV605

APC

Alexa Fluor 700

APC-Cy7

Sample Data

14

Sony SP6800 Spectral Analyzer

CD4: BV605 - Area CD4: BV605 - Area CD4: BV605 - Area CD4: BV605 - Area

CD

27: B

V51

0 -

Are

a10

610

510

40

103 104 105 106-103

Conventional Flow Cytometer

103

-10

3

CD

19: A

PC

- A

rea

106

105

104

0-1

04

103 104 105 106-103

TCR

-gd

: PE

- A

rea

106

105

104

103

-10

3

103 105 106-103

CD

56: P

E-D

azzl

e594

- A

rea

106

105

104

-10

3

104103 105 106-103

103

CD4-BV605 CD4-BV605 CD4-BV605 CD4-BV605

CD

27-B

V51

010

510

410

310

2

0 103 104 105102-102

0

CD

19-A

PC

105

102

-10

2

105-102

104

103

1020 103 104

TCR

GD

-PE

105

104

103

102

-10

2

0 103 104 105-103 102

CD

56-P

E D

azzl

e10

510

410

3

102 103 104 105-102

102

-10

2

0


CD3: PE-Cy7 - Area

CD

45R

A: A

lexa

Flu

or

48

8 -

Are

a10

610

510

40

103 104 105 106-103


103

CD3: PE-Cy7 - Area

CCR

7: B

V4

21 -

Are

a10

610

510

410

3-1

03

103 104 105 106-103

CD8: Alexa Fluor 700 - Area

CD

56: P

E-D

azzl

e594

- A

rea

106

105

104

103

-10

3

0 104 105 106-104

CD3: PE-Cy7 - Area

HLA

DR

: AP

C-C

y7 -

Are

a10

610

510

4-1

03

0 104 105 106-104

103

CD3-PECy7

CD

45R

A-A

F48

810

510

410

310

2

0 103 104 105102-102

CD3-PECy7

CCR

7-B

V4

2110

510

20

-10

2

103 105-102

104

103

1020 103 104

CD8-AF700

CD

56-P

E D

azzl

e10

510

410

310

2-1

02

0 103 104 105-103

CD19-APC

HLA

-DR

-AP

CCy7

105

104

103

102 103 104 105-102

102

-10

20


CD3: PE-Cy7 - Area

CD

45R

A: A

lexa

Flu

or

48

8 -

Are

a10

610

510

40

103 104 105 106-103


103

CD3: PE-Cy7 - Area

CCR

7: B

V4

21 -

Are

a10

610

510

410

3-1

03

103 104 105 106-103

CD8: Alexa Fluor 700 - Area

CD

56: P

E-D

azzl

e594

- A

rea

106

105

104

103

-10

3

0 104 105 106-104

CD3: PE-Cy7 - Area

HLA

DR

: AP

C-C

y7 -

Are

a10

610

510

4-1

03

0 104 105 106-104

103

CD3-PECy7

CD

45R

A-A

F48

810

510

410

310

2

0 103 104 105102-102

CD3-PECy7

CCR

7-B

V4

2110

510

20

-10

2

103 105-102

104

103

1020 103 104

CD8-AF700

CD

56-P

E D

azzl

e10

510

410

310

2-1

02

0 103 104 105-103

CD19-APC

HLA

-DR

-AP

CCy7

105

104

103

102 103 104 105-102

102

-10

20


CD45: PerCP-Cy5.5 - Area

SSC

- A

rea

(x 1

0)

250

020

00

150

010

00

500

0

0 103 104 105 106-103


CD3: PE-Cy7 - Area

CD

4: B

V60

5 -

Are

a10

610

510

410

3-1

03

0

103 104 105 106-103

CD3: PE-Cy7 - Area

CD

8: A

lexa

Flu

or

700

- A

rea

106

105

104

0-1

04

0

103 104 105 106-103

CD3: PE-Cy7 - Area

CD

56: P

E-D

azzl

e594

- A

rea

106

105

104

0-1

03

0

103 104 105 106-103

103

CD45-PerCPCy5.5

SSC-

A(x

10

)30

00

250

020

00

100

015

00

500

0

0 103 104 105102-102

CD3-PECy7

CD

4-B

V60

510

510

20

-10

2

103 105-102

104

103

1020 103 104

CD3-PECy7

CD

8-A

F70

010

510

410

30

-10

3

1020 103 104 105-102

CD3-PECy7

CD

56-P

E D

azzl

e10

510

410

3-1

02

1020 103 104 105-102

102


CD45: PerCP-Cy5.5 - Area

SSC

- A

rea

(x 1

0)

