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　 is an analysis software suitable for analyzing various time-series data and circadian rhythms, such as biological clocks, temporal changes in drug responses, and cell toxicity tests. Circadian rhythms, which were also recently recognized with the 2017 Nobel Prize in Physiology or Medicine, regulate physiological rhythms such as sleep and hormone secretion through the periodic expression of specifi c genes (clock genes). ATTO's Kronos series (Kronos HT, Kronos Dio) is a system that monitors the expression of these clock genes using the luminescence of reporter genes (luciferase) over time. KronoAnalyzer is software that utilizes time-series data*1 acquired by the Kronos series to perform tasks such as extracting peak/trough times and values, calculating the period, acrophase, amplitude*2, as well as basic statistical data such as mean, standard deviation, and coefficient of variation. It is not only capable of comparing sample data within a single fi le but also extracting data from up to three individually acquired fi les for overlaying graphs and conducting comparative analyses.*1. Time-series data: Data obtained by continuously acquiring the measurement values of the same sample.*2. The period, peak phase, and amplitude are calculated after detrending.Data processing settings● Noise fi ltering● Detrend * setting● Peak/Trough setting● Periodic calculation (cosiner method)● Background subtraction● Range of data processingAnalysis FlowGroup setting● Layout● Sample name● Number of specimens (n)● Group● Subgroup● Blank wellKronos HTExcelData File●　Kronos series measurement data (kht, dtl format)●　KronoAnalyzer analysis data (krd format)●　Excel data(with template)Analysis result● Peak/trough value● Period● Amplitude● Vertex phase (Acrophase)● Basic statistical data(Average value, standard deviation, coeffi cient of variation (CV value), etc.)Time-series analysis for biological clocks, temporal changes in drug responses, cell toxicity tests, etc.Intuitive and easy-to-use interface Perform comparative analysis between past and present data.Time-series Analysis Software* Processing to extract rhythm by eliminating the infl uence not derived from rhythm

Analysis softwareKronoAnalyzerThere are fi ve data processing items: "Noise fi ltering", "Peak/Trough", "Detrend", "Period", and "Multicolor Calculation". Checking the box will aff ect the graph and text data. Also, when you click a specifi c point on the graph, the time and value are displayed. You can easily check the analysis results while looking at the graph.検量線の作成Data analysis with intuitive and easy operationData analysis according to desired settingsKronoAnalyzercan adjust the data processing settings according to the target organisms, phenomena, and experimental systems, enabling analysis under optimal conditions. The data processing settings include "Smoothing," "Detrend," "Peak (Trough) Detection," "Data Processing Range," "Period Analysis," and "Background Subtraction."検量線の作成Click the graph you want to enlarge or overlayCheck data processing items to refl ect on the graphClick cell to sort displayIf you check the box of the data processing item, "Noise Filtering", "Detrend" and "Multicolor Calculation" will be refl ected in all graphs. If you check "Display all", the entire 96-well plate will be displayed in a list, and if you double-click an individual well, it will be enlarged in a separate window, making it easy to compare side by side (up to 5 wells per fi le). After selecting multiple wells, click the "Graph" button to overlay the selected wells. Wells can be selected in a wide range by dragging the mouse pointer, and wells can be freely added and deleted with the control/shift key and click.Deta processingIf you click the "Numerical display" button in the graph window, the analysis data corresponding to the displayed graph will be displayed. The displayed analysis data includes "Period/Acrophase/Amplitude", "Peak/Trough value", and Data processing itemDrag to select all at once ⇒ Display overlaid with the "Graph" buttonDouble click to