Wednesday, December 26, 2012

Fun fact: Scientist are beginning to understand the molecular basis of why we feel poorly after holiday overeating

There are many reasons why you may wake this morning feeling a bit ill; such as eating rich food and drinking in excess.


Photographic reproduction of 'The Hangover' by Henri de Toulouse Lautrec (circa 1888)

However a recent study shows that there is a molecular basis that can explain why alteration in the amount we eat and the time of day which we eat results in our bodies feeling unwell. A collaboration between UCSF in the U.S. and the Max Planck Institute in Germany entitled 'PKC gamma particpates in food entrainment by regulating BMAL1' sheds new light on the molecular basis of our 'food clock'. The study compared mice with and without PKC gamma and investigated their ability to adjust to a new feeding time. Mice with PKC gamma awoke before their feeding time while mice without slept through the new feeding time.
The concept of the food clock has been accepted as a part of our normal circadian rhythms which allows us and other animals to make the most of our available food resources. The food clock initiates changes that anticipate feeding time so that our bodies can best utilize food when it arrives. This helps to explain why you are ready for lunch at the same time each day or the birds at your birdfeeder arrive at the same time every morning. While this clock can be reset overtime, overeating and eating at odd times may lead to disruptions at the molecular level that affect your clock and therefore how you feel the day after.

Tuesday, December 25, 2012

Merry Christmas

Merry Christmas
                Froehliche Weihnachten
                                               Feliz Navidad
                                                          Joyeux Noel
                                                                       Mele Kalikimaka
                                                                                           Boas Festas
                                                                                                               etc.
From BMG LABTECH

Description
Deutsch: Christmas goose (Weihnachtsgans)
DateDecember 2004(2004-12)
SourceOwn work
AuthorJürgen Howaldt

Monday, December 24, 2012

Happy Christmas Eve

BMG would like to wish you all a happy Christmas Eve. If you are traveling we hope you arrive at your destination safely.
 
DescriptionThis was another of the competitors at the St. Croix Valley Carriage and Driving Society's 2008 Sleigh and Cutter Festival held in Lake Elmo, MN. This portland cutter is being pulled by a a Norwegian Fjord.
Date27 January 2008(2008-01-27), 11:59
SourceUphill Sleigh Ride
AuthorPete Markham from Loretto, USA

Friday, December 21, 2012

Focus on Prion Assay



Prions are known to cause various neurodegenerative disorders or transmissible spongiform encephalopathies (TSEs) in animals and humans. They cause an abnormal folding of prion proteins mostly found in the brain, so when transmitted the disease causes brain damage and usually death. Some prion diseases include Scrapie in sheep; Chronic Wasting Disease (CWD) in deer; Bovine Spongiform Encephalopathy (BSE) in cows; and Creutzfeldt-Jakob Disease (CJD) in humans. In order to better understand these diseases the levels of prions present in an infection needs to be measured. A new assay, Real-Time Quaking Induced Conversion Assay (RT-QuIC), developed at Rocky Mountain Laboratories on a BMG LABTECH Omega plate reader is faster and higher throughput than previous bioassays using entire animals as hosts. Now the assay can be performed in a 96 well plate format in a few hours to a couple of days compared to several months or a year. For more information, see our application note .
 
RT-QuIC end-point dilution analysis of three 263K-inoculated preclinical 10 days post injection hamster BHs. In this case, the approximate SD50 was achieved with a 2 μl aliquot (the seed volume) of a 10E−5 dilution of the scrapie 10dpi 263K BH stock (green line). This gave an SD50/2μL of 10E5.5 and an SD50/g of 10E8.2. This figure was adapted from Rapid End-Point Quantitation of Prion Seeding Activity with Sensitivity Comparable to Bioassays
 

Thursday, December 20, 2012

Applications Thursday


Researchers at Rocky Mountain Laboratories in Hamilton, MT with collaborating scientists in Japan have developed a new assay for quantitating prions for neurodegenerative disease research in transmissible spongiform encephalopathies like Scrapie and Creutzfeldt-Jakob Disease. This new assay, Real-Time Quaking Induced Conversion Assay (RT-QuIC) is faster, of a higher throughput, and more sensitive as compared to previous test methods using biological tissue samples from infected animals. This assay can be measured using BMG LABTECH’s Omega series of  microplate readers. With the Omega’s powerful control and analysis software, researchers can easily perform this assay in several hours to a few days.

For more information please see their publication Rapid End-Point Quantitation of Prion Seeding Activity with Sensitivity Comparable to Bioassays in PLoS Pathog 6(12): e1001217. Further information can also be found on our customer focus page and on an application note  detailing how to run the assay on an Omega microplate reader.

Figure legend: Magnified 100X, and stained with H&E (hematoxylin and eosin) staining technique, this light photomicrograph of brain tissue reveals the presence of prominent spongiotic changes in the cortex, and loss of neurons in a case of variant Creutzfeldt-Jakob disease (vCJD). This image is courtesy of CDC/Teresa Hammett. Photo Credit: Sherif Zaki; MD; PhD; Wun-Ju Shieh; MD; PhD; MPH
 

 
 

Wednesday, December 19, 2012

Fun Fact: December 19, 2012

Albert A. Michelson, winner of the 1907 Nobel prize in Physics was born on this day in 1852. According to nobleprize.org he won the award 'for his optical precision instruments and the spectroscopic and metrological investigations carried out with their aid'. Michelson is best known for his attempts to measure the speed of light. His final tests, completed after his death, resulted in a value of 299,774 km/s. Although subsequent tests have more precisely determined the speed of light his measurments and determination that the speed of light is a constant were among his important contributions.
 


 

Tuesday, December 18, 2012

Did you know that BMG LABTECH instruments have been cited in over 3,000 articles?

