Circadian Integration Explained for clock pathways

Kronos HT is a multi-well plate compatible model
appeared in Kronos series which measures
changes in gene expression over time
with bioluminescence of luciferase while
culturing cells.

ATTO High throughput real-time luciferase assay system
WSL-1565 ATTO Kronos HT

High throughput real-time luciferase assay system Kronos HT
High throughput assay system
for real-time gene expression monitoring of living cells
WSL-1565 Kronos HT is the high throughput support model of the assay
system that measure the gene expression of living cells in real-time with the bioluminescence of
luciferase. Kronos Dio, which is a precedent model compatible with 35 mm dish, has been used
as a real-time luminescence measurement system with cell culturing function, but this time a new
lineup of model compatible with multiwell plate measurement is added.

With this new product Kronos HT, we succeeded the know-how cultivated in the precedent model
in order to measure the luminescence while culturing the cells, and further strengthened functions
and performance for high throughput measurement. Please consider adopting Kronos HT for
high-throughput measurement in biological clock analysis, change in drug response with time,
cytotoxicity test etc.
NEW
• Cell culture function with temperature contol, humidifying
and controlling CO2 concentration
• 24 and 96 well plate format
• Scan 2 plates with 2 detectors (PMTs)
• It does not shake or vibrate cells by scanning with
detector moving and cuture plates not moving.
Biological clock, organ clock and cell clock... Biological clock, organ clock and cell clock.
chronomedicine, research progresses in the world.
Time
Intensity
Reporter assay in real time with culturing cells
Reporter assay is used to measure the transcriptional activity of gene transcription
regulatory sequences such as promoters, for researches on drug response and gene
the method of measuring the amount of bioluminescence using bioluminescent
enzyme luciferase using a device called a luminometer is superior in terms of high
sensitivity and low background and is widely used.
Usually, transcription activity at a certain point (end point) is measured, but the
assay is also becoming used for tracking transcription activity over time such as
stimulation response, time course of transgene expression level, periodic variation
of biological clock gene. Kronos HT is a system to optimize for such assays with the
ability to monitor the bioluminescence over time as cells are cultured.
Time course of drug response
Long-term kinetics of activity inhibition
Monitoring of biological clock
• High sensitive and low noise detection by cooling of
detectors (PMTs)
• Compatible with multicolor luciferase assay
• Easy setting with software on Windows PC, and
displaying and saving measurement data in real time.
chronomedicine, research progresses in the world.
expression control, evaluation of gene transfer effciency, and so on. 

