1. Microfluidic chip based flow cytometer
On-chip Sort
The world’s firstCell Sorter using disposable microfluidic chips for damage-free and sterile sorting.

2. FL4FL4FL4
LaserLaserLaser
chipchipchip
Flow pathFlow pathFlow path
Reflectingsurfaceby
totalinternalreflection
Target
Sort
PULL PUSH
Detection
80 µm
Non-target
Sample
SheathSheath
Waste
On-chip Innovation
Disposable chips: sterilized and cross−contamination freeMicrofluidic technology
Damage free sorting
Wide selection of sheath liquids and sample solutions
Flow system technology
Optical technology
Easy and maintenance-free operation
Fig 5. Flow and optics of microfluidic chip
Fig. 6. Principle of sorting
Fig . 3. The microfluidic chip enables a closed system
Fig. 2. On-chip Sort installed in biosafety cabinet
Microfluidic chip Micro flow paths
All fluids (sample, sheath and waste) stay within the chip during sorting and analysis, which prevents
contamination of the instrument. The Disposable micofluidic chips are just 5.5 x 4.0 cm with 80 x 80 µm
microfluidic channels. The chips are acrylic (PMMA), and are available as sterilized or unsterellized.
In addition, On-chip Sort is compact enough to fit inside a bio-safety cabinet, providing the best solution to deal
with biohazardous samples and strict safety regulations (Fig. 2).
Sheath and sample flow in micro channels are controlled
by air pressure. Fast liquid pulses of air pressure are used to
deflect target cells into the collection reservoir (Fig. 6).
Non-target cells continue in a straight line to the waste
reservoir, where they are fully recoverable as well.
Since sorting is dependent on fluid viscosity instead of ionic
strength, many different fluids can be used for the sample
and sheath fluids, including culture media, serum and
even oil.
Conventional cell sorters damage cells through high speed
collisions (10 m/s) in the collection vessel, high pressure
flow control (>45 PSI) which creates high shear stress in the
cells, and strong electric fields.
In contrast, On-chip Sorter causes no damage to the cells
because the flow speed is less than 1 m/s and the flow
control pressure is less than 0.3 PSI. Since electric fields are
not used, sample and sheath fluids are unconstrained by
ionic strength: users can choose buffers and media that
best suit their experimental needs.
Cells are detected in the laser irradiation area (detection
area in Fig. 6). On-chip software determines whether a cell
is a target cell while the cell approaches the sorting area.
When a target cell is passing through the sorting area,
push/pull pulse flow is generated for a few milliseconds to
send target cells into the isolation reservoir.
The electromagnetic valves and air pumps generate the
pulse flow and the technology is the solution for
damage-free, closed sorting.
The chip has two sheath flows which sandwich the sample
flow and narrow it to less than 10 µm in the middle of the
channel. This causes cells to pass through the flow cell one
by one in a line. Figure 4 is a cross section view of the chip,
with the flow path located in the middle.
Lasers irradiate from above the chip. Forward scatter (FSC)
and fluorescence are detected through an objective lens
below the chip (Fig. 5)
On the other hand, side scatter (SSC) and FL4 are
reflected by the edge slopes, and detected through
optical guides (Fig. 4).
Freedom from time-consuming maintenance and cleaning: The microfluidic chips are placed in a chip holder
and then loaded into the machine. The pumps and valves in On-chip provide air pressure but never contact the
sample or sheath fluid, so they don’t require cleaning. After discarding the chip at the end of analysis or sorting,
operation is terminated by simply turning off the power (Fig. 3).
Fig. 4. Detection of scattered light
and fluorescent light
Fig. 1. Disposable microfluidic chip
Chip for both analysis and sorting
Chip Loading the sample Set in the holder Installation Run