As a testament to our long standing history in the industry and to our quality engineered products, BMG LABTECH microplate readers have been cited in over 3200 publications, application notes, and posters. Versatile, robust, and easy to use, BMG LABTECH microplate readers can perform the most simple absorbance assay as well as the most complex advanced cell-based assay.

Find out more about the many applications in our Application Center. You can also Search Our Application Section or you can Search Our Blog for any relevant applications that may have been published or discussed. 

BMG LABTECH - Engineering a better biological solution. 


Monday, December 17, 2012

FAQ: Will BMG LABTECH attend SLAS2013?



Yes, BMG LABTECH is a Diamond Sponsor of SLAS2013 in Orlando Florida. Visit BMG LABTECH at Booth #721 to learn how a perfectly engineered instrument can give a better biological solution and to find out how to win a free iPad Mini.

It is no more apparent how quality engineering can enhance biological applications than with the PHERAstar FS. The PHERAstar FS has become the Gold-Standard Microplate Reader for high-throughput screening labs and core-facilities, quickly replacing outdated, less sensitive measurement instrumentation.

The PHERAstar FS will be highlighted at several workshops, posters, and at different collaborators’ booths at SLAS 2013.

Workshops - refreshments will be served:

Monday 12:30 – 1:45, Naples Room 2-3
  • Labcyte-Miniaturized and Automated Cisbio HTRF® Assays with the Echo® Liquid Handler and BMG PHERAstar FS
Tuesday 12:30-1:15, Sun Room 1-2
  • BMG Labtech-Enhanced Assay Development and High-Throughput Screening with New and Unique Microplate Based Technology. 
Posters:
  • Quantifying Fluorescent Ligand Binding to GPCR’s in Live Cells using the PHERAstar FS – a new format for HTS 
  • Nano High-Throughput Screening (nHTS) Platform-Miniaturization of Cell-based GPCR and Kinase Assays in 3456-well Microplates 
  • Methyltransferase, Acetyltransferase, Kinases, and GTPases Can All Be Measured with Transcreener® Assays and the PHERAstar 
  • Monitoring Drug Solubility and the Growth of Candida albicans Using BMG LABTECH’s NEPHELOstar Light-Scattering Microplate Reader 
  • A new HTRF® assay for the quantification of active Glucagon-like Peptide-1 (GLP-1) (CISBIO poster) 
  • High-Throughput Homogeneous Histone H3 Methyltransferase (HMT) and Demethylase (HDMT) Enzyme Assays using HTRF Technology: G9a, MLL1, and EZH2 Methyltransferase plus LSD1, JMJD2C and JMJD2A Demethylase Assays (CISBIO poster).

Friday, December 14, 2012

Focus On: SNP Genotyping Using the Amplifluor® System and the FLUOstar




SNPs (Single nucleotide polymorphisms) are places in genomic DNA with a single nucleotide difference between individuals in a species. SNPs are a common occurring mutation found in DNA sequences. These genetic variations can occur in coding or non-coding regions and can either have an extreme or no effect on a particular organism. Therefore SNP detection has become important for medical research and pharmacology.

The detection of SNPs (SNP Genotyping) can be achieved with a number of different methods. In application note 187 a homogeneous fluorescence based method on PCR is described. The Amplifluor® SNPs HT Genotyping System from Millipore was used to screen a number of samples with the help of a FLUOstar from BMG LABTECH. 

See application note 187  to learn more about this SNP assay or go to Millipore's website see a more detailed description of this assay.

SNP Graph
A typical pattern of SNP samples. The blue dot in Q1 is buffer. The red dot in Q2 shows the mutant control (green allele), Yellow dots in Q4 are wild type control(red allele) and the orange dot in Q3 represents the heterozygote control(red green allele). Figure 3 clearly shows that all controls that are homozygous for either allele or heterozygous or a mixture only are clearly separated. In addition, samples are precisely located in their clusters.

Thursday, December 13, 2012

High Speed FRET based SNP Genotyping Measurement on the PHERAstar

Automated solution for fast SNP genotyping
  • Optimized optical modules with dual emission
  • More accurate results with ratiometric measurements
Single Nucleotide Polymorphisms (SNP`s) have become an invaluable tool in the field of Genetic Research. Here we show the use of BMG LABTECH´s PHERAstar multimode HTS plate reader for high speed FRET based SNP Genotyping measurement. The new KASPar™ system, developed by KBiosciences, was used to assess the best performance of the PHERAstar. Optimized optical modules with Dual Emission lead to improvements in speed and accuracy of measurement. Can also use with Taqman® and Invader SNP genotyping chemistries.

To learn more see the application note here...

transcreener

Wednesday, December 12, 2012

Did you know that BMG LABTECH has a new webinar on YouTube called Optimizing Cell-Based Microplate Assays?

BMG LABTECH, in collaboration with Labcyte and MRC-T, presented a webinar on Drug Discovery and Development that detailed different ways to optimize cell-based microplate assays. The three speakers and titles were:
  • Optimizing Measurements for Microplate Cell-based Assays
    Catherine Wark, BMG LABTECH, UK
  • Miniaturization and Automation of HTRF® Cell-based Assays with the Echo® Liquid Handler
    Bonnie Edwards, Labcyte, Inc., USA
  • Identification and characterization of allosteric modulators of GPCRs using an HTRF® cellular assay
    Jeff Jerman, PhD, MRC Technology, UK




To see a replay of this webinar go to BMG LABTECH's YouTube page and click on the video "Optimizing Cell-Based Microlate Assays."

Cell Based Assays with the PHERAstar FS


Fun Fact Wednesday – To RT-PCR or Not To RT-PCR

Most labs have access to a thermocyler or PCR machine that can cycle 96 samples in an hour for SNP genotyping.  If you want to genotype a freezer full of samples using allele specific primers, you could analyze about 96 an hour, or 768 in an eight hour day.  If you wanted to collect data more quickly, you could add more and more thermocyclers and have them running in parallel…or you could detect your genotypes on a plate reader.

