Streamline your bacterial growth assays
Transcript of Streamline your bacterial growth assays
Headquarters GermanyBMG LABTECH GmbHAllmendgrün 877799 OrtenbergTel. +49 781 96968 [email protected]
AustraliaBMG LABTECH Pty. Ltd.2/24 Carbine WayMornington, Victoria, 3931Tel. +61 3 5973 [email protected]
FranceBMG LABTECH SARL 7, Rue Roland Martin94500 Champigny s/MarneTel. +33 1 48 86 20 [email protected]
JapanBMG LABTECH JAPAN Ltd.1-6-2, Shimo-choOmiya-ku330-0844 Saitama City Tel. +81 48 647 [email protected]
UK BMG LABTECH Ltd. 8 Bell Business ParkSmeaton CloseAylesburyBucksHP19 8JRTel. +44 1296 [email protected]
USABMG LABTECH Inc.13000 Weston ParkwaySuite 109Cary, NC 27513Tel. +1 877 264 [email protected]
www.bmglabtech.com
04/2
019
Made in Germany
Streamline your bacterial growth assays
SummaryThe bacterium Campylobacter jejuni causes food poisoning with symptoms of abdominal pain, diarrhea and fever. C. jejuni is cultured at 37-42°C, 5-8% O2 and 10% CO2 as this mimics conditions found in the intestine of its hosts.Here, a comparison of C. jejuni growth studies performed in 8 ml scale were compared with growth curves obtained in a 96 well microplate. The atmosphere was controlled with a VAIN workstation or the Atmospheric Control Unit of a micro-plate reader, respectively. Optical density at 600 nm to assess biomass was acquired in a cuvette or directly in the microplate.The comparison of growth of Campylobacter in tubes and in 96 well plates demonstrates that this fastidious organism can be transferred to a 96 well format. This is made easy by the design of the BMG LABTECH microplate reader providing temperature control, shaking and atmospheric control.
Growth of Campylobacter using a microplate reader equipped with ACUR.D. Haigh, J.M. Ketley Department of Genetics, University of Leicester
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
OD 6
00 n
m
0 2 4 6 8 10 12 14 16 18 20 22
Time (hours)
111688117681116
Growth of Campylobacter isolates over a 24 hour period in theFLUOstar® Omega.
Shak
ing
Exte
nsive
sha
king
Mul
ti-m
ode
dete
ctio
n
Incu
batio
n
Wel
l sca
ns
Atm
osph
eric
Con
trol
Uni
t
Wha
t els
e sh
ould
you
kno
w
SPEC
TROs
tar®
Nan
o
Our i
nnov
ative
abs
orba
nce-
only
mic
ropl
ate
read
er h
as th
e fl e
xibi
lity
to p
erfo
rm a
ssay
s qu
ickl
y an
d ea
sily
in m
icro
plat
es o
r via
the
built
in c
uvet
te p
ort.
This
spe
ctro
met
er-
base
d ab
sorb
ance
pla
te re
ader
cap
ture
s a
full
UV/
vis s
pect
rum
in le
ss th
an 1
sec
/wel
l. Its
spe
ed a
nd s
impl
e pu
sh b
utto
n op
erat
ion
mak
e it
the
lead
ing
mic
ropl
ate
read
er fo
r ab
sorb
ance
mea
sure
men
ts.
Omeg
a Se
ries
(Up
to 6
5°C)
The
Omeg
a se
ries
offe
r tru
e fl e
xibi
lity
to
prov
ide
you
with
the
perf
ect r
eade
r to
fulfi
l yo
ur re
quire
men
ts: f
rom
an
abso
rban
ce-
only
SPE
CTRO
star
or l
umin
esce
nce-
only
LU
MIs
tar t
o a
fully
-equ
ippe
d PO
LARs
tar
Omeg
a w
ith u
p to
sev
en d
etec
tion
mod
es.
Your
cho
sen
inst
rum
ent c
an a
lso
be
upgr
aded
at a
ny ti
me
if yo
u ne
ed m
ore
fea-
ture
s or
add
ition
al d
etec
tion
mod
es.
CLAR
IOst
ar®
Plu
s
(Mon
ochr
omat
or)
(Up
to 6
5°C)
The
CLAR
IOst
ar P
lus
is th
e id
eal i
nstr
u-m
ent f
or a
ssay
dev
elop
men
t. Ou
r mos
t fl e
xibl
e m
ulti-
mod
e pl
ate
read
er c
omes
eq
uipp
ed w
ith o
ur p
aten
ted
LVF
Mon
ochr
o-m
ator
s™, fi
lter
s, a
nd s
pect
rom
eter
. Fle
xibi
-lit
y co
mbi
ned
with
the
best
sen
sitiv
ity o
f its
cl
ass,
and
our
new
ly d
evel
oped
Enh
ance
d Dy
nam
ic R
ange
tech
nolo
gy, m
ake
it th
e id
eal r
eade
r for
ass
ay d
evel
opm
ent.