enlargePeriodAcrophaseAmplitudePeak/Trough valueGroup mean value/Standard deviation/CV valueClick cell to sort display"Group mean value/Standard deviation/CV value". Click inside the cell of each item to sort in ascending/descending order. Analysis data can also be output in Excel/CSV format.Data processingSimple AnalysisKronoAnalyzer can process various time-series data in an optimal state and allows for simultaneous comparative analysis of numerous data from multiple fi les. Its simple analysis results, consisting of period, acrophase, amplitude, and peak/trough values, facilitate easy comparisons.By combining noise fi lters, you can remove spike noise using a median fi lter and eliminate fi ne noise using a moving average fi lter.Remove the trend of the time width specifi ed according to the experimental system. The peak/trough detection also specifi es the time width in the same way.Background noise values specifi c to the measurement device can be subtracted.You can set the analysis range to be separated (up to 2 divisions) by removing unstable data just after the start of measurement or by stopping the measurement temporarily due to drug administration, etc.AcrophaseFitting curveTroughAmplitudePeakPeriodEntire displayIndividual well displayMerge display

Group settings for easy comparison and analysis of various data解析ソフトウェア　KronoAnalyzerApplication DataKronoAnalyzer can open up to 3 fi les at the same time and compare and analyze the data measured in the past and the data measured by another device. By grouping and averaging samples according to measurement conditions, diff erences in peak/trough values, Acrophase, and Period become clear, enabling highly reproducible and highly accurate analysis. In addition, since the average value graph of each group can be displayed overlaid, it is easy to grasp the trend of the data visually.Casein kinase (CK) is said to control the circadian rhythm by phosphorylating proteins (such as PER/CRY) involved in the intracellular clock of the suprachiasmatic nucleus (SCN) in the hypothalamus. To investigate the eff ect of CK, Bmal1 promoter-luciferase (Bmal1-Luc) expressing MEF cells were treated with a CK1 δ inhibitor, and the expression of the reporter gene (luciferase luminescence) was monitored using Kronos Dio. BMAL1 is a transcription factor involved in the regulation of PER/CRY expression and is known to be indirectly infl uenced by CK. Analysis of the Kronos Dio measurement data using KronoAnalyzer revealed that as the concentration of the CK1 δ inhibitor increased, the period became longer, the acrophase was delayed, and the amplitude became smaller. Inhibition of CK1 δ suppresses the phosphorylation of PER, preventing its degradation by ubiquitin and stabilizing it. As a result, the period of PER is prolonged, and it is believed that, as a feedback from the transcription-translation of various clock genes that occur in coordination, the period of Bmal1 also increases.Click the graph you want to enlarge or overlayEff ects of CK1δ on circadian rhythms[Experimental Method]Bmal1-Luc expressing MEF cells were seeded in 35mm dishes and synchronized with dexamethasone treatment. After synchronization, they were stimulated with 0.3% DMSO or CK1 δ inhibitor (0.1-1.0 µM) containing medium for 2 hours. Subsequently, the medium was replaced with luciferin-containing medium (100 µM), and luminescence of luciferase was measured using Kronos Dio with a measurement interval of 20 minutes and a measurement time of 1 minute, with 5 replicates. The data obtained from the measurements were analyzed using KronoAnalyzer.Data can be added or deleted easily with check boxes.8 dish data of Kronos Dio96 well data of Kronos HT24 well data of Kronos HTEach well can be categorized according to the experimental conditions by giving the number of specimens (n) < subgroup < the group an arbitrary name. The plate layout is also set semi-automatically by simply clicking on the wells. Number of specimens (n) < subgroup < 1 to 192 samples for each group (two 96-well plates) can be selected. Also, if grouped, not only Kronos series fi les, but