Method example for GWA study designs
Using water-bath batch cyclers, it is possible for you to cycle hundreds of plates at a time—and with 384-samples per plate, not just 96 samples at a time.  The PHERAstar FS can read a 384-well SNP plate in 28 seconds (two measurements). That means that one hundred and twenty plates an hour can be measured, for eight hours, at 384-samples per plate is 386,000 samples per day—480x more than the original setup.  If you add multiple PHERAstars onto your robotic plate handler, you could genotype about 1.2 million samples a day.  If you switched to 1536-well plates, you could collect genotype data from 2.4 million samples in eight hours! This also does not take into account that the misread rate is much smaller on the PHERAstar FS than on an RT-PCR instrument, further saving time and money. 

Friday, December 7, 2012

Focus On: Dual Emission Detection for FP, FRET, TR-FRET, BRET, and Dual Glow Assays

Dual Emission Detection is a feature in BMG LABTECH microplate readers that allows them to measure two emission wavelengths with only one read of the microplate. This greatly benefits assays like Fluorescence Polarization (FP), Forster resonance energy transfer (FRET), Time-Resolved-FRET (TR-FRET), Bioluminescence resonance energy transfer (BRET), and Dual Glow assays

The benefits include faster measurement times (twice as fast) because it only has to read the plate once. But more importantly it eliminates read-to-read variations that can occur, such as different well volumes, concentrations, meniscus effects, or fluctuations in excitation energy. The result is an assay that is faster, is more sensitive, has a higher Z' factor, and has a lower %CVs.

Dual Emission Detection is found on the PHERAstar series of HTS microplate readers, as well as on the POLARstar Omega and POLARstar OPTIMA.

Dual Emission in the PHERAstar

Thursday, December 6, 2012

Applications Thursday: Simple Apoptosis Assay in an Absorbance Microplate Reader


Apoptosis is a multistage process during which activity of caspase enzymes fluctuates, DNA becomes fragmented and phosphatidyl serine is transferred to the outside of the cell membrane. Apoptosis is a common pathway that can go wrong in many cancers and it is a highly researched area for different cell types. Common methods for apoptosis analysis include:
  • Caspase activity assay, either colorimetric, fluorescent, luminescent or antibody based
  • TUNEL assay, based on DNA fragmentation.
  • Annexin-V assay, based on the binding of dye or fluorescently-conjugated Annexin-V to phosphatidyl serine which has translocated to the cell membrane exterior during apoptosis.
A simple and easy-to-use apoptosis assay, Bicolor's APOPercentage Apoptosis Assay™, was performed on BMG LABTECH's SPECTROstar Nano microplate reader. This assay format takes advantage of the phosphatidyl serine (PS) that is found on the exterior cell surface when apoptosis begins to occur. The APOPercentage dye, which binds to PS, is then taken up by cells undergoing apoptosis. The dye can be quantified by an absorbance instrument such as the SPECTROstar Nano


Table 2
Colorimetric Quantification. Graph Showing Effect of Hydrogen Peroxide (0 – 10mM) on CHO Cells. Results expressed as mean absorbance for triplicate wells ± S.E.M. (n = 3). Exposure time to H2O2 was 4 hours. 

Tuesday, December 4, 2012

Did you know that there are eight different detection modes in BMG LABTECH microplate readers?

Microplate readers have evolved from simple single-mode instruments that can only do a handful of applications in one mode, to complex multimode instruments that can measure thousands of different assays in several modes. Most applications use the three most common detection modes of absorbance, fluorescence intensity, and luminescence and these are the three most common detection modes found multimode microplate readers. Whereas higher end multimode instrumentation can perform applications that are based on specialized chemistry using other detection modes such as AlphaScreen®, Fluorescence Polarization, Time-Resolved Fluorescence (TRF)TR-FRET/HTRF®, and Nephelometry

Learn more about BMG LABTECH's multimode microplate readers and learn more about the different detection modes here

Light-Scattering or nephelometry is a specialized detection mode that can be used
to measure drug solubility in the NEPHELOstar Plus microplate reader. 


Thursday, November 29, 2012

Applications Thursday – GPCR Activation via cAMP or Inositol Phosphate Production is Measured in Live Cells Using HTRF® Chemistry


GPCRs have two major signaling pathways - the regulation of cAMP levels and the increase in intracellular Ca2+ triggered by inositol 1, 4, 5- triphosphate (IP3). Specific G proteins activates these signaling pathways and they are associated with different receptors. Activation of a Gs or Gi coupled receptor results in the increase or decrease of cAMP levels, respectively. While activation of a Gq coupled receptor activates phospholipase C (PLC) and triggers the inositol phosphate (IP) cascade.

Cisbio Bioassays has assay kits able to measure the activation of Gs, Gi and Gq coupled receptors in one assay, using their new generation Lumi4-Tb™ TR-FRET Cryptate. This chemistry allows for the detection of two signals in one well using a green acceptor (λ = 520 nm) and a red acceptor (λ = 665 nm). This same chemistry is used in Cisbio’s Tag-lite® technology.

IP-One and cAMP experiments were performed on the PHERAstar HTS microplate readers using Simultaneous Dual Emission detection (detailed in Application note 209 . With this unique feature, the plate is read only once for dual emission assays, thereby decreasing time and variability. All Cisbio chemistry can be performed on the PHERAstar FS, which uses a UV Laser for excitation. The UV Laser will allow for greater sensitivity and faster read times (see Application Note 222 for IP-One data using the UV Laser) . The POLARstar and FLUOstar Omega with advanced assay technology also perform this chemistry, but in a non-HTS fashion.