250

020

00

150

010

00

500

0

0 103 104 105 106-103


CD3: PE-Cy7 - Area

CD

4: B

V60

5 -

Are

a10

610

510

410

3-1

03

0

103 104 105 106-103

CD3: PE-Cy7 - Area

CD

8: A

lexa

Flu

or

700

- A

rea

106

105

104

0-1

04

0

103 104 105 106-103

CD3: PE-Cy7 - Area

CD

56: P

E-D

azzl

e594

- A

rea

106

105

104

0-1

03

0

103 104 105 106-103

103

CD45-PerCPCy5.5

SSC-

A(x

10

)30

00

250

020

00

100

015

00

500

0

0 103 104 105102-102

CD3-PECy7

CD

4-B

V60

510

510

20

-10

2

103 105-102

104

103

1020 103 104

CD3-PECy7

CD

8-A

F70

010

510

410

30

-10

3

1020 103 104 105-102

CD3-PECy7

CD

56-P

E D

azzl

e10

510

410

3-1

02

1020 103 104 105-102

102

1

3

5

2

4








Conventional Flow CytometerConventional Flow Cytometer



CD3: PE-Cy7 - Area

CD

19: A

PC

- A

rea

106

105

104

-10

40

103 104 105 106-103


CD3: PE-Cy7 - Area

HLA

DR

: AP

C-C

y7 -

Are

a10

610

510

410

3-1

03

103 104 105 106-103

CD3: PE-Cy7 - Area

TCR

-gd

: PE

- A

rea

106

105

104

103

-10

3

103 104 105 106-103

CD3: PE-Cy7 - Area

CD

27: B

V51

0 -

Are

a10

610

510

4-1

03

103 104 105 106-103

103

0

CD3-PECy7

CD

19-A

PC

105

104

103

102

-10

20

0 103 104 105102-102

CD3-PECy7

HLA

-DR

-AP

CCy7

105

102

0-1

02

103 105-102

104

103

1020 103 104

CD3-PECy7

TCR

GD

-PE

105

104

103

102

-10

3

1020 103 104 105-102

CD3-PECy7

CD

27-B

V51

010

510

410

3

1020 103 104 105-102

102

0


CD3: PE-Cy7 - Area

CD

19: A

PC

- A

rea

106

105

104

-10

40

103 104 105 106-103


CD3: PE-Cy7 - Area

HLA

DR

: AP

C-C

y7 -

Are

a10

610

510

410

3-1

03

103 104 105 106-103

CD3: PE-Cy7 - Area

TCR

-gd

: PE

- A

rea

106

105

104

103

-10

3

103 104 105 106-103

CD3: PE-Cy7 - Area

CD

27: B

V51

0 -

Are

a10

610

510

4-1

03

103 104 105 106-103

103

0

CD3-PECy7

CD

19-A

PC

105

104

103

102

-10

20

0 103 104 105102-102

CD3-PECy7

HLA

-DR

-AP

CCy7

105

102

0-1

02

103 105-102

104

103

1020 103 104

CD3-PECy7

TCR

GD

-PE

105

104

103

102

-10

3

1020 103 104 105-102

CD3-PECy7

CD

27-B

V51

010

510

410

3

1020 103 104 105-102

102

0


CD33: BV570 - Area

CD

27: B

V51

0 -

Are

a10

610

510

40

103 104 105 106-103


103

-10

3

CD33: BV570 - Area

CD

4: B

V60

5 -

Are

a10

610

510

410

3-1

03

103 104 105 106-103

CD33: BV570 - Area

TCR

-gd

: PE

- A

rea

106

105

104

103

-10

3

103 105 106-103

CD33: BV570 - Area

CD

56: P

E-D

azzl

e594

- A

rea

106

105

104

-10

3

104103 105 106-103

103

CD33-BV570

CD

27-B

V51

010

510

410

310

2

0 103 104 105102-102

0

CD33-BV570

CD

4-B

V60

510

510

20

-10

2

105-102

104

103

1020 103 104

CD33-BV570

TCR

GD

-PE

105

104

103

102

-10

2

0 103 104 105-103 102

CD33-BV570

CD

56-P

E D

azzl

e10

510

410

3

102 103 104 105-102

102

-10

2

0


CD33: BV570 - Area

CD

27: B

V51

0 -

Are

a10

610

510

40

103 104 105 106-103


103

-10

3

CD33: BV570 - Area

CD

4: B

V60

5 -

Are

a10

610

510

410

3-1

03

103 104 105 106-103

CD33: BV570 - Area

TCR

-gd

: PE

- A

rea

106

105

104

103

-10

3

103 105 106-103

CD33: BV570 - Area

CD

56: P

E-D

azzl

e594

- A

rea

106

105

104

-10

3

104103 105 106-103

103

CD33-BV570