3. Analysis Sorting
2. Analysis using biohazardous materials
2. Circulating tumor cells (CTC) sorting by On-chip Sort for clinical trial.
Apoptosis assay Fungicidal capacity of Meropenem against
Analysis in a culture medium
Analysis in a culture medium
Detection of side population (SP) Live/Dead analysis of
Multicolor assay
Isolation of CTCs
CK and/or Vimentin positives CD45 negative
WBC TC
Bright field
Hoechst 33342(Nuclei)
Cytokeratin
Vimentin
CD45
Gate Sort
Assessment of an anti-HIV-1 drug candidate
Collaboration with Dr. OKADA Seiji, Center for AIDS Research, Kumamoto Univ.: 2012 Collaboration with Dr. WATANABE Masaru, Shizuoka Cancer Center : 2013
*
Collaboration with Drs. Koh & Watanabe, Shizuoka Cancer Center, and Drs. Koizumi & Uehara, National Cancer Center Research Institute
Collaboration with Dr. SUZUKI Ikuro, Tokyo University of Technology: 2011
OCBT Dr. ISHIGE Masayuki: 2013
OCBT Dr. YAMASHITA Namiko:2011OCBT Dr. YAMASHITA Namiko:2011
OCBT Dr. TAKEDA Kazuo:2009
KATOIII cells, UV exposure
APOPCYTO Annexin V-Azami-Green Apoptosis Detection Kit
(Comparison of On-chip Sort and CellSearch™ using EpCAM expression or non-expression cell lines)
AnnexinV
Annexin V-Azami-Green
PI
Negative
Q4
Q4
Q1
Q3
Q2
Negative
Positive
Q3
Negative
Positive
Live cells Apoptotic cells Dead cells
Q2
Positive
Cell line
KATO-III
EpCAM expression
Low
High
Detection rate
82.4±8.5%
93.8±5.5%
Detection rate
On-chip Flow CellSearch®
28.8%
74.7%
PC-14
A549
Null 96.0±8.5% 0.0%
Sorter A: flow late = 5, Nozzle = 70 µm,
cooling system = ON
On-chip Sort: pressure = 20 kPa,
room temperature
Cells from hippocampus, rat E18
Sorted by Sorter A or On-chip Sort
Pl staining Culture
Neuronal cells from rat hippocampus are delicate and sensitive to mechanical stresses. Below is a comparative experiment of
On-chip Sort to conventional jet-in-air sorter A.
An aliquot of the sorted cells were stained by PI to check viability and the remaining cells were cultured. PI staining, the
percentage of dead cells did not increase much in Sorter A nor in On-chip Sort compared to the cells without sorting.
Damage for jet-in-air sorted cells appeared as early as 3 days after cultivation (data not shown).
By the 7th day, neuronal cells not subjected to sorting or sorted by On-Chip Sort showed healthy, “neuron like” morphology and
extended axons. In contrast, the majority of the cells sorted by sorter A died by day 3, and almost all of them were dead by day 7.
We confirmed the finding is reproducible. Clearly, this damage-free sorting is strong advantage, and On-chip Sort is a pioneer in
this field.
Cancer cells can be profiled as cytokeratin (CK), EpCAM, and nuclear dye (7-AAD) positive and CD45 negative. On the
other hand, white blood cells are detected as CK and EpCAM negative, 7-AAD and CD45 positive. CTC were sorted by
On-chip Sort for subsequent gene analysis which can be used for personalized medicine*
.
Furthermore, sorting trials using clinical samples have been launched which aim to detect/sort CTCs by organ/cancer
specific antigens. Non-FDA approved.
Without sorting Sorter A On-chip Sort
Without sorting
Day 7
FS
PI
Day 7
FL
Sorter A On-chip Sort
1. General flow cytometric analysis by On-chip Flow/Sort 1. Cell sorting by On-chip Sort : Hippocampus cells from rat fetal brain
0 hr 3 hrs
Propidium lodide(PI)
pattern 1
PE/Cy5-CD4
(FL-5B)
FITC-CD3
(FL-2B)
PerCP/Cy5.5-CD14
(FL-5B)
FITC-CD3
(FL-2B)
FITC-CD3
(FL-2B)
SSC
PE-CD45
(FL-3B)
X(nM)
Uninfected Infected
6 dpi
HIV-DsRed
1 1000 100
Candidate compounds
11 dpi
APC/Cy7-CD8
(FL-6R)
PE/Cy7-CD19
(FL-6B)
Cytokeratin-FITC Adjacent channel
P2=CD3*CD4* T cells
P1=Lymphocyte P2=Monocytes
P3=Granulocytes
Isolated CTCs are observed under microscope.
Isolated cells were then subjected for sequencing to detect mutation.