Wednesday, November 28, 2012

Fun Fact: One Week Left to Vote for SLAS Board of Directors


The Society for Laboratory Automation and Screening (SLAS) is electing three new members to serve for three years on the SLAS Board of Directors and there is only one week left to vote. Amongst the 8 candidates is Dr. E.J. Dell, the International Marketing Manager for BMG LABTECH. Dr. Dell has attended both ALA and SBS meetings over the last six years while with BMG LABTECH and he hopes to bring some fresh ideas to SLAS if chosen. Some of those ideas include:

  • Further miniaturization to smaller volumes – This will not only save time and money, but assays will also be performed in a more realistic lower volume environment 
  • Increased presence of core-facilities at SLAS – Core facilities that do specialized screening have changed the landscape of HTS 
  • Broadening automation and screening to other scientific fields like Toxicology and Environmental Biology – This would encourage those fields of study to migrate away from whole animal bioassays

If you are a member of SLAS and have not yet voted, you may do so here: https://www.slas.org/election/index-2013.cfm?goback=%2Egmp_3363923%2Egde_3363923_member_189718058.

See you in January at Booth #721 at SLAS2013 in Orlando!

Tuesday, November 27, 2012

Did you know that BMG LABTECH is presenting a webinar tomorrow, November 28th at 12:00 EST on Cell-Based Microplate Assays?


In collaboration with Drug Discovery and Development, BMG LABTECH is hosting a free webinar entitled Developing and Optimizing Cell-Based Microplate Assays. In this webinar, three different speakers will highlight different aspects of developing cell-based assays in a microplate format. They include:

  • Optimizing Measurements for Microplate Cell-based Assays
    Catherine Wark, BMG LABTECH, UK
  • Miniaturization and Automation of HTRF® Cell-based Assays with the Echo® Liquid Handler
    Bonnie Edwards, Labcyte, Inc., USA
  • Identification and characterization of allosteric modulators of GPCRs using an HTRF® cellular assay
    Jeff Jerman, PhD, MRC Technology, UK

Sign up here to join us at 12:00 EST tomorrow for this live webinar, panelists will be available for questions. Reproductions of the webinar will be available shortly upon request.

Monday, November 26, 2012

FAQ: What are the advantages of well scanning?


Under normal reading conditions, most plate readers take a single measurement in the center of each well. For cell based assays where the cell distribution is uneven a single center well reading is insufficient. In some cases, a single center point reading may completely miss the material of interest in a large well format.

To overcome this problem, BMG LABTECH has added an advanced Well Scanning feature to its plate reader lines. In Well Scanning mode the microplate reader can take multiple measurements in each well with up to 30 x 30 data points. Well scanning (30 x 30 matrix) and Direct Optic Bottom reading provide the PHERAstar FS with an excellent platform for advanced high resolution cell-based analysis in 384 well plates. This enables the PHERAstar FS to perform cell-based assays that other microplate readers cannot.


Friday, November 23, 2012

Focus On: Developing and Optimizing Cell-based Microplate Assays Webinar


On Wednesday November 28th at 12:00 EST, Drug Discovery and Development will host a webinar that details how to develop and optimize cell-based assays in a microplate format.

Many researchers need to migrate a biochemical assay to a cell-based format to get more information, or they need to increase their throughput of their current cell-based assay. One way to achieve this is to create a cell-based assay in a microplate format. Over the last several years many new cell-based assays have been developed, in addition, there have been improvements in instrumentation that are needed to measure and dispense these cell-based assays. Join this webinar to hear three presenters discuss three different aspects of cell-based microplate assays:


  • Optimizing Measurements for Microplate Cell-based Assays
    Catherine Wark, BMG LABTECH, UK
  • Miniaturization and Automation of HTRF® Cell-based Assays with the Echo® Liquid Handler
    Bonnie Edwards, Labcyte, Inc., USA
  • Identification and characterization of allosteric modulators of GPCRs using an HTRF® cellular assay
    Jeff Jerman, PhD, MRC Technology, UK


For more information please visit our webinar sign-up page. The webinar is free to join so sign up now!

Thursday, November 22, 2012

Applications Thursday – Dual Glow Reporter Gene Assays Benefit from Dual Emission Detection and Longer Bandwidth Filters

A new second wave of dual glow luciferase reporter gene assays are now available from various companies (Pierce, Promega, Life Technologies). Other than Firefly and Renilla, there are now Gaussia, Cypridina, Green Renilla, and Red Firefly luciferases that are available and that can be multiplexed together, or even with fluorescent signals.

One main difference between these new dual glow reporter assays  and earlier dual glow assays like DLR™, is that the two different luminescent signals can be differentiated at different wavelengths, rather than at different time points. This enables higher throughput because there is no need for injection and no need for a minimal measurement time, the only limitation is in the instrumentation.

Larger bandwidth filters can capture more signal. 
Two features on instruments that will benefit these dual glow assays are Dual Emission Detection, and Longer Bandwidth Filters. Since two emission wavelengths need to be measured per well, microplate readers with Dual Emission Detection, like the POLARstar Omega and PHERAstar FS, will measure these assays in half the time compared to instruments that do not have Dual Emission Detection. Dual Emission Detection also helps to correct with read-to-read variations that can occur with reading the plate twice.

In addition, since these luminescent signals are not as bright as fluorescent signals, it benefits from collecting a larger area of the emission signal at each wavelength. Thus microplate readers with the ability to measure broad bandwidths (50-100 nm), like the POLARstar Omega and PHERAstar FS because they use longer bandwidth filters, will be more sensitive in these assays.

Learn more about it here, Wavelength Based Dual Glow Reporter Genes.

