CYTOGENETIC AUTOMATION Automated cell distributor on standard microscopic glass side for karyotyping and hybridization in situ Zendropper

Why the Zendropper? Automatisation of a very sensitive step High reproducibility of distribution from one patient to another and from one slide to another No inter-technician variations Homogeneisation and reliability of distribution Samples sparing Time saving and rapid execution

Why the Zendropper? Optimization of laboratory tasks System using a software allowing traceability of cells dispersion (bar- coded slides + session printing) Easier laboratory accreditation Better valorization of lab technician work Environmental factors under control ( T°, humidity, local aeration) with Water Layer methodology.

Zendropper methodologies Distribution on slide Caryotype Water Layer Methodology Distribution on slide Hybridization in situ Double cells spot on same slide Double cells spot on same slide Cells preparation

Sampling Cells culture Cells preparation Denaturation/Banding Microscope Reading Distribution on slide: ZenDropper 1. Karyotype procedure High automatic distribution capacity Homogeneous cells distribution 8 patients with 1 to 5 slides per patient (40 slides on 1 rack) Timing: (dispensing safety 100 %) +/- 25 ‘ for 40 slides  Significant time saving Protection of technicians not exposed to methanol and acetic acid emanation from Carroy solution.

Karyotype : Water Layer technology Distribution of a water layer on each slide Needle containing sample is placed just above the slide Creation of a central crater in the water layer due to Carroy solution evaporation Ejection of sample on the slide Aspiration of residual water on slide Slide ready for Banding step

Hybridation in situ Distribution of cell spot I 1 Distribution of cell spot II 2 3 Slide ready for next step Environment preparation : T°, Hygrometry

Advantages of water layer technology Improved mitosis thanks to contact water & acetic acid/methanol mix on slide surface  Chromosomes more spread  Banding (R or G) with better quality Creation of a micro-environnement  The distribution will always be done with same hygrometric conditions independently of climatic conditions Karyotype : Water Layer technology

Zendropper process 1Positioning and identification of samples 2Positioning standard microscopic slide 3Automatic validation by the Zendropper, of the sample presence and determination of sample volume (from 500µl to 4000µl]. 4Water dispensing on the slide for sample 1 5 Cells mixing following 3 programs of mixing defined automatically in point 3. 6Aspiration of sample volume needed to dispense on slides 7 Needle positioning above slide, waiting time to create central crater on water due to evaporation of Carroy solution and Sample ejection on slide

Zendropper process 8Aspiration of residual water on slide 9Next slide process 10Needle rinsing and next patient 11Informatic management of session and possibility to print session steps

Contamination : Protocol : Cells pellets of 240 patients have been distributed on routine microscopic slide alterning male/female patient Results : No chromosome Y observed on female samples and confirmed by Quinacrine test (100 mitosis per patient) Conclusion : No contamination between patients Comparative study Manual >< Zendropper CERBA Pasteur - Paris (Genetics departement - More than samples with Zendropper since March 2007) Zendropper process validation

Quality of mitosis : Less crossing of chromosomes with Zendropper Quantity of mitosis: More reproducible and homogenous dispersion of cells in mitosis with Zendropper Quality of « Banding » : High level of homogeneity from slide to slide with Zendropper Reading : Easier with Zendropper due to quality of dispersion Very efficient particularly for metaphasis research Lab technicians reactions: Easier and fast method than manual Less exposition to toxic vapours No more difficult work due to work position on bench or with a glove box. Zendropper process validation

Dimensions: 60x60x80 cm Weight: 62 kg Rack with slides : 8 patient X max 5 slides Tube holder for samples: 8 Extra washing needle solution: 2 positions 2 needles Needle 1 : - Mixing, aspiration and distribution of samples + distribution of water Needle 2 : Aspiration of residual water Technical Characteristics

Automate PC windows Xp Software for operations Software for setting up options of user and instrument calibrations (Ztoolbox) Printer 1 rack of 40 slides Option : Bar code reader Technical characteristics Zendropper system inclus

Zendropper Software Zendropper software Session preparation : number of samples, number of slide/patient (codebar available), print of session with information for tracability Ztoolbox Technical tool for maintenance and instrument set up in order to obtain high homogeneous cells dispersion conditions (Humidity and T°)

Intensive rinsing procedure at start and end of the day Regular replacement of needles and tubing due to intense corrosion of solution Zendropper maintenance Distribuito in ITALIA da Li StarFish S.r.l. Via Cavour, Cernusco S/N (MI) Tel Fax