SONY SP6800 (Cell Analyzer)
Spectral Analysis Technology
Lasers: Violet (405 nm), Blue (488 nm), Red (638 nm)(SONY, HP)

Spectral flow cytometer
1. Sum the fluorescence together
2. Use unmixing to mathematically separate the colors
Fluorescent dye(A+B+C+D)

1 cell data = 1 line

Fluorescentdye (A)

Fluorescentdye (B)

Fluorescentdye (C)

Fluorescentdye (D)

SONY SP6800 (Cell Analyzer)

It’s easy!

User

1. Turn on the power to start the SP6800(PC and SP6800 software)
2. Run “priming” and “Daily QC”
3. Create “Experiment”(You can select the colors you use)

＜First time＞
You need to register the spectral references.
4. Acquire measurement data and analyze data
5. Run “Daily cleaning”
6. Shut down
(SP software and PC)

SONY SP6800 (Cell Analyzer)
Advantage
No need for a single stain sample→ You can analyze small samples in multiple colors.
The wavelengths are close…
→ If the waveforms are different, staining is possible.

【Example】
・We collect the blood sample from mouse. (60 µl)
・We prepare these samples.
Negative control (30 µl) + Staining sample (30 µl)

CD19-BV421
CD45.2-PE
Gr-1-PerCPPI (dead cell)CD11b-FITC
CD45.1-APC
CD4/CD8-AF700

SONY SP6800 (Cell Analyzer)
Ogawa lab’s spectral reference list
BV421, EGFP, AF488, FITC

PE, PI, PerCP, tdTomato
APC, AF700, BV785

If you want to use these color references, feel free to contact us !!

Userload spectral reference → no need for single stain ^^SONY SP6800 (Cell Analyzer)
Advantage
No need for a single stain sample→ You can analyze small samples in multiple colors.
The wavelengths are close…
→ If the waveforms are different, staining is possible.

Disadvantage

Spectral flow cytometer sums the fluorescence together…
→ You cannot change the voltage of each color.

SONY SH800 (Cell Sorter)
Laser = 4
Violet (405 nm), Blue (488 nm), Yellow-Green (561 nm), Red (638 nm)

Detector = 6 (Choose the filter set you need)
450/50, 525/50, 585/30, 600/60, 617/30, 665/30, 720/60, 785/60(SONY, HP)【Example】

Filter Set 1

Filter Set 2

FL1

BV421

BV421

FL2

EGFP

FITC

FL3

tdTomato

PEFL47-AAD

PIFL5AF700

APCFL6APC-Cy7

AF700

SONY SH800 (Cell Sorter)
It’s fast! 1. Turn on the power to start the SH800(PC and SH800 software)It’s easy!

User
2. Follow the “Starting Wizard”(chip loading, laser & filter setting)
3. Run “Auto Calibration”(chip alignment, sort calibration)
4. Create “Experiment”(You can select the colors you use)
5. Acquire measurement data and analyze data
6. Follow the “Shut down Wizard”(software and hardware; including cleaning)

SONY SH800 (Cell Sorter)
Advantage & Disadvantage
Sorting Chip can be used for 24h→ Low risk of chip clogging.
→ High cost !! (¥3,000 / chip)
Plate sorting→ Ex) 17 cells were observed(20 cells were sorted into 24 well plate)
2-way sorting※ 4-way sorting → BD

6 detectors※ 7 colors or more → BD
Longer wavelengths：detection sensitivity is low !!
→ Ex) APC, AF700, etc…：highly expressed antigen

BD FACS in IMEG
BD FACS Aria III• 3 lasers; Violet (405), Blue (488) and Red (633)• 9 detectors(BD, HP)

BD FACS Aria SORP
• 4 (+1) lasers; Violet (405), Blue (488), Yellow-Green (561) and Red (633)(+ UV (355))• 11 detectors

BD FACS Aria IIu• 3 lasers; Violet (405), Blue (488) and Red (633)• 9 detectors

The advantages of BD FACS

Advantage• Spatially-separated spot
• 4-way sorting• The temperature controlling system

Spatially-separated beam spots

FITC

PE-Cy5.5
PE

Separated(BD FACS)

Cy5

PE-Cy7

APC-Cy7Single(SH800)https://www.bdj.co.jp/pdf/64-136-00.pdf

4-way sorting【Example】

The temperature controlling system

You can keep cells cells at any temperature !!

The disadvantage of BD FACS

Disadvantage• Manual setting is required✓Cleaning of Flow Cell
✓Nozzle insertion✓stream checking
etc....