Wednesday, November 21, 2012

Fun Fact – Longer Telomeres, Mean Longer Life Expectancy

Warbler A new 20 year study published in Molecular Ecology shows that telomere length can be used to predict the lifespan of Seychelles Warblers, which are birds contained on an island and that have no natural predator. This study measured telomeres, which are end caps of chromosomal DNA that get shorter each time DNA is copied. Across the lifespan of these warblers, the study found that having shorter telomeres at any age is associated with an increased risk of death. The study showed that “short and rapidly shortening telomeres were a good indication that the bird would die within a year.” Since telomeres can be attacked by oxidants, it is thought that stress may play a role in telomere shortening. Though this is just one study, it seems that telomere length is a better indicator of biological age and future life-expectancy than real age.

For more information click here: http://www.sciencedaily.com/releases/2012/11/121119213144.htm

Tuesday, November 20, 2012

Did you know that a BMG LABTECH microplate reader can take some of the workload off of your High Content Screening (HCS) system?

High content screening allows researchers to see the phenotype of a cell and how it is affected by various treatments. Information about cell shape or where two fluorescent proteins co-localize isn’t information that can be obtained many other ways. However, some changes—cell motility, cell death, fluorescent protein expression levels—might require more information than a standard plate reader can provide but don’t require the full imaging capability of an HCS system.

Readers like BMG LABTECH’s PHERAstar FS can focus precisely from the bottom of the well and measure cell-based assays in scanning mode, taking hundreds of data points per well to create a fluorescence intensity map. It isn’t possible to see what’s happening in individual cells, but it’s easy to see cell coverage and uniformity, or where in the well expression is happening. An assay that can take two hours on an HCS system would take only eight minutes on the PHERAstar FS, meaning that researchers can collect more data per hour and that expensive HCS equipment is free to be used for methods that truly require its capabilities.
This figure shows the well scanning capabilities of the PHERAstar FS
using a 384-well plate seeded with cells expressing different levels of GFP.
 Here is an application note that describes in detail how the PHERAstar FS outperforms a leading CCD device in HTRF® mode:
HTS Instrument Discovers Low Affinity Inhibitors of the Inositol Phosphate (IP) Signaling Pathway.

For more information on Developing and Optimizing Cell-based Microplate Assays, join us for our webinar on Wednesday, November 28. More information can be found here:
http://www.bmglabtech.com/webinar/cell-based-assays.cfm

Monday, November 19, 2012

FAQ: How can cell migration or invasion assays be measured on a microplate reader?

Cell migration and cell invasion are important parameters to investigate for different cell types and for different disease states. For instance, tumor cells undergo a transformation where they metastasize to another area of the body by invading different tissues. Or leukocytes need to migrate to different areas of the body during infection. Therefore, assays that can measure cell migration and cell invasion are important tools to have for a microplate reader.

One way to measure cell invasion is to use the FluoroBlokTM assay from BD. Well chambers with a membrane are placed into a 96 well microplate and fluorescently labeled cells are seeded on top of the membrane. Measurements are taken from the microplate bottom to measure how quickly the fluorescently labeled cells invade through the membrane. Review this application note to learn more: Analysis of Prostate Tumour Cell Invasion Using BD FluoroBlok™and FLUOstar OPTIMA

Another way to measure cell migration is to use the Platypus® OrisTM Cell Migration Assay. Here a plug is placed in the center of a microplate well and fluorescently labeled cells are seeded on the outside of the plug. The plug is the removed and measurements are taken to watch how quickly the migration of the cells occurs toward the middle of the well. Review this application note to learn more: Analysis of migration using the Oris(TM) Cell Migration Assay-TriCoated kit on the POLARstar Omega.

Friday, November 16, 2012

Focus On Histone Deacetylases (HDACs)

Complete Histone with DNAAs the field of epigenetics unfolds, various enzymes have now become important targets for drug screening as cures to various diseases, such as cancer. One such enzyme family is the Histone Deacetylases (HDACs), which modify histones and which in turn helps to regulate gene transcription. Though the mechanism of action is not fully known, it has been shown that inhibition of HDACs results in the overacetylation of histones that in turn can lead to a controlled cell death or apoptosis. As such, several HDAC inhibitors (HDIs) are in phase I or II clinical trials, for example suberoylanilide hydroxamic acid (SAHA; ZOLINZA®, Merck).

Check out one of our POLARstar application notes that describes an HDAC assay: A Fluorescence Based Assay of the Epigenetic Enzyme Histonedeacetylase 1 (HDAC1)

Thursday, November 15, 2012

Inside Applications: Methyltransferase, Acetyltransferase, Kinases, and GTPases

Methyltransferase, Acetyltransferase, Kinases, and GTPases Can All Be Measured with Transcreener® Assays

  • The universal Transcreener® assays measure the byproducts of common biological enzymatic reactions
  • For epigenetic studies, AMP/GMP can be monitored for methyltranferase or acetyltransferase activity
  • ADP for kinases, ATPases, or helicases; GDP for Gproteins; and UDP for glycosyltransferases
This application note demonstrates the validation of the BMG LABTECH PHERAstar Plus and PHERAstar FS instruments for use with Transcreener FP assays in 384-well format. BellBrook Labs offers four competitive immunoassays for direct detection of ADP, AMP/GMP, UDP and GDP. These nucleotides are often byproducts in enzymatic reactions and their quantification allows calculation of enzymatic activity.

Transcreener is a registered trademark of BellBrook Labs.

Wednesday, November 14, 2012

Fun Fact: Right now, Epigenetic is one of the fastest growing fields of life science.

The field of Epigenetics has risen in popularity over years, as such so has the number of publications with the term Epigenetic in the title or abstract. Starting in 1990, the year and the term Epigenetic was searched on Pubmed  to see the number of results that were returned. Though this graph is a crude estimation, if nothing else it shows the growing trend for using the term epigenetics in research publications. It is compared to the term Proteomics which also had a rise in use but seemed to peak earlier in the decade. Will Epigenetics also peak ~1000 publications/year?