SummaryStreptococcus agalactiae (also known as group B streptococcus [GBS]) is associated with various diseases such as neonatal disease, sepsis, arthritis, pneumonia, meningitis, skin and softtissue infections. Host colonization and virulence are mainly studiedusing microscopy and fluorescent biomarkers. Strategies forlabelling GBS with fluorescent biomarkers have so far been limited to antibody-based immunostaining methods and non-specific protein/DNA stains.Green fluorescent protein (GFP) expression is a common labeling method for bacteria, enabling their identification in complex samples or monitoring of their subcellular locations in eukaryotic host cells. Recently, stable expression of a green fluorescent protein mutant (GFPmut3) with enhanced fluorescence intensityin GBS was reported. Validation and stability of GFPmut3 expression in GBS cultures was measured by the CLARIOstar multi-mode plate reader and its fluorescence intensity, fluore-scence polarization and absorbance measurement capabilities.
Expression of a stable GFP mutant in Group B Streptococcus. Growth, detection and monitoring.Matthew J. Sullivan and Glen C. Ulett, School of Medical Science, and Menzies Health Institute Queensland, Griffith University, Gold Coast, Queensland, Australia
1.00
0.75
0.50
0.25
0.000 3 6 9 12
Abso
rban
ce (O
D 600n
m)
Time (h)
A B3
2
1
0
-10 3 6 9 12
105
RFU
(GFP
FP
inte
nsity
)
Time (h)
Absorbance (A) and polarized fl uorescence intensity profi le (B) of GFP+ GBS (green) and GFP- GBS (black) grown in CDM media.
For more information please refer to BMG LABTECH application note 217.
For more information please refer to BMG LABTECH application note 329.
Minimise operational timeBMG LABTECH offers single and multi-mode microplate readers extensively used by researchers in microbiology. Whether studying microbiomes, biofilms or synthetic biology, plate readers can help expand your experimental possibilities. The classical bacterial experiment was to acquire a growth curve by drawing aliquots and measuring absorbance at 600 nmin a cuvette. Our instruments provide streamlined solutions enabling a modern approach to monitor growth of multiple samples (6–384 wells) in real-time with no manual intervention.
Increase your experimental outputOur plate readers provide a true “walk-away” solution that reduces user intervention and saves time. Instruments allow fast, automated kinetic detection spanning from seconds up to days. Besides absorbance, our multi-mode readers can detect fluorescent and luminescent signals in the same run. This provides the capability to normalise gene expression to cell density or measure cell growth in relation to O2 consumption. For higher throughput, all readers are robot compatible and can be easily automated with all of the leading platforms.
Replicate specific growth conditionsAccurate temperature control to 65°C, multiple shaking options with various modes and speeds and the capability to precisely regulate O2 and CO2 concentrations in the reader provide the best growth conditions even for the most fastidious microbes.
Customer service, reliability and savingsFree application support for the life of the instrument, and access to our knowledge and expertise ensure the maximum performance of your instrument and the best data from your experiments. With BMG LABTECH, “Made-in-Germany” depen-dability is by design. Our philosophy is that all instruments must be of the highest standard of reliability and functionality. This makes our readers the lowest cost of ownership and highest brand satisfaction on the market.
MultiplexingIf you wish to analyse multiple parameters, measurement ofOD600 can be combined with luminescent or fluorescent detection of up to 5 different dyes in the same run. This allows the design and performance of multiplex assays. For instance, the growth of multiple strains with different fluorescently tagged genes can be monitored in the same well, or the growth characteristics and effects from various bioactives can be followed in a single well. For even more flexibility the patented LVF-MonochromatorTM
allows you to select any wavelength and any bandwidth for your fluorescent or luminescent measurement, providing your lab with the most versatile reader available.
Temperature regulationMicroorganisms have different optimal temperatures for maximum growth rates. To ensure optimal growth conditions, BMG LABTECH readers offer accurate temperature regulation up to 65°C. The specific incubation zone guarantees a uniform temperature and minimizes evaporation. Temperature values are logged to the signal curve, making it simple to compare events at different temperatures.
ShakingMost microbes require aeration to ensure growth or to disperse cultures into an even suspension. Three shaking modes as well as adjustable speed up to 700 rpm or optionally to 1100 rpmprovide optimum settings for your strain. Additionally, the readers can be equipped with an extraordinary robust transport system for shaking 24/7 where required.
Orbital, spiral averaging, and matrix scan
Well scansMicrobial assays are most often non-homogenous. Hence, yourexperiments may benefit from a signal captured over the whole well-surface in either a spiral or orbital pattern, ensuring the most accurate measurement. More detailed matrix scan options (from 2x2 to 30x30) acquire multiple read points over the well surface and display the signal variation in a matrix. This is extremely advantageous when studying highly non-homogenous samples, such as flocculating bacteria or yeast, and for biofilm forming bacteria, to study biofilm size and shape in more detail.
SummaryBacteria communicate by producing, detecting and responding to low molecular mass molecules known as autoinducers (AI) in a process called quorum sensing. The process is well-described in Vibrio fischeri, an organism that turns bioluminescent with high cell density. The molecule responsible for bioluminescence development is N-3-oxo-hexanoyl-L-homoserine lactone (3-oxo-C6-HSL).A BMG LABTECH microplate reader detected biolumine-scence of V. fischeri to report on quorum sensing and relatedit to the cell density determined by OD600 measurements.V. fischeri strains mutated in genes required for lumi-nescence development were exposed to 3-oxo-C6-HSL tostudy cell-to-cell communication. Combining luminescence and absorbance measurements enabled to monitor microbial growth at the same time with bioluminescence. This means that a specific signal and an internal control can be acquired to allow a ratiometric read-out.
