Biophysical Society 63rd Annual Meeting | Program Guide

Exhibitor Presentations Rooms 301 and 303, Baltimore Convention Center

Room 301: Sunday, March 3 10:30 AM – 12:00 PM HORIBA Scientific Unique Fluorescence Molecular Fingerprinting in Action: What Can CCD Detection Do for You? Fluorescence is a standard tool for the study of changes on the molecu- lar level, but it is now also becoming an emerging technique for molecu- lar fingerprinting and spectral kinetics. The Duetta™ 2-in-1 fluorescence and absorbance spectrometer from HORIBA Scientific is a unique and powerful benchtop instrument that provides so much more than standard PMT-based scanning benchtop fluorometers. CCD detection technology, and incorporated absorbance measurements, provide more data, with more accuracy, and in less time. In this presentation, HORIBA Scientific will demonstrate two of many methods for which Duetta is uniquely equipped to measure fluorescent samples. First, Duetta can measure protein binding and FRET over the full emission range (250- 1100 nm), demonstrating the effects of both donor and acceptor spectra over time with true spectral kinetics. In addition, the method of measur- ing Absorbance-Transmittance Excitation Emission Matrices (A-TEEMs) gives information about the molecular fingerprint of a mixture for use in component analysis of mixtures. The use of the absorbance detector enables inner-filter effect correction, which can easily be overlooked using standard fluorometers. Full Spectral Kinetics and FRET Because Duetta uses a CCD detector for emission detection, kinetics over the entire emission spectrum (250-1100 nm) instead of only at one or two different emission wavelengths. We will demonstrate the binding of a small molecule, 1,8-anilinonaphthalene sulfonate (ANS), to bovine serum albumin protein (BSA) that shows both the decrease in donor emission (BSA) and the increase of the acceptor emission (ANS) as an example of FRET kinetics. The binding of ANS to hydrophobic pockets in BSA is a known phenomenon, but is typically only measured as a kinet- ics experiment at the ANS emission wavelength of 475 nm. Historically, concentration-dependent experiments where emission spectra are col- lected over a range of ANS or protein concentrations, or both, are used to show binding kinetics or FRET as well. Duetta easily measures both the donor BSA (tryptophan) emission as well as the acceptor ANS emis- sion during binding and shows that energy transfer occurs over the full spectral range. This is a unique capability for a benchtop fluorometer in

a method of measuring the full fluorescence contour plot of a sample at all excitation wavelengths and all emission wavelengths. The matrix is then corrected for effects of high concentration (inner-filter effect) using the absorbance spectrum. The resulting A-TEEM gives an accurate profile of all emitting species and in turn, gives more information about the content of the sample in question, thus making it a better data set for chemometric and quantitative analysis. Solutions of tryptophan and 2-aminopurine, a fluorescent derivative of adenine, are used to demonstrate 1.) Effects of high absorbance/concentration on the fluo- rescence profile; and 2.) The A-TEEM profile for detection of multiple components. Speaker Karen Gall, Applications Scientist, HORIBA Scientific 2:30 PM – 4:00 PM IonOptix High-Content, High-Throughput Calcium and Contractility Measurements in Intact Cardiomyocytes High-content excitation-contraction coupling measurements in cardio- myocytes have historically been a slow, labor-intensive process requiring significant user involvement. While challenging, this methodology has proven itself essential throughout countless publications in the study of cardiac physiology and disease. Throughput, however, has limited the scope of calcium and contractility measurements and restricted study sample size and the number of conditions that can be tested in a given investigation. To improve the speed of data acquisition and analysis without compromising data quality, several advancements needed to be made to both the instrument hardware and software. Through its collaboration with IonOptix, CytoCypher’s MultiCell system improves on the traditional instrument by introducing many innovative approaches, including a cutting-edge fast motorized microscope and automated pro- cesses to improve throughput while preserving data fidelity. The new MultiCell system provides high optical and temporal resolution calcium and cell shortening data as well as automatic, “single-click” analysis. New features focused on pipelining data acquisition have improved the reliability and reproducibility of data collection. The resulting methodol- ogy is orders of magnitude faster, permitting investigations with greater statistical power and higher confidence. In this presentation, we will demonstrate the CytoCypher MultiCell high-throughput system for calcium and contractility measurements on intact, isolated myocytes. We will also show our protocols for these experiments, along with the analysis and statistical treatment of the resulting data sets. With this novel instrument, we have consistently acquired and analyzed data from over 1,000 cardiomyocytes per day. Speaker Michiel Helmes, CEO, CytoCypher/CSO, IonOptix

the field of biological fluorescence. A-TEEM Molecular Fingerprinting

The use of fluorescence for molecular fingerprinting is a relatively new concept and just as exciting if not more so than spectral kinetics. In most applications, changes in fluorescence intensity, or wavelength, or both, correlate to changes in physical properties of a sample. A-TEEM is

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