Tuesday, November 13, 2012

Did you know about the different epigenetic assays that can be performed on a microplate reader?


Multimode microplate readers have become a necessary tool in the modern life science laboratory and as a consequence many biochemical and cellular assays are made specifically for a microplate format. With research into epigenetics exploding over the last couple of years, the number of assays available for microplate readers has also grown.

Here are some different companies that offere assays that can all be performed on a BMG LABTCH microplate reader:


DNA methylation



Monday, November 12, 2012

FAQ: What is epigenetics and why is it such an exciting field of biology right now?


Epigenetics  is the study of physical changes to the genome that influence gene expression and that can lead to inheritable changes in cells. Basically epigenetics put to rest the argument of nature vs. nurture, it is both.

Specifically epigenetics encompass three changes to the genome:

  • DNA methylation is thought to be involved in silencing regions of DNA so they cannot be transcribed
  • Histone modification (i.e. acetylation, methylation, ubiquitylation, phosphorylation and sumoylation) can lead to histone remodeling, possibly making it easier for transcription 
  • Small Interfering RNA (siRNA) are small fragments of RNA (<30 nucleotides) which can bind to and regulate different promoter regions of DNA. 


The study of epigenetics has led to important understandings of  genetic differences, such as why one identical twin can contract a disease such as lupus, while the other one does not. In addition, epigenetics is allowing us to change the way we view and treat some cancers such as squamous cell carcinoma and liver cancer.

As a consequence new assays are being developed all of the time to study epigenetics. Revisit our Blog in the coming week as we will investigate some of those assays that can be performed on a microplate reader.
Epigenetic mechanisms

Friday, November 9, 2012

Friday Focus: Luciferase Reporter Assays


Reporter genes like luciferase are one of the most fundamental and useful tools in a molecular biologist’s tool box. As popular and as useful as something like Firefly Luciferase is, it isn’t ideal for every application. At 61kDa, it’s large enough that as a fusion protein, it may interfere with some of the processes it’s intended to track. Luciferase substrate, D-luciferin, produces detectable levels of background luminescence that interfere with detecting biologically relevant signals from the reporter itself. And any luciferase reporter would be more useful if it could be detected at lower levels.

Firefly LuciferasePromega’s new NanoLuc LuciferaseTM reporter (isolated from a deep sea shrimp) is an enhanced tool for researchers. It’s more than three times smaller than Firefly Luciferase, so it’s less likely to interfere with biological processes. Nanoluc’s substrate produces lower levels of background luminescence than traditional luciferase assays and the NanLuc enzyme itself emits light about 100x brighter than Firefly. This means that researchers are more likely to see low levels of activity and also design assays that work at biologically relevant concentrations. Thousands of BMG LABTECH customers use Promega’s reporter genes on our microplate readers  daily and we know they are excited to try the newest one out too.

Thursday, November 8, 2012

Applications Thursday – Chroma-Glo Assay in 1536-well Microplates



Screening facilities require simple solutions for measuring and correlating a variety of parameters of cellular processes. Robust assays and high-precision, low-volume, non-contact pipetting systems are now providing this capability. This application highlights the scalability of the CellTiter-Glo®, Caspase-Glo™ 3/7, and Chroma-Luc™/Chroma-Glo™ Technologies for high-throughput and ultrahigh-throughput cell-based screening in low-volume 384- and 1536-well formats.



For more information click here

Wednesday, November 7, 2012

Fun Fact: Movember

According to uk.movember.com:
"Movember, the month formerly known as November, is when brave and selfless men around the world grow a moustache, with the support of the women in their lives, to raise awareness and funds for men’s health - specifically prostate and testicular cancer.

Movember was established in 2003 by a few friends over a beer in a pub just outside Melbourne, Australia. The goal was simple – to create a campaign promoting the growth of the moustache among likeminded people and having fun along the way. It is about real men, talking about real issues and changing the face of men’s health, one moustache at a time. Movember now spans the globe, with campaigns in 21 countries in 2012."

Four members of the BMG LABTECH UK team are proud to be paritcipating and raising money and awareness for men's health issues.

MOUSTACHECheck up on the BMG LABTECH team including:

Robert Mount
Martin Lane
Lee Archer
Martin Fisher

and help us raise money and awareness for this cause.

For more information visit: http://uk.movember.com/

Tuesday, November 6, 2012

Did you know that you no longer need injectors to perform dual reporter assays using luciferase?

Photinus pyralis Firefly glowingStable and bright luminescence reporters like red-luciferase, green-Renilla, Gaussia and Cypridina lets users measure two luciferase constructs at once using a long-lived luminescence suitable for liquid handling. Traditional dual reporter assays, like dual luciferase, use time and two different injections to allow users to discriminate between both reporters. Glow-type dual reporter assays use two luciferase constructs that each emit a light at a different wavelength, meaning that a luminometer capable of accepting filters, like any of BMG LABTECH’s Omega microplate readers, can discriminate between two different luciferase enzymes emitting at the same time.

For more information about some of these newer luminescence assays please see Pierce or Promega.

Friday, November 2, 2012

Focus On BMG LABTECH Customers

For over 20 years BMG LABTECH has been providing quality instrumentation for life science labs, core facilities, and high-throughput screening centers. As such we have tens of thousands of satisfied customers. If you are looking for a new microplate reader and want an unbiased opinion, read and listen to what our customers have to say about BMG LABTECH products and service. German engineering is just the beginning, satisfied customers are the result.