CD

27-B

V51

010

510

410

310

2

0 103 104 105102-102

0

CD33-BV570

CD

4-B

V60

510

510

20

-10

2

105-102

104

103

1020 103 104

CD33-BV570

TCR

GD

-PE

105

104

103

102

-10

2

0 103 104 105-103 102

CD33-BV570

CD

56-P

E D

azzl

e10

510

410

3

102 103 104 105-102

102

-10

2

0


CD4: BV605 - Area CD4: BV605 - Area CD4: BV605 - Area CD4: BV605 - Area

CD

27: B

V51

0 -

Are

a10

610

510

40

103 104 105 106-103


103

-10

3

CD

19: A

PC

- A

rea

106

105

104

0-1

04

103 104 105 106-103

TCR

-gd

: PE

- A

rea

106

105

104

103

-10

3

103 105 106-103

CD

56: P

E-D

azzl

e59

4 -

Are

a10

610

510

4-1

03

104103 105 106-103

103

CD4-BV605 CD4-BV605 CD4-BV605 CD4-BV605

CD

27-B

V51

010

510

410

310

2

0 103 104 105102-102

0

CD

19-A

PC

105

102

-10

2

105-102

104

103

1020 103 104

TCR

GD

-PE

105

104

103

102

-10

2

0 103 104 105-103 102

CD

56-P

E D

azzl

e10

510

410

3

102 103 104 105-102

102

-10

2

0

15

F106

105

104

103

102

-101

-102

-102 -101 102 103

PE-Cy7_A

PE_

A

104 105 106

Cells

CD4_PacificBlue_A

CD

8_B

V4

21_A

-102 102

105

104

103

102

-1020

103

WLSM

A

GFP_A

-102

103

103

-102

-103

104

105

106

104

105

YFP

_A

Wavelength

800700600500

C

100

101

102

103

104

105

106

LD4

88

_A

Wavelength

800700600500

C

100

101

102

103

104

105

106

LD4

88

_A

Wavelength

800700600500

C

100

101

102

103

104

105

106

LD4

88

_A

Wavelength

800700600500

C

100

101

102

103

104

105

106

LD4

88

_A

WLSM

All events

Wavelength

LD48

8_H

800700600500

106

105

104

103

102

101

100

Common problems in flow cytometry occur when running fluorochromes with emission peaks that are too close to one another, multi-laser excitations, fluorescent proteins, and unstable tandems. The SP6800 is capable of analyzing all of these by looking at all photons from 420nm to 800nm and unmixing each unique spectral fingerprint.

Application Data: Dyes with issues

Example 1: Pacific Blue (457nm emission) and Brilliant Violet 421 (422nm emission)

Example 2: PE Cy5 (excited with 488nm and 638nm lasers) and APC (excited with 638nm laser)

Example 3: Fluorescent GFP and YFP need no special bandpass filter set with the SP6800

Example 4: Spectral Analysis unmixes the spectral fingerprint the spectral fingerprint of PE Cy7 tandem to produce excellent resolutions.

APC excitation

APC emission

PE Cy5 excitation

PE Cy5 emission

Normal PE Cy7

PE Cy7 falling apart

Unmixing

Wavelength (nm)

400 800700600500

Wavelength (nm)

400 800700600500

CD4_APC_A

-102 -101 102 103 104

106E

105

104

103

102

-101

-102

105

CD

8_P

E-C

y5_A

Sample Data

* Except for LE-SP6800ZC (This HPCV value is calculated for red laser. )

Configuration 2 laser model, 488/638 2 laser model, 405/488 3 laser model,405/488/638

Model Number LE-SP6800ZB LE-SP6800ZC LE-SP6800ZE

Optics

Laser specification

LasersSemiconductor 2 laser model488nm, 638nm

Semiconductor 2 laser model 405/488

Semiconductor 3 laser model, 405/488/638

Maximum power (at flow cell)488nm 40mW638nm 60mW

405nm 60mW488nm 40mW

405nm 60mW, 488nm 40mW 638nm 60mW

Irradiation form 2-spot dual-axis irradiation

Detection optics

Scatter signals Forward scatter (FSC), Side scatter (SSC)

