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  CellMonitor

Cell Incubator With Automated Imaging & Monitoring

CellMonitor - System Overview

CellMonitor – Automated* Microscope in Incubator

System Overview

Automation of all functions

Fast screening

High resolution fluorescence and brightfield microscopy

Image recognition software

Integrated in cell culture incubator

Enviromental conditions control

Multiphoton microscopy and laser based cell manipulation** (on demand)

*developed by Fraunhofer IPM & FIT, ** developed by ROWIAK GmbH

Robot

Microscope-table

Microscope

Incubation rack

Transfer-position

Lock

Features of CellMonitor

High resolution microscopic imaging in brightfield and fluorescence mode

Image analysis of fluorescence signals

Monitoring of confluency and cell counting

Continuous documentation of cell growth parameters

Selection of transfected cell cultures and single cell colonies

Condition dependent system control

Constant quality control

CellMonitor – Microscope Configuration

Fully automated inverse microscope

Bright field and fluorescence modes

Hardware and software autofocus

Automated phase contrast

LED illumination at 530 nm and 464nm

Standard filter set: GFP (525nm), other filters on request

Integration of multiphoton microscopy possible

Transmitted light: LED

Fluorescence: reflected light illumination with LED's at a variety of wavelengths possible (e.g. 464nm, 488nm, 530nm)

Up to threee different filter sets

Objectives (standard outfit): 125x/0.04, 5x/0.12 with phase contrast, 40x/0.6 with phase contrast

Resolution down to 0.8µm

Sample carrier for microtiter plates

Autofocus: Fast hardware autofocus by optical triangulation for scanning, software autofocus for high resolution

Imaging speed: Complete scan of a 6 well MTP at 1.25x magnification ca. 200 images in ca. 4 min

CellMonitor – Microscope Specifications

MTP Handling

Loading of a MTP into the system

CellMonitor – Software Features

Standard Software Features

Analysis of cell confluency

Assessment of number of cells or confluent colonies

Reporting on position, roundness, and diameter of cells or cell colonies in the field of view

User adjustable cell recognition parameters

Standard software has been programmed to automatically recognize cells with certain optical performance i.e. transfected HeLa cells or mouse embryonic stem cells

Implementation of automatic recognition of cell types with other optical performance possible

Imaging Capabilities – Bright field

Sample images: Hela-Kyoto wild type

Cell cultures originate from the Max-Planck-Institut für Zellkultur und Genetik MPI-CBG

Brightfield 5x, phase contrast Brightfield 40x, phase contrast

Sample images

Cell cultures orginiate from Max-Planck-Institut für Zellkultur und Genetik MPI-CBG

Brightfield 40x Brightfield 40x, phase contrast

Fluo 40x, Autofluorescence

Imaging Capabilities – Bright field & Fluorescence

Transgenic ES-cells Transgenic ES-cells Transgenic ES-cells

Sample images

Imaging Capabilities – Multiphoton Mode

DAPIDAPI

Multiphoton image of HeLa cells

Connexin46-GFP Connexin46-GFP

3D reconstruction of 25µm Z-stack of HeLa cells

Sample images

Mosaic image of mouse vertebra (1x1mm)

Imaging Capabilities – Multiphoton Mode

Autofluorescence/SHG

Image Recognition Software

Confluency measurement

Self learning algorithms

User defines sample patterns for

1. Foreground

2. Background

Automated confluency measurement in high-throughput

Bright field mode

Recognized patterns

Image recognition software

Cell counting

Sample definition

Automatic identification of the remaining cells

Calculation of total cell number

Recognized number of cells

Illumination filtersTRAINED

Foreground/Background analysis

Extraction of individual regions

Selection of patterns based upon characteristics like diameter and morphology

Image Recognition – Colony Counting

Illumination filtersTRAINED

Foreground/Background analysis

Extraction of individual regions

Selection of specific

colonies

Image recognition – Fluorescence I

CellMonitor – Modular System Design

CellMonitor – Modularity

CellMonitor – Key Components

Fully automated microscope

Microscope incubator

Image recognition software

Multiphoton microscopy for imaging and manipulation (on request)

Additional modules can be added

Colony Picker

Air recirculation Laminar-Flow

Microscope incubator

Liquid-handling unit

refrigerator

Pickermodule

Storage incubator

Workbench

2D-Planar table as handling system driven by linear motors incl. runner

From CellMonitor to CellCultivator

Additional modules can be added to the Cell Monitor

Complete System - CellCultivator

Storage Incubator Grappler for MTP supply to the

microscope table

Automated positing system

Conditions: 37°C, > 95% rel., 5% CO2; incubator sterilizable

Outer dimensions (WxDxH): 110x80x186 cm

Inner dimensions (WxDxH): 76,8 cm x 67,8 cm x 74,6 cm

Weight: approx. 550 kg

Power consumption: ca. 2 kW

Capacity of Storage Incubator: 500 plates

Gate

Plate Feeder

CellMonitor - USPs

Optimized for sensitive cell cultures

Continuous maintenance of optimal conditions

Integration of multiphoton microscopy

Optimized monitoring of tissue culture for tissue engineering

Laser cell manipulation possible

Modular setup of complete system

Individual customized solutions possible

CellMonitor – Key Applications

Cultivation of stem cells

Cultivation of primary cells

Long term monitoring

Transfection by optoporation

Tissue engineering

3D culture

3D live cell imaging and manipulation

Imaging of tissue up to a high depth

© BD Biosciences

Suitable plate format

Connexin46-GFP expressing HeLa cells

Why 3D culture?

Mimicking biological and physiological conditions more realistic than monolayer cell cultures

“In vivo like Morphology”

Needed for tissue engineering and stem cell cultivation and differentiation

Might help to fill the gap between monolayer culture and animal trial in pharma

more predictive lead generation

more relevant ADMET/Tox results

Examples of three dimensional tissue models

Francesco Pampaloni, Ernst H. K. Stelzer and Andrea Masotti: Three-Dimensional Tissue Models for Drug Discovery and Toxicology; Recent Patents on Biotechnology 2009, 3, 103-117

CellMonitor – Marketing

CellMonitor – Target Markets

Pharmaceutical and Biotech Industry

Tissue Engineering

Pharmacology

Toxicology

Cytology

Academia

Research institutes which work on generation of cell cultures (primary cells, embryonic stem cells, etc.) with high throughput

Key WordsMolecular Pathology, ADME Tox, Cell Culture, Target Identification, Regenerative Studies, Plant Science, Embryology/IVF, Cell Culture and Maintenance

CellMonitor – Summary

Automated functions : Microscope control, autofocus, objective changer, phase rings, condenser, light source control, etc.

Integration into automated cell culture processes: Automated transfer of MTPs from and to robot systems

Climatization: Temperature, humidity and CO2 controlled

Image analysis: Confluency, cell counting, fluorescence signal analysis

Low maintenance: High power LED light sources

Customized solutions possible: Integration of laser manipulation for laser cell transfection or organelle ablation, multiphoton microscopy