"The Cancer Research UK Drug Discovery Unit at the Paterson Institute for Cancer Research has recently purchased a PHERAstar FS multimode plate reader. I had previously used this plate reader and worked with BMG over a number of years.  I have always been very impressed with the quality of data produced by the PHERAstar across various applications. The customer service, from purchase right through to downstream support, has always been prompt, friendly and incredibly helpful. Our lab was in need of a top spec plate reader that would produce the highest quality data for our assay scientists yet remain easy to use and approachable for occasional users. When demonstrated alongside other top spec plate readers, the PHERAstar outperformed on each level, with the  most marked improvements seen when measuring europium-based TR-FRET."
Alexandra Boakes
University of Manchester, UK
The Paterson Institute for Cancer Research, Drug Discovery Unit


Read more customer testimonials here: http://www.bmglabtech.com/testimonials/testimonials.cfm.

Thursday, November 1, 2012

Applications Thursday – SNP Genotyping

Single Nucleotide Polymorphisms (SNPs) can be used as genetic markers in Agrigenomics to identify seeds or plants in breeding programs. There are several different chemistries that can perform SNP Genotyping amongst which are KASPar®, Taqman®, and Invader®. These chemistries rely on the detection of multiple fluorophores for each sample and when screening thousands of samples an instrument is needed that can reliably read these multiple fluorophores in a high-throughput manner.

 The PHERAstar Plus and the PHERAstar FS are the perfect instruments to handle these screens. Having Dual Emission Detection and the ability to read 1536-well microplates in less than a minute, the PHERAstar microplate readers make SNP Genotyping a snap.

For more information, check out this application note showing how the misread rate of SNP Genotyping using KASPar® is less than 0.1, while a recent publication shows how the PHERAstar and KASPar® have been used for the genetic mapping of Pigeonpea and Legumes.

Tuesday, October 30, 2012

Did you know that there is now a German website available?

Herzlich Willkommen auf der Website von BMG LABTECH !

It is now possible to find out more about the products, applications, news and more in German on BMG LABTECH's website. Visit http://www.bmglabtech.com/deutsch to get the latest information on our microplate readers in German.

The addition of German language pages to the website brings the total number of languages available to 8 - including Japanese, French, Portuguese, Spanish, Italian, Swedish, and of course English.

Monday, October 29, 2012

FAQ: What is the difference between a spectrometer and a monochromator for absorbance measurements?

A spectrometer incorporates a highly efficient optical grating and a solid state array detector that allows the measurement of light intensity throughout the entire UV and visible parts of the spectrum. Similar to a monochromator, but much faster, the spectrometer allows you to capture the whole UV/Vis spectrum (220-1000 nm) of a sample within one second per well – no scanning is needed.

In practical terms, a monochromator only captures one measurement in the UV-Visible spectrum at a particular wavelength or a particular bandwidth, and therefore gives only one value. Whereas a spectrometer captures the entire UV-Visible spectrum in the same amount of time, giving values at every wavelength.

Here is a graphical representation of this – notice all of the information that is lost if a monochromator is used:

Find out more about this technology in an article on Genetic and Engineering News:
Microplate Reader Absorbance Assays - New Tools Bridge Gap between Single and Multiple Sample Absorbance Instruments.
For more information on the UV/Vis absorbance spectrometer technology used in BMG LABTECH's instruments, visit http://www.bmglabtech.com/technology/spectrometer.cfm/. BMG LABTECH instruments that have a spectrometer include: SPECTROstar Nano, Omega Series of readers, and the PHERAstar FS.

Thursday, October 25, 2012

Applications Thursday: RT-QuIC, a Faster, Higher Throughput Assay to Measure Prion Diseases

Aphis.usda.gov BSE 5Researchers at Rocky Mountain Laboratories in Hamilton, MT with collaborating scientists in Japan have developed a new assay for quantitating prions for neurodegenerative disease research. This new assay, Real-Time Quaking Induced Conversion Assay (RT-QuIC) is a faster, higher throughput, and more sensitive assay for prions than compared to previous test methods using biological tissue samples from infected animals.

BOVINE PRION PROTEIN 1dx0 asym r 500This new prion assay can be measured using BMG LABTECH’s Omega series of microplate readers . With the Omega’s powerful control and  MARS Data Analysis Software, researchers can easily perform this prion assay in several hours to a few days, compared to months with the previous technique in animals. For more information please see their publication: Rapid End-Point Quantitation of Prion Seeding Activity with Sensitivity Comparable to Bioassays in PLoS Pathogen (6(12): e1001217. doi:10.1371/journal.ppat.1001217).

See BMG LABTECH’s Rocky Mountain Laboratory customer focus for more information on this new prion disease assay.

Wednesday, October 24, 2012

Fun Fact: This week is National Chemistry Week – Help Build Awareness of Chemistry

Twenty-five years ago the American Chemical Society (ACS)  started the National Chemistry Week (NCW) to help raise awareness about Chemistry for young people at the local level. This year the NCW has partnered with the Nanoscale Informal Science Education Network to spread the word about the growing field of Nanotechnology. C60 Buckyball croped
To help spread the word or to participate in events, please visit these NCW websites to find out more information:
- Like NCW on Facebook
- Follow NCW on Twitter
- Join NCW on the ACS network
- Find out what is going on near you
- Learn about some hands-on activities for kids

Tuesday, October 23, 2012

Did you know that the low volume LVis Plate, which is used for assays like DNA, RNA and protein concentration determination, is now compatible on the PHERAstar FS?

The LVis Plate from BMG LABTECH allows low volume (2 uL) samples to be measured in the absorbance detection mode. The LVis Plate is ideal for determining the concentration of DNA, RNA and proteins without the need to create a standard curve. Fully compatible with the SPECTROstar Nano and the Omega series of microplate readers, the LVis Plate is now fully integrated into the newest version of the PHERAstar FS software (v4.00). Included in the software are easy to use Wizards that walk users through Pathlength Calibration, Cleanliness Check (see Figure), and Blank Measurements for the LVis Plate. Taking low volume measurements in a microplate reader has never been easier.

