Intelligent Microplate Reader - hp-med. · PDF filemicroscopy with sensor-equipped test plates...
Transcript of Intelligent Microplate Reader - hp-med. · PDF filemicroscopy with sensor-equipped test plates...
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
HP Medizintechnik GmbH Fields of operation
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
HP Medizintechnik GmbH Fields of operation
Endpoint
analyses
This is traditionally detected by so called “endpoint analyses”
Developments in cellular assays
Sensor technology can overcome the problems of endpoint analysis
Endpoint
analyses
Sensor technology
Developments in cellular assays
Automation can control the environmental conditions
Endpoint
analyses
Sensor technology Control technology
Developments in cellular assays
Endpoint
analyses
Automated cell-based assays in real-time
Dynamic evaluation of multiple parameters simultaneously
Control Read out
Developments in cellular assays
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
Features of the measuring platform IMR
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
Features of the measuring platform IMR
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)
Features of the measuring platform IMR
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
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
The 3-chamber fluidic system in every well
Cells were seeded out in the central chamber
Metabolism sensors
Impedance sensors
Side chamber
Side chamber
Central chamber
The 3-chamber fluidic system in every well
Volume reduction cartridge
After attachment the central chamber volume is minimized
The 3-chamber fluidic system in every well
Volume reduction cartridge
The side chambers are now media ports to the cells
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
Me
diu
m c
han
ge
Me
diu
m c
han
ge
Me
diu
m c
han
ge
Time-points
of
media-changes
How rates are calculated
Me
diu
m c
han
ge
Me
diu
m c
han
ge
ΔO2 /Δt
Regression-line
calculation
of rate data
How rates are calculated
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
Software interface for data display
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
Software interface for data display
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
Software interface for data display
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
Working with the
platform requires a
S1 cell culture
laboratory for
sterile work
Preparing an experiment
Choose your cell system…..
MCF7 L929 MDA and so on…
Full equipped Impedance/Imaging Metabolism/Imaging Imaging only
…and the sensor plate you need
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
Chemosensitivity assay with MCF-7
Oxygen consumption rate (OCR)
Extracellular acidification rate (ECAR)
Impedance index
Imaging
Results overview:
Chemosensitivity assay with MCF-7
Oxygen consumption rate (OCR)
Extracellular acidification rate (ECAR)
Impedance index
Imaging
Comparable at every time point
Results overview:
10 µM CAA
Control (untreated)
Begin of treatment
25 µM CAA
n=7
Chemosensitivity assay with MCF-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
Chemosensitivity assay with MCF-7
Time-depend and gradual effect on glycolysis system
Begin of treatment
Extracellular acidification rate
Wolf et al. Biosens. Bioelectron. 2013 Dec 15;50:111-7.
10 µM CAA
Control (untreated)
25 µM CAA
n=7
Chemosensitivity assay with MCF-7
Cell degradation follows the effect on the glycolysis
system
Begin of treatment
Impedance index
Wolf et al. Biosens. Bioelectron. 2013 Dec 15;50:111-7.
Chemosensitivity assay with MCF-7
Bright field imaging after 24h treatment
Wolf et al. Biosens. Bioelectron. 2013 Dec 15;50:111-7.
Clear morphologic degradation of cells at
highest dose
10x objective
Imaging
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:
Oxygen consumption rate (OCR)
Extracellular acidification rate (ECAR)
Imaging
Toxicology assay of antiseptic care product
Results overview:
Oxygen consumption rate (OCR)
Extracellular acidification rate (ECAR)
Imaging
Comparable at every time point
Toxicology assay of antiseptic care product
Toxicology assay of antiseptic care product
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
Toxicology assay of antiseptic care product
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
Toxicology assay of antiseptic care product
Directly before 20 min after
Bright field imaging before and after treatment
Instant effect on cell
morphology
1% Substance
Control
10x objective
Imaging
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…