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  CellenONE

Automate cloning steps with CellenOne

OVERVIEW:


cellenONE® X1, an automated single cell dispensing system based on patented piezo-acoustic technology, allows precise cell deposition on awide range of microplates (96, 384, 1536) and microwell substrates. Most dispensing and microfluidic technologies follow Poisson distribution, which leads to multiple cells per position and low efficiency. cellenONE uses software-integrated visual feedback to ensure only single cells are deposited in every position.

With cellenONE® X1, cell sample is divided into droplets of identical volume。

CellenONE technical characteristics:


√Based on gentle and highly precise

piezo acoustic dispensing technology

√Automated optical monitoring of

cells inside dispenser’s nozzle。

√Machine learning steps to map

dispenser according to each sample。

√The single cells were highly active after distribution

√Cell position defines whether

single-cell conditions are fulfilled

and whether cells will be dispensed

in next drop formed                            

Technology principle:


We have developed a very simple process.  Using our glass capillary based PDC technology, aspirate a volume of cell suspension as low as a few micrometers, with practically no dead volume.  And then, using a patented dispensing/imaging technology, monitor each drop being dispensed.  Our PDC dispensing technology typically dispenses droplet volumes of 100-300 picoliter.  As each drop is about to be ejected, we monitor it for the presence of a single cell.  If one cell is present, we deposit it into your choice of target vessel.  It could be a microcell, a well of a micrometer plate, a slide, or anything else you fancy. If there are no cells in the droplet, or more than one cell, we recycle the droplet to guarantee no loos of your precious rare cells.

Application:


cell sorting

single-celled acquisition

single cell clone amplification

tissue specificity study

NGS Omics analysis

Application case

HuBBBTM:


HuBBBTM technology is a very powerful B cell immortalization system. However, data were limited due to successful cloning of only 10% of immortalized B cells. The combination of HuBBBTM and CellenONE® dramatically increases cloning efficiency up to 90%, thus providing significant time saving and ensuring identification of the very best clones

Cloning field:


Aim: Demonstrate ability to clone cells used in mAb development (e.g. CHO, Hybridoma)

Difficulty: Ensure monoclonality and viability of individually isolated and cultured cells

CellenONE approach: Monoclonality was verified by printing single cells onto microscope slides,Clonal recovery / viability was verified by dispensing single-clones into 96 well plate and counting n colonies/well after 7 days of culture。

FACS vs CellenONEX1:


1。Aim: Compare clonal recovery obtained with BD FACS Aria 3 and CellenONEX1

2.Cells: Transformed CHO cells with GFP negative reporter gene and gene of interest in fusion of antibiotic resistance gene. At time of experiment, 70% of CHOs were GFP negative.

3.Details: Cloning was undertaken in96wp format, wells were prefilled with 200μL culture media supplemented with selective antibiotic. After 5 days of culture, number of wells with one single colony with at least 5 cells were counted as positive. (GFP+ clones were eliminated due to selective antibiotic presence)

4。 Expected results:

i. 100% outgrowth after FACS sorting (excluding GFP +ve) 

ii. 70% outgrowth after CellenONE sorting (printing all cells, i.e. no sorting)

5。Actural results:

CellenONE technical information:


1.Dispensing technology: Piezo acoustic drop-on-demand

2.Dispense volume: 50-800 pL per drop

3.Linear for X/Y & Spindle for Z

4。Resolution: 1μm

5.Accuracy (Absolute Position):< 10μm

6。Precision (Repeat Position):< 3μm

7。Max。 speed:100厘米/秒

8.Spottable area (mm):x = 180;y = 120(2 microtiter plates)

9.Dimensions LxWxH (mm): 740 x 750 x 1580,weight 205kg

10。Voltage:110 V;220 V

 

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