EzFluoroStain DNA（緑）
Light detecting with culturing cells
Sample stage (CO2
incubator)
Filter wheel
Cooled PMT
Multicolor assay
Maintain cell culture environment
The sample stage is a CO2 incubator with
CO2
control.
24/96 well plate compatible with adapter exchange
Although it is compatible with 24 well plates in the standard model,
it is also able to use 96 well plates by simply replacing it with an
adapter of an optional item.
High sensitive detection
Weak light can be detected by cooling
the detector (photomultiplier tube, PMT)
to 10°C to suppress thermal noise.
Since two plates are scanned simultaneously by two
detectors, more samples can be measured at shorter
intervals.
Cell-friendly scanning mechanism
Since it has a mechanism to move the detectors
without moving the sample stage, the cultured
cells are not shaken or vibrated.
Heating and
humidifying the gas
Controlling
CO2
gas concentration
CO2
gas
Exchanged to a 96 well plate adaptor (option)
Sample stage (setting two of 24 well plates)
It can be applied to multicolor luciferase assay with filter
automatic switching mechanism
and ATTO's original color separation system (patent).
Software
Setting measurement conditions
You can set it by simple operation. Setting
operation can be omitted by using the function
Display measurement data in real time
Since the data under measurement is displayed
in real time, you can check how the measured
value changes over time. Measurement data
taken out even during measurement.
Data processing
It has a smoothing function to remove
noise component from data and smooth
it, a multicolor calculation function to
calculate each color emission value
and a detrending function to extract
rhythm variation of biological clock.
Data export
Since data can be exported as a CSV format
spreadsheet software such as Microsoft
Excel or other analysis software. Also, you
can export the graph list of each data as an
Multicolor calculation
Smoothing
Detrending
Export as
Graph list display of all samples
Detailed graph list display
Main window
Setting window
Circadian rhythm of clock gene expression
Application
NF-κB response elemnt - TK promoter - SLG (green color emited luciferase) and TK promoter -
e in 96 well clear bottom plate
Measurement time: 5s x 2 color / well, Interval time: 30min
(A) Transcriptional activity of NF-κB response element for 48h
(B) Transcriptional activity of TK promoter for 48h
(C) Normalized by activity of control (TK promoter)
(D) Ratio to NF-κB activity without TNF-α
Transcriptional activity of clock gene, mPer2
mPer2
stable cell line in 24 well clear bottom plate
Measurement time: 5s / well, Interval time: 10min
Dual-color real-time luciferase assay of TNF-α induced NF-κB transcriptional activity
（
Measurement Photon counting method
Measurement time Integral time: 1 - 60 s/well, Measurement term: 1 h - 30 days
Plate format Clear bottom 24 well plate x 2
Clear bottom 96 well plate x 2 (Option), 35mm dish x 12 (Option)
Throughput
48 samples (24 well plate x 2)
192 samples (96 well plate x 2) with an optional item, 96well plate adaptor
12 samples (35mm dish x 12) with an optional item, 35mm dish adaptor
Incubation
temperature control Room temperature + 5°C to 45°C (0.1°C step) at room temperature 20 - 28°C
CO2 gas control Control to 5% by CO2 gas mixer
2
intermittently by humidifying unit, Maintaining >90%RH
Multi-color assay
rs
(There is also a model for 3 color separation. Request inquiries.)
PC requirements
OS: Windows 10 / 8.1 / 7, 64 / 32 bit, Memory: more than 4 GB, HDD: more than
20 GB free space, Interface: USB2.0 x 1 port
Dimensions, Weight
Main unit: 650 (W) x 520 (D) x 340 (H) mm, 40 kg
CO2 gas mixer: 160(W) x 300 (D) x 200 (H) mm, 4.8 kg
Humidifying unit: 170 (W) x 140 (D) x 115 (H) mm, 1.6kg
Power
Main unit: AC 100 - 240 V, 50 / 60 Hz, 400 W (max)
CO2 gas mixer: AC 100 - 240 V, 50 / 60 Hz, 100 W (max)
Humidifying unit: AC 100 - 240 V, 50 / 60 Hz, 24 W (max)
Code No. Model
3510140
WSL-1565 Kronos HT
Kronos HT main unit, Control software (for Windows PC), CO2
gas mixer,
Humidifying unit, 24 well plate adaptor
Code No. Model
3510147 96 well plate adaptor
3510148 35mm dish adaptor
Option
CO2 Main unit PC Humidifying unit gas mixer
(PC is not included in standard components)
Real-time luciferase assay system
35 mm dish format model
AB-2550ATTO Kronos Dio
Related product
Code No. Model
3510120 AB-2550 Kronos Dio
> Reporter assay in real time with culturing cells
> Cell culture function with temperature control, humidifying and
controlling CO2
gas concentration
> 35 mm dish format, 8 dishes
> Compatible with multicolor luciferase assay
> Easy setting with software on Windows PC, and displaying and saving
measurement data in real time.
Model AB-2550 Kronos Dio
Detector PMT (Photomultiplier tube)
Measurement Photon counting method
Throughput 8 samples (35 mm culture dish x 8)
Measurement
time 1 s - 60 min / dish
Incubator Temperature setting: 20°C (RT - 5°C) to 45°C (1°C step), CO2 concentration: 5%
Multi-color assay

requirements
OS: Windows 10 / 8.1 / 7, 64 / 32 bit, Memory: more than 4 GB, HDD: more than
20 GB free space, Interface: USB2.0 x 1 port
Dimensions,
Weight 280 (W) x 400 (D) x 330 (H) mm, 16 kg
Power AC 100 - 230 V, 50 / 60Hz, 150W (max)
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The EasyNAT® System

 PCR POCT Workstation based on three-stage magnetic conductivity extraction technology and patented Cross Priming Amplification Technology.

 The system is available in a 4, 8 or 16-module configuration.

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