iline F Capturing the Most From Your Suspension Cell Culture Process. In Real-Time

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1 iline F Capturing the Most From Your Suspension Cell Culture Process. In Real-Time

2 iline F Continuous suspension cell culture monitoring in a bioreactor The iline F microscope will change the way you monitor suspension cells. It is more time and cost effective than any other system available. The iline F is specifically designed to offer an in-line, label-free and non-invasive solution, using automated and continuous differential digital holographic microscopy to determine cell morphology at a single cell level. The iline F is directly connected to the bioreactor via an innovative fluidics system, BioConnect, avoiding sample consumption and reducing the risk of contamination. Measurements Cell counting Cell viability Cell clusters Cell morphology, up to 16 parameters recorded (a.o. circularity, perimeter, diameter...) The iline F, in-line, non-invasive, label-free cell monitoring The iline F monitors suspension cells in stirred bioreactors continuously and in real time, limiting the number of operations and improving reproducibility. The automated system offers user-independent reproducible quantitative 3D live cell imaging. We designed the iline F to help you track every cell in a reliable and cost efficient way. Spend your time on what matters most. Figure 1: iline F system, microscope & BioConnect iline F features No sampling, no staining, no toxic waste Full traceability at single cell level Discrimination between dead and living cells Cluster s information without disruption Continuous and real-time data acquisition BioConnect, a unique pump design for continuous monitoring Figure 2: BioConnect port The iline F microscope is connected to the bioreactor through a disposable and autoclavable closed loop fluidics system, the BioConnect, which is specifically designed to preserve cell integrity. BioConnect features Reusable pump engine Cell-friendly membrane pump working like a heart No sample consumption Automated flushing

3 OsOne, a powerful analysis software Advanced analysis & ease of use The iline F is delivered with OVIZIO s OsOne software package. Particularly convenient in use, OsOne acquires holographic fingerprints of each cell, computes the results and allows the user to focus on these results while offering all the tools required for a thorough investigation of the underlying data. Browser & individual cell attributes Displays the list of cells present and gives the ability to inspect each individual cell of your sample. Click on a cell to see its image and its morphological characteristics. Cell density and viability are measured and reported in nearly real-time by using a label-free method. Each individual cell can be observed and analyzed. A simple click on the image or on the dot plot simultaneously shows all data linked to that particular cell including up to 16 morphologic parameters (diameter, perimeter, surface, circularity, intensity, aggregate size...). Fully customizable, the measurement and analysis parameters can be easily configured to meet the user s needs. Images, videos or the advanced cell parameters can be exported with 2 or 3 clicks in OsOne. Your data can then be processed in another software such as Microsoft Excel or exported as a PDF report of your monitoring session. OsOne even offers the possibility to export data in standard imaging or cytometry formats. Intuitive interface design & wizard assistance Easy and intuitive access to your suspension cell culture monitoring by following a series of well-defined steps. Figure 5: Cell browser Continuous Data Recording Real-time data acquisition and recording. Shows graphs displaying the evolution of the viability and the total and viable cell density during the complete run. Figure 3: Start monitoring Live phase imaging & cell contours Highlights each cell contour. The cell viability can be visually evaluated via the mask displayed in the image overlay. Red for dead cells, green for viable cells, yellow for clusters. Figure 6: Continuous monitoring Advanced 3D Image Analysis 3D phase image observation of the cells provides information on volumes and morphological aspects. Figure 4: Phase image with overlays Figure 7: 3D view

4 How it works Cells are pumped out of the bioreactor and into the BioConnect via an innovative pumping system. Then the cells flow through the imaging device and flow back to the bioreactor after holograms of the cells have been captured. The holograms are continuously analysed to compute cell density and cell viability. Easy Set-up Figure 8: Startup guide Assemble & Autoclave 2 Start OsOne & Connect Pump 3 Insert in iline F, Monitoring Starts (automatically) Differential Digital Holographic Microscopy Grating inducing a spatial phase shift Label-free holographic fingerprint In digital holography, the scattered light beam from an illuminated object interferes with a reference beam on a CCD camera allowing for a 3D numerical reconstruction of the object. Differential digital holography is the next step of this basic technology, that enables an important size reduction of the instruments and an increased stability, so there is no frequent re-alignment of the beams required. Light source Object Beam 1 Beam 2 Mirror Mirror Interference Process CDD Figure 9: Differential Digital Holographic Microscopy principle

5 Specifications iline F Type Light source Analysis time iline-f Holographic Transmission Microscope Partially coherent monochromatic LED 630 nm (Red) < 30 s Cell size 2 µm to 100 µm Sensor Microscope objective Total magnification CCD camera 2456 (H) x 2058 (V) 8 bits Olympus LUCPLFLN20X Microscope Objective 20x 22.2 X Horizontal resolution 1 µm Vertical resolution (Optical Phase Variance) 2 nm Field of view 320 µm x 320 µm Acquisition rate Data volume Power consumption 15 fps 128 Mb per measurement point 100 W Integration Capabilities Automation via RestFUL web services OPC ready BioConnect Bioreactor compatibility Cell concentration range Throughput Cell travel time Tubing length Tubing diameter Tubing volume Retention volume Pump body volume Autoclave cycle Materials Flow cell dimensions Adaptable to most stirred tank bioreactors [0.1-20]*10 6 cells/ml above 20*10 6 cells/ml 4.8 ml/min Average 120 s 3.7 m (loop) Typical tubing size is O.D. 3 mm I.D. 1 mm. 3 ml 8 ml 5 ml Maximum 120 C 20 minutes USP v6.0, autoclavable, biocompatible silicon tubing. Silicone duckbill valves. Moulded Polycarbonate (Apec). Polycarbonate flow cell µm (W) x 200 µm (H) Dip tube Diameter (mm) Insertion length (mm) Tube material Stainless steel 316L Stainless steel 316L Max. Flow rate 12 ml/min Physical data Device 322 mm (L) x 212 mm (W) x 400 mm (H) 12 Kg

6 SALES AND SERVICE CONTACTS For more information on the product: Customer service: Manufactured by: Ovizio Ovizio Imaging Systems NV/SA 100, rue du Bourdon B-1180 Brussels / Belgium T E info@ovizio.com Find all Ovizio s microscopes Distributed by: Applikon Applikon Biotechnology B.V. Customer Service Department Heertjeslaan JG Delft / Netherlands T F E info@applikonbiotechnology.com Applikon US Headquarters 1180 Chess Drive Foster City, CA / U.S.A. T (650) F (650) E info@applikonbio.com