Monday, October 22, 2012

FAQ: For absorbance assays, why should samples that are >3 O.D. be diluted and measured again?

For absorbance measurements, the optical density (O.D.) is a logarithmic measurement of the percent transmission (%T) and it can be represented by the equation, A = log10 100 / %T. Here is a graphical scale that represents this:

 
That means a sample with:
  • 1 O.D. allows 10% of light to be transmitted through the sample
  • 2 O.D. allows 1% of light to be transmitted through the sample
  • 3 O.D. allows 0.1% of light to be transmitted through the sample
  • 4 O.D. allows 0.01% of light to be transmitted through the sample


In other words, if a sample has an O.D. greater than 3, this means that only 1 photon of light out of 1,000 will be measured by the detector. Even with the most sophisticated instruments, this small amount of light is very hard to accurately detect above the background noise. Therefore, measurements above 3 O.D. will have greater error and will in turn be less accurate than measurements taken at a lower O.D. Thus it is always recommended to dilute samples that are >3.0 O.D. and then to factor in the dilution factor to the final measurement. This is also shown in the following graph, note how the measurement is no longer linear at high concentrations, which correspond to higher O.D. values.
 
 

Friday, October 19, 2012

Focus On Mitochondrial Toxicity – Simple Intracellular assay to test oxygen consumption

Mitochondria are one of the main places in cells were energy is produced via aerobic respiration. Mitochondrial toxicity can occur when drugs or diseases affect the mitochondria and prevent the proper production of energy via oxygen consumption. Therefore a method is needed to test mitochondrial function and toxicity, especially when trying to design potential new drugs.
One way to measure mitochondrial function is to measure oxygen consumption, but traditional methods are limited by low throughput, by using specialized equipment, and by measuring only extracellular oxygen.

The MitoXpress®-Xtra assay from Luxcel Biosciences overcomes these limitations by providing a phosphorescent based method that can measure intracellular oxygen consumption using the time-resolved detection found in many multimode microplate readers. When used with the Atmospheric Control Unit (ACU)  on the FLUOstar Omega, a real-time monitoring system can be created that measures the change in intracellular oxygen in a cell monolayer (see Application Note 223).

Find out more about the MitoXpress®-Xtra assay on our website: http://www.bmglabtech.com/technology/mitoxpress.cfm.

Thursday, October 18, 2012

Inside Applications: Assessing pancreatic trypsin activity


  • Trypsin activity as a measure for acute pancreatitis
  • Fluorimetric kinetic assay performed with the POLARstar Omega microplate reader
  • Only 1.5 µL of AMC-derivative trypsin substrate needed


Testing pancreatic trypsin activity in a POLARstar Omega microplate reader from BMG LABTECH is quick and accurate. It allows for a much higher throughput of the fluorescent 96-well microplate format samples and has reduced the expense of costly consumables required for testing pancreatic trypsin in a spectrophotometer.

Read the entire Application Note:
Assessing pancreatic trypsin activity using the POLARstar Omega Microplate Reader

Wednesday, October 17, 2012

Fun Fact: The 2012 Nobel Prizes Have Been Chosen



Congratulations to the new Nobel Laureates! Their contributions to our society are an example for us all.

Physics
Serge Haroche and David J. Wineland

Nobel prize medal Chemistry
Robert J. Lefkowitz and Brian Kobilka

Physiology/Medicine
Sir John B Gurdon and Shinya Yamanaka 

Literature
Mo Yan 

Peace
European Union (EU)

Economics
Alvin E. Roth and Lloyd s. Shapley

And in case you did not get enough, check out some even more fun facts about the Nobel Prizes: http://www.nobelprize.org/nobel_prizes/nobelprize_facts.html

Tuesday, October 16, 2012

Did you know that BMG LABTECH microplate readers can withstand strong gravitational forces and still function normally?


Studying the physiological effects of gravity fluctuations on plants, researchers at the University of Tuebingen in Germany use BMG LABTECH’s POLARstar  microplate reader in parabolic flights and in a Hyper-g centrifuge without any adverse effects on the instrument. 

During a parabolic flight, different stages of gravitation are obtained ranging from hypergravity (about 1.8 g) to microgravity (about 10-3 g). In one flight, 31 consecutive parabolas are flown. The gravitational effects of the parabolic flight can be seen in a simulated video (http://www.novespace.fr/fr,popup,parabole.html) and in a real video (http://www.youtube.com/watch?v=Lhu198E8z2U). Normal gravity is designated as 1g, and on the accent of the parabola the gravity goes to about 1.8 g (80% higher than normal). Then on the descent when the experiments are performed, the period of microgravity lasts for about 22 seconds.

Investigating the signaling pathway from the stimulus (no gravity) to the response (e.g. activation or deactivation of the expression of certain genes), the level of two important second messengers, Ca2+ and hydrogen peroxide, were monitored. Two cell lines derived from Arabidopsis thaliana expressing either Cameleon (a calcium sensor) or HyPer (an H2O2 sensor) were used, as well as a wild type. The calcium and hydrogen peroxide measurements were performed on a fixed POLARstar OPTIMA  in kinetic mode during the whole time of the parabolic flight.

Results show that the calcium levels increased with microgravity and decreased afterwards, signifying an effect on calcium flux due to changes in gravity. Some of the results can be found online here: http://www.bmglabtech.com/customer-focus/parabolic-flights.cfm. Follow up experiments were recently performed at higher gravity levels with the robust POLARstar microplate reader. Using the Hyper-g centrifuge at ZARM (The Center of Applied Space Technology and Microgravity) in Bremen Germany, the entire unit was centrifuged up to 10g, further demonstrating the instrumentation robustness.
POLARstar OPTIMA outfitted for the no gravity flight