28 ANALYTICAL AND LABORATORY EQUIPMENT
Fig. 3. An example of the 64 bins of PMT data shown as spectral plots from both the 488 and 638 lasers.
the other fluorescent parameters allows for much higher sensitivity in assays. Tis is particularly useful when measuring the effect of pharmaceutical treatments on cells during in vitro assays.
Looking toward the future, the arrival of spectral flow cytometry with this new system will enable new methods of data analysis for the highly- dimensional datasets that are produced. Flow cytometry data analysis has traditionally been performed by sequential gating of cell populations based on two parameter histograms in a manual or semi-automated fashion. Te high-resolution collection in the SP6800 produces single cell fluorescence spectrographs. New data analysis
techniques will be able to perform spectral fingerprinting, where subpopulations of cells will be identified based on similarities of their total fluorescence spectra (Fig. 5). Tis allows for automated classification of cellular subpopulations that does not rely on sequential gating of populations.
Spectral flow cytometry is a powerful new technology that makes it easier to perform large multicolour studies, as well as perform new applications that were not possible on previous flow cytometers. Tese features make spectral flow cytometry an important tool for cellular analysis.
For more information ✔ at
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Fig. 4. Spectral un-mixing representation. Spectra from the sample are collected. Single stain controls are used as reference controls and are inputted into a spectral un-mixing algorithm, which then returns conventional dot plots for conventional analysis.
Fig. 5. Subpopulations of cells have different spectral waveforms. These waveforms are unique to the dyes selected in panel design and can be used to view sub-populations apart from the limited 2x2 analysis of conventional flow cytometry.
Marsha Griffin and Mike Zordan are with Sony Biotechnology Inc. in California, USA.
www.sonybiotechnology.com
Multimode microplate reader employs new optical concept
With the TriStar² S Berthold Technologies introduces monochromator technology in its basic multi-label microplate reader. The TriStar² S is capable of these reading technologies: UV/VIS absorbance; fluorescence; FRET; Time-resolved fluorescence; luminescence; BRET and BRET². TriStar2 S with multiple
measurement technologies employs the new optical concept All-4-One, for the first time enabling luminescence, fluorescence and absorbance measurements with highest respective sensitivity as known from dedicated instruments: less than 6 amol ATP per well and less than 0.3 fmol Fluorescein per well. Besides filters the instrument is equipped with a unique 3D double
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monochromator with high blocking efficiency and high f-number (increased transmission) for filterless absorbance measurements and fluorescence excitation. The instrument is characterised
by a new dual mode PMT detector with extremely low noise for both, fluorescence and luminescence measurements. While unsurpassed digital single
photon counting technology is applied for luminescence, in fluorescence measurements a new triggered analogue operation mode with virtually no electronic noise has been implemented. Furthermore optical filters can be used for luminescence measurements enabling BRET (eg, functional assays for GPCR research) and multi-coloured
luciferase reporter gene applications. The TriStar2 S can be fitted with
up to three reagent injectors – based on the proven and most accurate JET technology – and temperature control for the microplate compartment. Two of the injectors have their tips in reading position enabling the measurement of extremely fast reactions. The reagent compartment at the
front is easily accessible and may keep reagents cooled by the addition of crushed ice. Special reagent mounts provide a safe fixation of small reagent vials und ensure complete consumption of the reagents through inclination of the vial. Thus, the TriStar² S microplate
reader offers a multitude of possibilities and a wide range of applications.
Measurements of enzyme activities, phagocytosis, Calcium flux, cell viability, apoptosis, immunoassays, protein and DNA concentrations, protein-protein interactions as well as spectral analysis are only a few of the many uses of the new solution. The unit is operated via the proven
and intuitive ICE software. Single and multiple endpoint readings are possible as well as kinetic and scanning measurements. The data is displayed in numerical and graphical formats and may be exported into EXCEL or printed. For increased sample throughput
the TriStar² can be integrated into lab automation systems.
For more information, visit
www.berthold.com
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