Spectroscopic method Prism array spectroscopy

Fluorescent channels32 channel PMT(wavelength: 500-800nm)

32 channel PMT (wavelength: 500-800nm) PMT x 2 (420-440nm, 450-470nm)

System

Signal resolution Height 20 bit, Area 32 bit Sampling frequency: 50MHz

Measurement parameters64 channels, FSC, SSC, cell position XY

66 channels, FSC, SSC, cell position XY

Pulse shape parameters Height, Area, Width

Optical alignment Automated alignment with CorefinderTM technology

Event Rate 10,000eps (standard)/20,000eps (maximum)

Fluidics

Sample loader type Single auto-loader

Supporting sample tube 5ml polystyrene round tubes

Sheath Flow Speed Low (approx. 3m/s), Mid (approx. 5m/s), High (approx. 10m/s)

Detection channel dimensions 200μm x 200μm

Performance

Fluorescence sensitivity FITC: 120 MESF / PE: 70 MESF

Linearity FITC R2 ≧ 0.995 / PE R2 ≧ 0.995

Detection size range 0.5~40µm (beads)

Fluorescence detection resolution

488nm Laser/CH16, 638nm Laser/CH27* : ≧2.5% (HPCV)

Software

Unmixing algorithms Spectrum method (LSM, WLSM, PSA and Constraint option), Reverse matrix method (Conventional)

Autofluorescence Autofluorescence spectral detection

Virtual Filter Possible to change wavelength region for each fluorochromes in analysis

Export Data Format Flow Cytometry Standard (FCS) 3.0, 3.1

Main Unit

DimensionsMain unit: Width 60.0cm x Depth 63.5cm x Height 71.3cmFluidics cart: Width 78.6cm x Depth 52.1cm x Height 58.0cm

Weight Main unit: approx. 99kg (dried weight)/Fluidics cart: approx. 32kg (dried weight)

Air pressure supply 350kPa~450kPa (51psi~65psi)

AC Power supply AC100V 50/60Hz, AC120V 60Hz

Power consumption 220W (max)

Operating temperature 16-29 degrees Celsius, Room temperature variation: within 5 degrees Celsius

Operating humidity 20% to 80% (condensation free)

Recommended PC SP6800 workstation

Specifications

©2017 Sony Biotechnology Inc. All rights reserved. Sony, the Sony logo, and Flowpoint, are trademarks of Sony Corporation. eVolve is a trademark of and PE eFluor 610, eFluor 660, APC eFluor 780, and PerCP eFluor 710 are registered trademarks of eBioscience Corporation. Pacific Blue and Pacific Orange, are trademarks of and Alexa Fluor and Texas Red are registered trademarks of and licensed under patents assigned to Life Technologies Corporation. Qdot is a registered trademark of Invitrogen Corporation. Krome Orange, is a trademark of Beckman Coulter. BD Horizon, BD Horizon Brilliant and BB515 are trademarks of Becton Dickinson and Company. Brilliant Violet, Brilliant Violet 421, BV421, Brilliant Violet 570, BV570, Brilliant Violet 605, BV605, Brilliant Violet 650, and BV650, Brilliant Violet 510, BV510, Brilliant Violet 711, BV711, Brilliant Violet 785 and BV785 are trademarks of Sirigen Group Ltd. Vio is a registered trademark of Miltenyi Biotec GmbH. PE Dazzle 594 is a trademark of Biolegend. Cy and CyDye are registered trademarks of GE Healthcare. De Novo and FCS Express are trademarks of De Novo Software. All other trademarks are property of their respective owners. For non-clinical research use only. Not for use in diagnostic or therapeutic procedures, or for any other clinical purpose. The SP6800 Spectral Analyzer is a Class 1 laser product.

1.01.032916.4

1-7-1, Konan, Minato-Ku, Tokyo, 108-0075 JapanTel: +81 120-677-010 Fax: +81 [email protected]://www.sony.co.jp/LS

North America/International Japan Europe

1730 North First StreetSan Jose, CA 95112 U.S.A.Voice: +1 800-275-5963FAX: +1 [email protected]://www.sonybiotechnology.com

The Heights, Brooklands. Weybridge, Surrey, KT13 0XW, UK [email protected]

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