P1=Lymphocytes
P3=CD3*CD8* T cells
PE-CD45
(FL-3B)
P4=T cells P5=B cells
pattern 2
APOPCYTO Annexin V-Azami-Green Apoptosis Detection Kit, MBL, Code No.4690
OCBT KAWASE Yoshie: 2013
AnnexinV-AzamiGreen
Pseudomonas
E. coli

4. Supporting a variety of fluorescent colors/high sensitivity Specifications
Violet
Violet
Blue
Green
FL1-V
FL1
FL1
FL2-V
FL2-B
FL2
FL3-V
FL3-B
FL3
FL4-V
FL4-B
FL3-B FL4-B
FL4
FL5-V
Sample: rainbow calibration particles (8 peaks)
FITC Signal intensity (510-550 nm)FL5-B
FL5-R
FL5
FL6-V
FL6-B
FL6-R
Red FL5-R FL6-R
FL6
400 450 500 550 600 650 700 750 800
FL2 FL3FL4 FL5 FL6
Blue Green Red
Product name
On-chip Sort HS
Equipment (On-chip Sort)
On-chip Sort HSG
On-chip Sort MS5
On-chip Sort MS5G
On-chip Sort MS6
On-chip Sort LS5
On-chip Sort LS3
Fluorescence sensitivity: FITC<200 MESF
Lasers
Blue Red Violet
Blue Green Violet
Blue Red
Blue Green
Blue Violet
Blue
Blue
The number of detectors
6
6
5
5
6
5
3
Catalogue No.
362S3001
362S3001G
252S3001
252S3001G
262S3001
152S3001
132S3001
Frequency
One to three lasers can be installed in On-chip Sort, Blue (488 nm) is required for scattering, Red (637
nm), Violet (405 nm) and Green (561 nm) lasers are available as additional options. The system
achieves as high as <200 MESF FITC. Up to six detectors cover a wide variety of fluorescent colors,
customized laser(s) are also available such as 785 nm.
Detection of individual signals from multiple lasers is available in a single
detector, and it enables smaller and inexpensive equipment.
600 MESF
Blank
Optics
Fluidics
Analysis and Sorting
Safety
Size and Weight
PC and Software
Supply
Data resolution
Pulse analysis
Detection wavelength
Fluorescent sensitivity
Size sensitivity
Measurement parameter
Laser
Flow-cell chip
Material of chip
Flow channel size
Flow speed
Sheath buffer
Volume of sample
Volume of sheath
Sorting method
Purity
Yield
Cell damage
Cross-contamination free
Sterilized
Pressure
Detection speed
Sorting speed
Start-up
Shutdown
Generation of aerosol
Size (W × H × W, mm)
Weight (kg)
PC and Software
OS
Data format
Power requirements
Power Consumption
4 decades, 18 bit ADC
Height, Area, Width
FL1 (445/20 nm), FL2 (543/22 nm), FL3 (591.5/40 nm)
FL4 (607/24 nm), FL5 (676/37 nm), FL6 (>710 nm)
< 200 MESF FITC
FSC < 0.5 m, SSC < 1.0 m
Forward scatter (FSC), Side scatter (SSC), 6 PMTs (up to 10 parameters)
Up to 3 Lasers from 405 nm, 488 nm, 561 nm, 637 nm, 785 nm
Disposable microfluidic chip
Acrylic (PMMA)
80 m × 80 m
500 mm/sec
Any media can be used
20-300 µL or up to 1 mL*
* You can load up to 1 mL on the chip, but sheath fluid runs out before you use up the sample. You can, however, refill the sheath fluid after taking out the chip.
4-9 mL
“Flow Shift” to generate pulse flow
> 95% (depends on cell concentration)
> 80%
No
Yes, because of the disposable chip
Yes
0.3 PSI
4,000 events/sec
100 targets/sec
5 minutes
10 sec (no cleanup of tubing necessary)
No
620 × 330 × 390
45 kg
Laptop PC
Windows 7 or 8, 64 bit
Own format and FCS 3.0
AC100−240 V, 50/60 Hz
< 240 VA

5. On-Chip USA
Phone: (877) 666-5426
Email: usa@onchip.co.jp
Web: on-chipbio.com
7098 Miratech Drive, Suite 100
San Diego, CA 92121
On-chip Biotechnologies Co., Ltd.
204 Venture Port,
2–24–16 Naka-cho, Koganei-city, Tokyo 184–0012, Japan