Monitoring bacterial cell-to-cell communication “quorum sensing” using a BMG LABTECH microplate readerK.E. Eboigbodin and G.K. Robinson Dept. of Biosciences, University of Kent, Canterbury, CT2 7NJ
3E+4
2.5E+4
2E+4
1.5E+4
1E+4
5E+3
0
RLU
/OD
(600
nm)
0 5 10 15 20 25
Time (h)
KV267-KV240-ESR1-
KV267+KV240+ESR1+
Normalized bioluminescence of V. fi scheri strains. Bacteria were either supplemented with exogenous HSL (+) or not (-). Insert shows growth curves
“After extensive testing of five different brands of microplate readers, only BMG LABTECH instruments were found to have the sensitivity, durability and reliability required for our applications.”
Associate Professor Christopher McDevitt, Group Leader The Peter Doherty Institute for Infection and Immunity,
The University of Melbourne, Australia
Key features to keep both microbes and researchers happy
Atmospheric Control UnitThe living environment of microorganisms is diverse; replicating it in a plate reader requires precise control of the atmosphere. The Atmospheric Control Unit (ACU) independently regulates O2 and CO2 inside the reader to establish an environment conducive for growth. This allows long-term measurements atstable O2 (0.1-20 %) and CO2 (0.1-20 %) atmospheres, perfect forthe most fastidious organisms. Gas ramping can be programmed to study responses to changing gas conditions in real-time.
ACU perfectly regulates both 02 and CO2
Linear, orbital and double-orbital shaking mode
Simplifi ed schematic of the LVF Monochromator technology.
SummaryIt is well known that microorganisms need a specific level of carbon dioxide (CO2) for growth and reproduction. The period during which this level is increased usually corresponds to the lag phase as the organism is unable to divide until the critical concentration of CO2 is reached. Neisseria meningitidis
is considered to require or benefit from a concentration of CO2
greater than atmospheric, hence it is a suitable organism to study CO2 effects.A strain of Neisseria meningitidis was used to assess the efficiency of a BMG LABTECH multi-mode plate reader coupled with an Atmospheric Control Unit (ACU) to deliver 5 % CO2. Growth was determined by detecting absorbance at 405 nm and compared to data obtained on a microplate reader without ACU.The BMG LABTECH plate reader with ACU is able to achieve and maintain a level of CO2 required for such a fastidious organism or for cell cultures.
Growth of Neisseria meningitidis in a BMG LABTECH microplate reader with Atmospheric Control Unit (ACU)Kerry L. Cutter University of the West of England, Bristol
100
90
80
70
60
50
40
30
20
10
0
% B
acte
rial G
row
th
0 5 10 15 20 25
Neat1 in 101 in 1001 in 10001 in 10,0001 in 100,0001 in 1,000,000
Time (hours)
Growth of serially diluted cultures of Neisseria meningitidis in BHI broth supplemented with 10 % FBS using a BMG LABTECH plate reader with ACU set to deliver 5 % CO2 at 37°C. The data presented were calculated from triplicate optical density readings (at 405nm) taken hourly over a 24 h period from duplicate experiments.
For more information please refer to BMG LABTECH application note 155.
For more information please refer to BMG LABTECH application note 199.
Minimise operational timeBMG LABTECH offers single and multi-mode microplate readers extensively used by researchers in microbiology. Whether studying microbiomes, biofilms or synthetic biology, plate readers can help expand your experimental possibilities. The classical bacterial experiment was to acquire a growth curve by drawing aliquots and measuring absorbance at 600 nmin a cuvette. Our instruments provide streamlined solutions enabling a modern approach to monitor growth of multiple samples (6–384 wells) in real-time with no manual intervention.
Increase your experimental outputOur plate readers provide a true “walk-away” solution that reduces user intervention and saves time. Instruments allow fast, automated kinetic detection spanning from seconds up to days. Besides absorbance, our multi-mode readers can detect fluorescent and luminescent signals in the same run. This provides the capability to normalise gene expression to cell density or measure cell growth in relation to O2 consumption. For higher throughput, all readers are robot compatible and can be easily automated with all of the leading platforms.
Replicate specific growth conditionsAccurate temperature control to 65°C, multiple shaking options with various modes and speeds and the capability to precisely regulate O2 and CO2 concentrations in the reader provide the best growth conditions even for the most fastidious microbes.
Customer service, reliability and savingsFree application support for the life of the instrument, and access to our knowledge and expertise ensure the maximum performance of your instrument and the best data from your experiments. With BMG LABTECH, “Made-in-Germany” depen-dability is by design. Our philosophy is that all instruments must be of the highest standard of reliability and functionality. This makes our readers the lowest cost of ownership and highest brand satisfaction on the market.
