Fluorescence/luminescence plate imager using a high sensitivity two-dimensional sensor (camera)

Measurement under uniform conditions with no time lag by simultaneous addition and reading in all 96 or 384 wells. FDSS/μCELL is a laboratory screening system that compactly integrates technologies developed in drug discovery screening, enabling a purpose-built system that is simple to use. Simultaneous measurement and analysis of the kinetics of a sample’s fluorescence or luminescence intensity in all wells at the time of compound addition are made possible by the high sensitivity two-dimensional sensor (camera) and dispenser head (96 tip type/384 tip type). Screening various compounds at high...

Applications 1. GPCR GPCRs (G protein-coupled receptors) play a major role in cell signaling, and many GPCR-targeted medical drugs have been developed. FDSS/μCELL is capable of detecting messengers, such as Ca2+ and cAMP, which are major contributors to the GPCR signaling system by using fluorescence and luminescence probes. FDSS/μCELL allows simultaneous dispensing and kinetic measurement of compounds in whole microplate wells, thus realizing high throughput screening. • Ca2+ measurement: Fluo-4, Fluo-8, Cal-520, Aequorin • cAMP measurement: Glo-Sensor 2. Ion channel Ion channel, a class of...

System components Combinations of components support wide range of applications. * Computer table is not included. Heater unit Highly sensitive two-dimensional sensor (camera) A high sensitivity/high speed camera with a wide sensitivity range from fluorescence to luminescence. Performs various assays with high throughput as a fluorescence/luminescence plate imager. Because all wells of the microplate are read simultaneously, there is no time lag in the fluorescent indicator or in measurement between wells after substrate addition. To measure rapid fluorescence kinetics, data can be captured at...

Fluorescence optical unit An optical system for fluorescence measurement that is integrated with a unique illuminator glass wave excitation optical system. It is used in combination with an LED excitation light source unit. It provides high S/N fluorescence detection that is maintenance-free with a long life. A complete line of excitation light source units can be easily replaced according to the purpose. Dispensing unit (96/384 tip type) Fluorescence optical unit M11031-02 A dispenser head that can dispense compounds simultaneously into all wells of 96/384 microplates. Since all wells are dispensed...

Measurement flow Provides flexible assay design and simple assay workflow Plate setting Simple-to-use manual settings Data acquisition Automatic setting for each company’s robot Protocol setting Call the assay protocol and set the number of measurements, measurement interval (measurement time), dispensing and washing conditions in the Kinetic Protocol mode. Operations from measurement to data output can be automated. : Pre-measurement processes (PreIncubation etc.) : Measurement ( : Addition, Dispenser Head operation) : Post-measurement processes (Tip/head exchange etc.) Number of images acquired...

Data analysis Various data processing and analysis are possible from the results of measurement Spatial correction between wells (spatial uniformity) Negative control correction Positive control correction Baseline subtraction correction (subtract bias) IC/EC graph calculation from multiple series (4 or 5 parameters may be selected) IC/EC graph calculation using Max, Min, Average and Max-Min in up to three time ranges in the same series Slope calculation to maximum range of 8 Max, Min, Max-Min and Ratio calculation to maximum range of 8 Analysis of calcium transient waveform of iPS cardiomyocyte...

Examples of measurement and analysis in typical applications 1. GPCR GPCR screening can be performed by intracellular Ca2+ assay, cAMP assay and β-arrestin assay. Intracellular Ca2+ assay Evaluation of ATP dose response using Fluo-8 AM-stained CHO Cell: CHO cell Dye: Fluo-8 AM (AAT Bioquest) Compound: ATP final 100 μM – 1 nM Evaluation of ATP dose response using CHO cells: 384 format Cell: CHO 4000 cells/well Dye: Calcium Kit iCellux (Dojindo Laboratories) Compound: ATP final 10 μM – 10 pM Aequorin assay Intracellular Ca2+ assay by luminescence using an aequorin-expressing cell line • Cell: Aeg-CHO...

2. Ion channel Ion channel screening can be performed using intracellular ion fluorescence indicators. Ca2+ channel assay Measurement of Ca2+ oscillation using primary neurons • Cell: Primary neuron (Rat embryonic day 17, cerebral cortex) (Rat Cortex Neuronal Cell, Wako Pure Chemical Industries) • Dye: Cal-520 AM • FDSS: FDSS/μCELL ImagEM 1×1binning exposure time 31 ms, Interval 31 ms, +37 ºC Evaluation of Ca2+ oscillation using primary neurons Cl- channel assay Cl - channel assay using YFP • Cell: CHO cell • Dye: Premo Haliden Sensor 10 min after compound addition 17031013 G17-H20 Ratio 0.-8-1.5...

3. Luminescence Luminescence screening can be performed using luminescent probes such as luciferase or aequorin. Luciferase assay Cell number evaluation using luciferase luminescence Cell number evaluation by Cell Titer-Glo KIT Cell number (cell number) It has high linearity and high sensitivity that can detect luminescence even with a small number of cells. cAMP assay cAMP evaluation using GloSensor • Cell: CHO cell • Kit: GloSensor Aequorin assay Ca2+ assay using aequorin-expressing cells h-CTZ Cell: CHO-K1 stably expressing apoaequorin with a mitochondrial targeting signal Substrate: h-coelenterazine...

4. BRET/FRET Screening of protein-protein interaction can be performed using fluorescence/luminescence energy transfer. BRET assay Control protein evaluation using NanoBRET NanoBRET control protein calibration panel • A control protein in which a HaloTag NanoBRET ligand is bound to a NanoLuc-HaloTag fusion protein • Five types of controls with different ligand binding rates By drawing the calibration curve, it is possible to check how much the coupling rate can be detected NanoBRETTM Control protein panel — 5 vials representing the following amounts of fractional occupancy: a. NanoBRETTM Control...