Intelligent Microplate Reader - hp-med. · PDF filemicroscopy with sensor-equipped test plates in an automated system HP Medizintechnik GmbH Fields of operation . ... (OCR) and Extracellular

Intelligent Microplate Reader

Product information

Manufacturer HP Medizintechnik GmbH

Located in Oberschleißheim (near Munich airport), Germany

Founded 2006

20 employees

More than 40 years of expertise in the manufacturing of laboratory equipment and medical devices

Developing, manufacturing and distributing of equipment for the sector of laboratory work, analysis and medicine technology

System integrator in the field “Modular and mobile reprocessing unit for medical devices"

Medical device act certification in 2010, with development and production structure for medical products

Steam sterilizers and autoclaves

We manufacture the

VARIOKLAV® autoclaves series steam sterilizers with quick loading and

simple operation, steam sterilizers with automatic

locking-systems and special equipment in the

medical sector

HP Medizintechnik GmbH Fields of operation

Modular and mobile reprocessing unit for

medical devices

Our modular systems for instrument reprocessing

secure medical care mobile contingents of the

medical service. Due to its mobility and flexibility

they can be used in many areas


Platforms for automated cell-based assays

Our biological analysis platform combines for the first time the light

microscopy with sensor-equipped test

plates in an automated system


Developments in cellular assays

Why

automate and

improve

cell-based

assay technology?


Cells react to their environment with changes in their physiology

Endpoint

analyses

This is traditionally detected by so called “endpoint analyses”


Sensor technology can overcome the problems of endpoint analysis

Endpoint

analyses

Sensor technology


Automation can control the environmental conditions

Endpoint

analyses

Sensor technology Control technology


Endpoint

analyses

Automated cell-based assays in real-time

Dynamic evaluation of multiple parameters simultaneously

Control Read out


Combined a powerful tool for cell-based assays is created

The whole cell can be understood as complex, non-linear system

Input signals can be controlled (e.g. drug application)

Why monitoring the whole cell?

The whole cell can be understood as complex, non-linear system

Input signals can be controlled (e.g. drug application)

Global cellular output signals represent the cellular behavior

Why monitoring the whole cell?

Physical Stress

Glucose

Oxygen

Viral infection Ligand-receptor activation

COOH OOCCH3

CH3

N

Active substances

H+

H+ H+

CO2

CO2

Oxygen consumption

Extracellular acidification

Cell morphology / count

The Intelligent Microplate Reader

The

Automated Monitoring System

for

Cell-based Assays

in the fields of

toxicology, diagnostics,

drug screening and research



Features of the measuring platform IMR

Analysis of the cellular physiology

Continuous measurement of pH-value, dissolved oxygen and impedance, plus microscopy

Calculation of Oxygen consumption rate (OCR) and Extracellular acidification rate (ECAR)

Parallel treatment and monitoring of 24 independent samples

Measurement of cells and tissues

Programmable imaging during the assays


Autonomous working over at least 3 days without the need to intervene by laboratory personnel

Bright-field and fluorescence microscopy

Automated media supply via a pipetting robot

7 different positions for media storage plates

Climate control and sterile technology

Possible injection of hypoxic gas


Automated data post-processing and display in real-time

All measured data is saved together with exact time-points

Data can be exported in common file formats like CSV and Jpeg for downstream data processing

Possibility of plausibility analysis by parallel recording of the same cell properties with different measuring methods (for example cell count over impedance and imaging)


The Intelligent Microplate Reader is certified as in-vitro diagnostic medical device according to the directive 98/79/EC

Overview of the IMR system components

Planar microscope light

XY-stage for microscope

Objective revolver

Sensor plate

Pipetting robot (24 channel)