also Excel fi les (with templates) can be overlaid and comparatively analyzed within the same fi le.Apply data analysis processing with checkboxes.Even if many windows are opened, just click the desired fi le and it will be brought to the foreground.Supports various data fi lesGrouping can be set from 1 well = 1 groupIn addition to Period, Acrophase, Amplitude, and peak/trough values, the mean, standard deviation, and CV values for each group are automatically calculated. A lot of mixed data can be sorted with one click. Analysis results can also be output in Excel/CSV format.Groups and subgroups are displayed for each fi le.Graph overlay displayAnalysis DataPeriod became longerControlCK1 δ inhibitorPeriod became longer

Product specifi cationsLuminometer for 35 mm Dish with CO2 incubator functionAB-2550 Kronos DioProduct Name ・CodeKronoAnalyzer・3510155Avairable data fi les・Kronos HT/Kronos Dio/Kronos fi les (kht format/dtl format)・KronoAnalyzer fi le (krd format)・ Excel fi le (with template)*Up to 3 fi les can be displayed, compared, and analyzed at the same timeData display settingGraph display・Data display of all wells (up to 3 fi les)・Single/parallel data display of each well(Up to 5 wells per plate, maximum 3 fi les x 2 plates x 5 wells selected/parallel display)・Superimposed data display (maximum 192 groups (well) x 3 fi les)Text data display・Time series data, Peak/Trough, PeriodData processing setting･Noise filetering settings (Moving average/Median/Polynomial fitting)･Detrend processing setting (Subtraction of specified moving average)･Background value setting (Background subtraction derived from equipment)･Blank setting (Subtraction of blank wells)･Multicolor analysis settings (Supports multicolor emission samples)･Peak/Trough/Period calculation setting･Graph XY axis settings (X axis: 0 to 999 hours, Y axis: 0 to 2x109, full/auto scale, range specification)･Data processing range setting (up to 2 sections can be set)Group setting・Set the number of specimens (measurements) (n=1 to n=192)・Subgroup setting (Minimum subgroup: 1 well = 1 subgroup ~ Maximum subgroup: 192 wells = 1 subgroup・Group setting (minimum group: 1 well = 1 group ~ maximum group: 192 wells = 1 group)・Blank group setting (1 group)Grouping data analysis・Basic statistical analysis of each group: calculation of mean, standard deviation, coeffi cient of variation・Data analysis for each group: time-series data, peak/trough, cycle・Overlay display of graphs of selected groups and subgroupsAnalysis DataPeak/Trough･Peak/Trough value and time･Peak/Trough symbol display･Data sorting (ascending/descending order)Period calculation (cosiner method)･Calculation of Period /Amplitude / Acrophase (by cosiner method)･Data sorting (ascending/descending order)Data save format・Save analysis data (krd)・Save graph image (TIFF/JPEG/BMP/PNG)・Save data processing and analysis results (Excel/CSV)LanguageJapanese/English (supports language switching on the software)SystemWindows10/11 (64bit), Memory 8GB or more Recommended display resolution 1,920×1,080Multi-specimen live cell real-time luminescence measurement systemWSL-1565 Kronos HTRelated ProductsName WSL-1565 Kronos HT Detector PMT（cooling at 10℃）×2 unitsmeasuring containerClear-bottom 24-well plate (standard)Clear-bottom 96-well plate, 35mm dish (optional)No. of plates 2 plates（Independent detector for each plate）Cultivate conditionRT +5°C to 45°C (0.1°C steps), CO2 concentration control by CO2 gas mixing unit (settable range: 1.0 to 20%)Multicolor measurementColor separation measurement (models that support 3-color separation are also available)Size 650(W) x 520(D) x 340(H)mm, 40kgName AB-2550 Kronos Dio Detector PMT 350～670nmNo. of samples 8 samples（35 mm dish）Cultivate conditionTemperature RT -5°C to 45°C (1°C steps)CO2concentration control 5% (fi xed) humidifi cation sponge for humidifi cationMulticolorSeparate measurement is possible for up to 3 colorsSize 280(W)×400(D)×330(H)mm　・　16.0kgSpecifi cationsRelated products3-2-2 Moto-asakusa, Taito-ku, Tokyo 111-0041, JapanTEL +81-3-5827-4863FAX +81-3-5827-6647URL https://www.atto.co.jp/engATTO Corporation