MultiplexingIf you wish to analyse multiple parameters, measurement ofOD600 can be combined with luminescent or fluorescent detection of up to 5 different dyes in the same run. This allows the design and performance of multiplex assays. For instance, the growth of multiple strains with different fluorescently tagged genes can be monitored in the same well, or the growth characteristics and effects from various bioactives can be followed in a single well. For even more flexibility the patented LVF-MonochromatorTM
allows you to select any wavelength and any bandwidth for your fluorescent or luminescent measurement, providing your lab with the most versatile reader available.
Temperature regulationMicroorganisms have different optimal temperatures for maximum growth rates. To ensure optimal growth conditions, BMG LABTECH readers offer accurate temperature regulation up to 65°C. The specific incubation zone guarantees a uniform temperature and minimizes evaporation. Temperature values are logged to the signal curve, making it simple to compare events at different temperatures.
ShakingMost microbes require aeration to ensure growth or to disperse cultures into an even suspension. Three shaking modes as well as adjustable speed up to 700 rpm or optionally to 1100 rpmprovide optimum settings for your strain. Additionally, the readers can be equipped with an extraordinary robust transport system for shaking 24/7 where required.
Orbital, spiral averaging, and matrix scan
Well scansMicrobial assays are most often non-homogenous. Hence, yourexperiments may benefit from a signal captured over the whole well-surface in either a spiral or orbital pattern, ensuring the most accurate measurement. More detailed matrix scan options (from 2x2 to 30x30) acquire multiple read points over the well surface and display the signal variation in a matrix. This is extremely advantageous when studying highly non-homogenous samples, such as flocculating bacteria or yeast, and for biofilm forming bacteria, to study biofilm size and shape in more detail.
SummaryBacteria communicate by producing, detecting and responding to low molecular mass molecules known as autoinducers (AI) in a process called quorum sensing. The process is well-described in Vibrio fischeri, an organism that turns bioluminescent with high cell density. The molecule responsible for bioluminescence development is N-3-oxo-hexanoyl-L-homoserine lactone (3-oxo-C6-HSL).A BMG LABTECH microplate reader detected biolumine-scence of V. fischeri to report on quorum sensing and relatedit to the cell density determined by OD600 measurements.V. fischeri strains mutated in genes required for lumi-nescence development were exposed to 3-oxo-C6-HSL tostudy cell-to-cell communication. Combining luminescence and absorbance measurements enabled to monitor microbial growth at the same time with bioluminescence. This means that a specific signal and an internal control can be acquired to allow a ratiometric read-out.
Monitoring bacterial cell-to-cell communication “quorum sensing” using a BMG LABTECH microplate readerK.E. Eboigbodin and G.K. Robinson Dept. of Biosciences, University of Kent, Canterbury, CT2 7NJ
3E+4
2.5E+4
2E+4
1.5E+4
1E+4
5E+3
0
RLU
/OD
(600
nm)
0 5 10 15 20 25
Time (h)
KV267-KV240-ESR1-
KV267+KV240+ESR1+
Normalized bioluminescence of V. fi scheri strains. Bacteria were either supplemented with exogenous HSL (+) or not (-). Insert shows growth curves
“After extensive testing of five different brands of microplate readers, only BMG LABTECH instruments were found to have the sensitivity, durability and reliability required for our applications.”
Associate Professor Christopher McDevitt, Group Leader The Peter Doherty Institute for Infection and Immunity,
The University of Melbourne, Australia
Key features to keep both microbes and researchers happy
Atmospheric Control UnitThe living environment of microorganisms is diverse; replicating it in a plate reader requires precise control of the atmosphere. The Atmospheric Control Unit (ACU) independently regulates O2 and CO2 inside the reader to establish an environment conducive for growth. This allows long-term measurements atstable O2 (0.1-20 %) and CO2 (0.1-20 %) atmospheres, perfect forthe most fastidious organisms. Gas ramping can be programmed to study responses to changing gas conditions in real-time.
ACU perfectly regulates both 02 and CO2
Linear, orbital and double-orbital shaking mode
Simplifi ed schematic of the LVF Monochromator technology.
SummaryIt is well known that microorganisms need a specific level of carbon dioxide (CO2) for growth and reproduction. The period during which this level is increased usually corresponds to the lag phase as the organism is unable to divide until the critical concentration of CO2 is reached. Neisseria meningitidis
is considered to require or benefit from a concentration of CO2
greater than atmospheric, hence it is a suitable organism to study CO2 effects.A strain of Neisseria meningitidis was used to assess the efficiency of a BMG LABTECH multi-mode plate reader coupled with an Atmospheric Control Unit (ACU) to deliver 5 % CO2. Growth was determined by detecting absorbance at 405 nm and compared to data obtained on a microplate reader without ACU.The BMG LABTECH plate reader with ACU is able to achieve and maintain a level of CO2 required for such a fastidious organism or for cell cultures.
Growth of Neisseria meningitidis in a BMG LABTECH microplate reader with Atmospheric Control Unit (ACU)Kerry L. Cutter University of the West of England, Bristol
100
90
80
70
60
50
40
30
20
10
0
% B
acte
rial G
row
th
0 5 10 15 20 25
Neat1 in 101 in 1001 in 10001 in 10,0001 in 100,0001 in 1,000,000
Time (hours)
Growth of serially diluted cultures of Neisseria meningitidis in BHI broth supplemented with 10 % FBS using a BMG LABTECH plate reader with ACU set to deliver 5 % CO2 at 37°C. The data presented were calculated from triplicate optical density readings (at 405nm) taken hourly over a 24 h period from duplicate experiments.