Climate chamber

Automated inverse

microscope

The Sensor Plate: Heart of the IMR system

24 independent wells

Opto-chemical sensors for O2

and pH

Impedance sensing

Transparent carrier plate for Imaging

3-chamber fluidic system in well

Volume reduction cartridge (VRC)

pH O2

IMP

VRC

Different sensor plate types

4 different

sensor plate types

are available

The 3-chamber fluidic system in every well

Every well of the 24 well sensor plate is separated in 3 chambers

Metabolism sensors

Impedance sensors

Side chamber

Side chamber

Central chamber


Cells were seeded out in the central chamber

Metabolism sensors

Impedance sensors

Side chamber

Side chamber

Central chamber


Volume reduction cartridge

After attachment the central chamber volume is minimized


Volume reduction cartridge

The side chambers are now media ports to the cells


Exhausted medium is extracted by the robotic system


Fresh medium and drugs are added by the robotic system

High resulution imaging

Automated high resolution imaging gives you an accurate view of the cell morphology

50 µm

High resulution imaging

For generating deeper cellular information, a device

conversion to fluorescence microscopy is possible

50 µm

Automated raw-data conversion

Monitored raw-data

(dissolved oxygen, pH)

and time periods

are used to calculate

kinetic rates

Intelligent Microplate

Reader

RAW

RATE

How rates are calculated

Recording

of

raw-data

Raw-data

in

detail


Me

diu

m c

han

ge

Me

diu

m c

han

ge

Me

diu

m c

han

ge

Time-points

of

media-changes


Me

diu

m c

han

ge

Me

diu

m c

han

ge

ΔO2 /Δt

Regression-line

calculation

of rate data


Display

of

rate-kinetic




The IMR has a global software for user management, assay sequence control and real-time data display on the platform

The IMR Operation Software

Software interface for data display

Possibility to displayed complete data from all wells in real-time

Ox

yg

en

co

ns

um

pti

on

[%

/min

]

Time

Choice of wells, parameters and time periods to be displayed


Oxyg

en

co

ns

um

pti

on

[%

/min

]

Acid

ific

ati

on

[-p

H/m

in]

Time

Time

You can export your graphs in presentable quality


Oxyg

en

co

ns

um

pti

on

[%

/min

]

Acid

ific

ati

on

[-p

H/m

in]

Time

Time

Display of every taken culture image with exact time-point

10x objective


Time

All measured data is saved together with exact time-points and can be exported in common file formats like CSV and Jpeg for downstream

data processing

The IMR Operation Software

Preparing an experiment

How to prepare an experiment……


Working with the

platform requires a

S1 cell culture

laboratory for

sterile work


Choose your cell system…..

MCF7 L929 MDA and so on…

Full equipped Impedance/Imaging Metabolism/Imaging Imaging only

…and the sensor plate you need


Seed out your

cells into the

sensor plate


Prepare your

media and

substances to be tested


Close the volume

reduction cartridge

over the

attached cells


Insert the

sensor plate

into the

platform


Get your real-time data



Example for chemosensitivity

application

Chemosensitivity assay with MCF-7


Monitoring of MCF-7 human breast carcinoma cell line during treatment with different concentrations of the chemotherapeutic Chloroacetaldehyde (CAA)

Sensor plate type “full equipped”

Monitoring of oxygen consumption (OCR), extracellular acidification (ECAR), impedance and microscopic imaging (12h pre-treatment monitoring)

24h monitoring during treatment


Oxygen consumption rate (OCR)

Extracellular acidification rate (ECAR)

Impedance index

Imaging

Results overview:




Impedance index

Imaging

Comparable at every time point

Results overview:

10 µM CAA

Control (untreated)

Begin of treatment

25 µM CAA

n=7


Instant, dose-dependent and clear separated effect on mitochondrial system

Oxygen consumption rate

Wolf et al. Biosens. Bioelectron. 2013 Dec 15;50:111-7.