For more information please refer to BMG LABTECH application note 155.
For more information please refer to BMG LABTECH application note 199.
Minimise operational timeBMG LABTECH offers single and multi-mode microplate readers extensively used by researchers in microbiology. Whether studying microbiomes, biofilms or synthetic biology, plate readers can help expand your experimental possibilities. The classical bacterial experiment was to acquire a growth curve by drawing aliquots and measuring absorbance at 600 nmin a cuvette. Our instruments provide streamlined solutions enabling a modern approach to monitor growth of multiple samples (6–384 wells) in real-time with no manual intervention.
Increase your experimental outputOur plate readers provide a true “walk-away” solution that reduces user intervention and saves time. Instruments allow fast, automated kinetic detection spanning from seconds up to days. Besides absorbance, our multi-mode readers can detect fluorescent and luminescent signals in the same run. This provides the capability to normalise gene expression to cell density or measure cell growth in relation to O2 consumption. For higher throughput, all readers are robot compatible and can be easily automated with all of the leading platforms.
Replicate specific growth conditionsAccurate temperature control to 65°C, multiple shaking options with various modes and speeds and the capability to precisely regulate O2 and CO2 concentrations in the reader provide the best growth conditions even for the most fastidious microbes.
Customer service, reliability and savingsFree application support for the life of the instrument, and access to our knowledge and expertise ensure the maximum performance of your instrument and the best data from your experiments. With BMG LABTECH, “Made-in-Germany” depen-dability is by design. Our philosophy is that all instruments must be of the highest standard of reliability and functionality. This makes our readers the lowest cost of ownership and highest brand satisfaction on the market.
MultiplexingIf you wish to analyse multiple parameters, measurement ofOD600 can be combined with luminescent or fluorescent detection of up to 5 different dyes in the same run. This allows the design and performance of multiplex assays. For instance, the growth of multiple strains with different fluorescently tagged genes can be monitored in the same well, or the growth characteristics and effects from various bioactives can be followed in a single well. For even more flexibility the patented LVF-MonochromatorTM
allows you to select any wavelength and any bandwidth for your fluorescent or luminescent measurement, providing your lab with the most versatile reader available.
Temperature regulationMicroorganisms have different optimal temperatures for maximum growth rates. To ensure optimal growth conditions, BMG LABTECH readers offer accurate temperature regulation up to 65°C. The specific incubation zone guarantees a uniform temperature and minimizes evaporation. Temperature values are logged to the signal curve, making it simple to compare events at different temperatures.
ShakingMost microbes require aeration to ensure growth or to disperse cultures into an even suspension. Three shaking modes as well as adjustable speed up to 700 rpm or optionally to 1100 rpmprovide optimum settings for your strain. Additionally, the readers can be equipped with an extraordinary robust transport system for shaking 24/7 where required.
Orbital, spiral averaging, and matrix scan
Well scansMicrobial assays are most often non-homogenous. Hence, yourexperiments may benefit from a signal captured over the whole well-surface in either a spiral or orbital pattern, ensuring the most accurate measurement. More detailed matrix scan options (from 2x2 to 30x30) acquire multiple read points over the well surface and display the signal variation in a matrix. This is extremely advantageous when studying highly non-homogenous samples, such as flocculating bacteria or yeast, and for biofilm forming bacteria, to study biofilm size and shape in more detail.
SummaryBacteria communicate by producing, detecting and responding to low molecular mass molecules known as autoinducers (AI) in a process called quorum sensing. The process is well-described in Vibrio fischeri, an organism that turns bioluminescent with high cell density. The molecule responsible for bioluminescence development is N-3-oxo-hexanoyl-L-homoserine lactone (3-oxo-C6-HSL).A BMG LABTECH microplate reader detected biolumine-scence of V. fischeri to report on quorum sensing and relatedit to the cell density determined by OD600 measurements.V. fischeri strains mutated in genes required for lumi-nescence development were exposed to 3-oxo-C6-HSL tostudy cell-to-cell communication. Combining luminescence and absorbance measurements enabled to monitor microbial growth at the same time with bioluminescence. This means that a specific signal and an internal control can be acquired to allow a ratiometric read-out.
Monitoring bacterial cell-to-cell communication “quorum sensing” using a BMG LABTECH microplate readerK.E. Eboigbodin and G.K. Robinson Dept. of Biosciences, University of Kent, Canterbury, CT2 7NJ
3E+4
2.5E+4
2E+4
1.5E+4
1E+4
5E+3
0
RLU
/OD
(600
nm)
0 5 10 15 20 25
Time (h)
KV267-KV240-ESR1-
KV267+KV240+ESR1+
Normalized bioluminescence of V. fi scheri strains. Bacteria were either supplemented with exogenous HSL (+) or not (-). Insert shows growth curves
“After extensive testing of five different brands of microplate readers, only BMG LABTECH instruments were found to have the sensitivity, durability and reliability required for our applications.”
Associate Professor Christopher McDevitt, Group Leader The Peter Doherty Institute for Infection and Immunity,
The University of Melbourne, Australia
Key features to keep both microbes and researchers happy
Atmospheric Control UnitThe living environment of microorganisms is diverse; replicating it in a plate reader requires precise control of the atmosphere. The Atmospheric Control Unit (ACU) independently regulates O2 and CO2 inside the reader to establish an environment conducive for growth. This allows long-term measurements atstable O2 (0.1-20 %) and CO2 (0.1-20 %) atmospheres, perfect forthe most fastidious organisms. Gas ramping can be programmed to study responses to changing gas conditions in real-time.
ACU perfectly regulates both 02 and CO2
Linear, orbital and double-orbital shaking mode
Simplifi ed schematic of the LVF Monochromator technology.
SummaryIt is well known that microorganisms need a specific level of carbon dioxide (CO2) for growth and reproduction. The period during which this level is increased usually corresponds to the lag phase as the organism is unable to divide until the critical concentration of CO2 is reached. Neisseria meningitidis
is considered to require or benefit from a concentration of CO2
greater than atmospheric, hence it is a suitable organism to study CO2 effects.A strain of Neisseria meningitidis was used to assess the efficiency of a BMG LABTECH multi-mode plate reader coupled with an Atmospheric Control Unit (ACU) to deliver 5 % CO2. Growth was determined by detecting absorbance at 405 nm and compared to data obtained on a microplate reader without ACU.The BMG LABTECH plate reader with ACU is able to achieve and maintain a level of CO2 required for such a fastidious organism or for cell cultures.
Growth of Neisseria meningitidis in a BMG LABTECH microplate reader with Atmospheric Control Unit (ACU)Kerry L. Cutter University of the West of England, Bristol
100
90
80
70
60
50
40
30
20
10
0
% B
acte
rial G
row
th
0 5 10 15 20 25
Neat1 in 101 in 1001 in 10001 in 10,0001 in 100,0001 in 1,000,000
Time (hours)
Growth of serially diluted cultures of Neisseria meningitidis in BHI broth supplemented with 10 % FBS using a BMG LABTECH plate reader with ACU set to deliver 5 % CO2 at 37°C. The data presented were calculated from triplicate optical density readings (at 405nm) taken hourly over a 24 h period from duplicate experiments.
For more information please refer to BMG LABTECH application note 155.
For more information please refer to BMG LABTECH application note 199.
Headquarters GermanyBMG LABTECH GmbHAllmendgrün 877799 OrtenbergTel. +49 781 96968 [email protected]
AustraliaBMG LABTECH Pty. Ltd.2/24 Carbine WayMornington, Victoria, 3931Tel. +61 3 5973 [email protected]
FranceBMG LABTECH SARL 7, Rue Roland Martin94500 Champigny s/MarneTel. +33 1 48 86 20 [email protected]
JapanBMG LABTECH JAPAN Ltd.1-6-2, Shimo-choOmiya-ku330-0844 Saitama City Tel. +81 48 647 [email protected]
UK BMG LABTECH Ltd. 8 Bell Business ParkSmeaton CloseAylesburyBucksHP19 8JRTel. +44 1296 [email protected]
USABMG LABTECH Inc.13000 Weston ParkwaySuite 109Cary, NC 27513Tel. +1 877 264 [email protected]
www.bmglabtech.com
04/2
019
Made in Germany
Streamline your bacterial growth assays
SummaryThe bacterium Campylobacter jejuni causes food poisoning with symptoms of abdominal pain, diarrhea and fever. C. jejuni is cultured at 37-42°C, 5-8% O2 and 10% CO2 as this mimics conditions found in the intestine of its hosts.Here, a comparison of C. jejuni growth studies performed in 8 ml scale were compared with growth curves obtained in a 96 well microplate. The atmosphere was controlled with a VAIN workstation or the Atmospheric Control Unit of a micro-plate reader, respectively. Optical density at 600 nm to assess biomass was acquired in a cuvette or directly in the microplate.The comparison of growth of Campylobacter in tubes and in 96 well plates demonstrates that this fastidious organism can be transferred to a 96 well format. This is made easy by the design of the BMG LABTECH microplate reader providing temperature control, shaking and atmospheric control.
Growth of Campylobacter using a microplate reader equipped with ACUR.D. Haigh, J.M. Ketley Department of Genetics, University of Leicester
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
OD 6
00 n
m
0 2 4 6 8 10 12 14 16 18 20 22
Time (hours)
111688117681116
Growth of Campylobacter isolates over a 24 hour period in theFLUOstar® Omega.
Shak
ing
Exte
nsive
sha
king
Mul
ti-m
ode
dete
ctio
n
Incu
batio
n
Wel
l sca
ns
Atm
osph
eric
Con
trol
Uni
t
Wha
t els
e sh
ould
you
kno
w
SPEC
TROs
tar®
Nan
o
Our i
nnov
ative
abs
orba
nce-
only
mic
ropl
ate
read
er h
as th
e fl e
xibi
lity
to p
erfo
rm a
ssay
s qu
ickl
y an
d ea
sily
in m
icro
plat
es o
r via
the
built
in c
uvet
te p
ort.