10 µM CAA

Control (untreated)

25 µM CAA

n=7


Time-depend and gradual effect on glycolysis system

Begin of treatment

Extracellular acidification rate


10 µM CAA

Control (untreated)

25 µM CAA

n=7


Cell degradation follows the effect on the glycolysis

system

Begin of treatment

Impedance index



Bright field imaging after 24h treatment


Clear morphologic degradation of cells at

highest dose

10x objective

Imaging

Example for toxicologic application

Toxicology assay of antiseptic care product


Monitoring of L929 mouse fibroblast cell line after short time treatment with different concentrations of an antiseptic care product

Sensor plate type “Metabolism/Imaging”

Monitoring of oxygen consumption (OCR), extracellular acidification (ECAR) and microscopic imaging (24h pre-treatment monitoring)

5 minute treatment (washing, application, washing; done by robot)

24h post-treatment-monitoring

Results overview:



Imaging


Results overview:



Imaging

Comparable at every time point



Automated 5 minute Treatment followed by washout

n=3

Instant, dose-dependent effect on mitochondrial

system

0.01% Antisept

Control (untreated)

0.10% Antisept

1.00% Antisept

Oxygen consumption rate


n=3

0.01% Antisept

Control (untreated)

0.10% Antisept

1.00% Antisept

Dose-dependent effect on

glycolysis system with cellular counter reaction at low dose

Extracellular acidification rate

Automated 5 minute Treatment followed by washout


Directly before 20 min after

Bright field imaging before and after treatment

Instant effect on cell

morphology

1% Substance

Control

10x objective

Imaging

If you don´t need extensive data, shorten your assays for faster results

Shorten your assay

Publications

The Intelligent Microplate Reader is utelised in different peer-reviewed scientific publications:

F. Demmel, M. Brischwein, P. Wolf, F. Huber, C. Pfister, B. Wolf: Nutrient depletion and metabolic profiles in breast carcinoma cell lines measured with a label-free platform. Physiol. Meas. 2015; 36: 1367–81

P. Wolf, M. Brischwein, R. Kleinhans, F. Demmel, T. Schwarzenberger, C. Pfister, B. Wolf: Automated platform for sensor-based monitoring and controlled assays of living cells and tissues. Biosens. Bioelectron. 2013; 50: 111-7

J. Wiest, A. Namias, C. Pfister, P. Wolf, F. Demmel, M. Brischwein: Data Processing in Cellular Microphysiometry. IEEE Trans. Biomed. Engineering 2016; 63: 2368-75

C. Pfister, C. Bozsak, P. Wolf, F. Demmel, M. Brischwein: Cell shape-dependent shear stress on adherent cells in a micro-physiologic system as revealed by FEM. Physiol. Meas. 2015; 36: 955-66

R. Kleinhans, M. Brischwein, P. Wang, B. Becker, F. Demmel, T. Schwarzenberger, M. Zottmann, P. Wolf, A. Niendorf, B. Wolf: Sensor-based cell and tissue screening for personalized cancer Chemotherapy. Med. Biol. Eng. Comput. 2012; 50: 117-26

T. Schwarzenberger, P. Wolf, M. Brischwein, R. Kleinhans, F. Demmel, A. Lechner, B. Becker, B. Wolf: Impedance sensor technology for cell-based assays in the framework of a high-content screening system. Physiological Measurement 2011; 32:977-93

P. Wolf, F. Hartl, M. Brischwein, B. Wolf: Determination of dynamic doxorubicin-EC50 value in an automated high-content workstation for cellular assays. Toxicology in Vitro 2011; 25(8):1889-94



Summary

.. overcome the problems of end-point assays

.. analyse the cellular physiology by continuous measurement of pH-value, dissolved oxygen and impedance in combination with microscopy imaging

.. calculation the Oxygen consumption rate (OCR) and Extracellular acidification rate (ECAR) in real-time

.. treat and monitor 24 independent samples in parallel

The Intelligent Microplate Reader can…

Summary

.. measure cells and tissues

.. supply media and substances automatically via a pipetting robot

.. process and display measured data in real-time

.. operate with 4 different sensor plate types

.. export all data for common data processing like StrataQuest analysis software

.. improve your investigation in toxicology, drug development and personalised medicine

The Intelligent Microplate Reader can…

Thank you for your attention

For further information visit us at www.hp-med.com

Documents

Intelligent Microplate Reader - hp-med. · PDF filemicroscopy with sensor-equipped test plates in an automated system HP Medizintechnik GmbH Fields of operation . ... (OCR) and Extracellular