This
spe
ctro
met
er-
base
d ab
sorb
ance
pla
te re
ader
cap
ture
s a
full
UV/
vis s
pect
rum
in le
ss th
an 1
sec
/wel
l. Its
spe
ed a
nd s
impl
e pu
sh b
utto
n op
erat
ion
mak
e it
the
lead
ing
mic
ropl
ate
read
er fo
r ab
sorb
ance
mea
sure
men
ts.
Omeg
a Se
ries
(Up
to 6
5°C)
The
Omeg
a se
ries
offe
r tru
e fl e
xibi
lity
to
prov
ide
you
with
the
perf
ect r
eade
r to
fulfi
l yo
ur re
quire
men
ts: f
rom
an
abso
rban
ce-
only
SPE
CTRO
star
or l
umin
esce
nce-
only
LU
MIs
tar t
o a
fully
-equ
ippe
d PO
LARs
tar
Omeg
a w
ith u
p to
sev
en d
etec
tion
mod
es.
Your
cho
sen
inst
rum
ent c
an a
lso
be
upgr
aded
at a
ny ti
me
if yo
u ne
ed m
ore
fea-
ture
s or
add
ition
al d
etec
tion
mod
es.
CLAR
IOst
ar®
Plu
s
(Mon
ochr
omat
or)
(Up
to 6
5°C)
The
CLAR
IOst
ar P
lus
is th
e id
eal i
nstr
u-m
ent f
or a
ssay
dev
elop
men
t. Ou
r mos
t fl e
xibl
e m
ulti-
mod
e pl
ate
read
er c
omes
eq
uipp
ed w
ith o
ur p
aten
ted
LVF
Mon
ochr
o-m
ator
s™, fi
lter
s, a
nd s
pect
rom
eter
. Fle
xibi
-lit
y co
mbi
ned
with
the
best
sen
sitiv
ity o
f its
cl
ass,
and
our
new
ly d
evel
oped
Enh
ance
d Dy
nam
ic R
ange
tech
nolo
gy, m
ake
it th
e id
eal r
eade
r for
ass
ay d
evel
opm
ent.
SummaryStreptococcus agalactiae (also known as group B streptococcus [GBS]) is associated with various diseases such as neonatal disease, sepsis, arthritis, pneumonia, meningitis, skin and softtissue infections. Host colonization and virulence are mainly studiedusing microscopy and fluorescent biomarkers. Strategies forlabelling GBS with fluorescent biomarkers have so far been limited to antibody-based immunostaining methods and non-specific protein/DNA stains.Green fluorescent protein (GFP) expression is a common labeling method for bacteria, enabling their identification in complex samples or monitoring of their subcellular locations in eukaryotic host cells. Recently, stable expression of a green fluorescent protein mutant (GFPmut3) with enhanced fluorescence intensityin GBS was reported. Validation and stability of GFPmut3 expression in GBS cultures was measured by the CLARIOstar multi-mode plate reader and its fluorescence intensity, fluore-scence polarization and absorbance measurement capabilities.
Expression of a stable GFP mutant in Group B Streptococcus. Growth, detection and monitoring.Matthew J. Sullivan and Glen C. Ulett, School of Medical Science, and Menzies Health Institute Queensland, Griffith University, Gold Coast, Queensland, Australia
1.00
0.75
0.50
0.25
0.000 3 6 9 12
Abso
rban
ce (O
D 600n
m)
Time (h)
A B3
2
1
0
-10 3 6 9 12
105
RFU
(GFP
FP
inte
nsity
)
Time (h)
Absorbance (A) and polarized fl uorescence intensity profi le (B) of GFP+ GBS (green) and GFP- GBS (black) grown in CDM media.
For more information please refer to BMG LABTECH application note 217.
For more information please refer to BMG LABTECH application note 329.
Headquarters GermanyBMG LABTECH GmbHAllmendgrün 877799 OrtenbergTel. +49 781 96968 [email protected]
AustraliaBMG LABTECH Pty. Ltd.2/24 Carbine WayMornington, Victoria, 3931Tel. +61 3 5973 [email protected]
FranceBMG LABTECH SARL 7, Rue Roland Martin94500 Champigny s/MarneTel. +33 1 48 86 20 [email protected]
JapanBMG LABTECH JAPAN Ltd.1-6-2, Shimo-choOmiya-ku330-0844 Saitama City Tel. +81 48 647 [email protected]
UK BMG LABTECH Ltd. 8 Bell Business ParkSmeaton CloseAylesburyBucksHP19 8JRTel. +44 1296 [email protected]
USABMG LABTECH Inc.13000 Weston ParkwaySuite 109Cary, NC 27513Tel. +1 877 264 [email protected]
www.bmglabtech.com04
/201
9
Made in Germany
Streamline your bacterial growth assays
SummaryThe bacterium Campylobacter jejuni causes food poisoning with symptoms of abdominal pain, diarrhea and fever. C. jejuni is cultured at 37-42°C, 5-8% O2 and 10% CO2 as this mimics conditions found in the intestine of its hosts.Here, a comparison of C. jejuni growth studies performed in 8 ml scale were compared with growth curves obtained in a 96 well microplate. The atmosphere was controlled with a VAIN workstation or the Atmospheric Control Unit of a micro-plate reader, respectively. Optical density at 600 nm to assess biomass was acquired in a cuvette or directly in the microplate.The comparison of growth of Campylobacter in tubes and in 96 well plates demonstrates that this fastidious organism can be transferred to a 96 well format. This is made easy by the design of the BMG LABTECH microplate reader providing temperature control, shaking and atmospheric control.
Growth of Campylobacter using a microplate reader equipped with ACUR.D. Haigh, J.M. Ketley Department of Genetics, University of Leicester
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
OD 6
00 n
m
0 2 4 6 8 10 12 14 16 18 20 22
Time (hours)
111688117681116
Growth of Campylobacter isolates over a 24 hour period in theFLUOstar® Omega.
Shak
ing
Exte
nsive
sha
king
Mul
ti-m
ode
dete
ctio
n
Incu
batio
n
Wel
l sca
ns
Atm
osph
eric
Con
trol
Uni
t
Wha
t els
e sh
ould
you
kno
w
SPEC
TROs
tar®
Nan
o
Our i
nnov
ative
abs
orba
nce-
only
mic
ropl
ate
read
er h
as th
e fl e
xibi
lity
to p
erfo
rm a
ssay
s qu
ickl
y an
d ea
sily
in m
icro
plat
es o
r via
the
built
in c
uvet
te p
ort.
This
spe
ctro
met
er-
base
d ab
sorb
ance
pla
te re
ader
cap
ture
s a
full
UV/
vis s
pect
rum
in le
ss th
an 1
sec
/wel
l. Its
spe
ed a
nd s
impl
e pu
sh b
utto
n op
erat
ion
mak
e it
the
lead
ing
mic
ropl
ate
read
er fo
r ab
sorb
ance
mea
sure
men
ts.
Omeg
a Se
ries
(Up
to 6
5°C)
The
Omeg
a se
ries
offe
r tru
e fl e
xibi
lity
to
prov
ide
you
with
the
perf
ect r
eade
r to
fulfi
l yo
ur re
quire
men
ts: f
rom
an
abso
rban
ce-
only
SPE
CTRO
star
or l
umin
esce
nce-
only
LU
MIs
tar t
o a
fully
-equ
ippe
d PO
LARs
tar
Omeg
a w
ith u
p to
sev
en d
etec
tion
mod
es.
Your
cho
sen
inst
rum
ent c
an a
lso
be
upgr
aded
at a
ny ti
me
if yo
u ne
ed m
ore
fea-
ture
s or
add
ition
al d
etec
tion
mod
es.
CLAR
IOst
ar®
Plu
s
(Mon
ochr
omat
or)
(Up
to 6
5°C)
The
CLAR
IOst
ar P
lus
is th
e id
eal i
nstr
u-m
ent f
or a
ssay
dev
elop
men
t. Ou
r mos
t fl e
xibl
e m
ulti-
mod
e pl
ate
read
er c
omes
eq
uipp
ed w
ith o
ur p
aten
ted
LVF
Mon
ochr
o-m
ator
s™, fi
lter
s, a
nd s
pect
rom
eter
. Fle
xibi
-lit
y co
mbi
ned
with
the
best
sen
sitiv
ity o
f its
cl
ass,
and
our
new
ly d
evel
oped
Enh
ance
d Dy
nam
ic R
ange
tech
nolo
gy, m
ake
it th
e id
eal r
eade
r for
ass
ay d
evel
opm
ent.
SummaryStreptococcus agalactiae (also known as group B streptococcus [GBS]) is associated with various diseases such as neonatal disease, sepsis, arthritis, pneumonia, meningitis, skin and softtissue infections. Host colonization and virulence are mainly studiedusing microscopy and fluorescent biomarkers. Strategies forlabelling GBS with fluorescent biomarkers have so far been limited to antibody-based immunostaining methods and non-specific protein/DNA stains.Green fluorescent protein (GFP) expression is a common labeling method for bacteria, enabling their identification in complex samples or monitoring of their subcellular locations in eukaryotic host cells. Recently, stable expression of a green fluorescent protein mutant (GFPmut3) with enhanced fluorescence intensityin GBS was reported. Validation and stability of GFPmut3 expression in GBS cultures was measured by the CLARIOstar multi-mode plate reader and its fluorescence intensity, fluore-scence polarization and absorbance measurement capabilities.
Expression of a stable GFP mutant in Group B Streptococcus. Growth, detection and monitoring.Matthew J. Sullivan and Glen C. Ulett, School of Medical Science, and Menzies Health Institute Queensland, Griffith University, Gold Coast, Queensland, Australia
1.00
0.75
0.50
0.25
0.000 3 6 9 12
Abso
rban
ce (O
D 600n
m)
Time (h)
A B3
2
1
0
-10 3 6 9 12
105
RFU
(GFP
FP
inte
nsity
)
Time (h)
Absorbance (A) and polarized fl uorescence intensity profi le (B) of GFP+ GBS (green) and GFP- GBS (black) grown in CDM media.
For more information please refer to BMG LABTECH application note 217.
For more information please refer to BMG LABTECH application note 329.