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LumiCycle

Luminometry for Circadian Biology

LumiCycle performs high-throughput luminometry on self-luminous tissues, such as those from transgenic animals containing the luciferase gene.

LumiCycle 32 Color records from 32 samples in 35-mm Petri dishes using 4 photomultipliers. Colored filters can be placed on 2 of the photomultipliers, so that 16 samples can be recorded simultaneously in 2 different colors.

LumiCycle 96 records from 96 samples in 4 24-well plates using 8 photomultiplier tubes.

LumiCycle In Vivo will record from an intact animal in a standard mouse cage using 2 photomultiplier tubes.

The systems are equipped with photon-counting photomultiplier tubes, each selected for low dark counts and high sensitivity in the green portion of the spectrum at which luciferase emits light.

The LumiCycle 32 and 96 fit inside a standard incubator. An internal fan circulates the incubator air to maintain the proper temperature within the chamber. The temperature is therefore as stable as the incubator can make it. The turntable and photon counting are fully automated. System setup and operation is straightforward. Online help files are included with the software.

In addition to its high-precision photon counting hardware, LumiCycle has the most flexible and easy-to-use software for the collection and analysis of circadian rhythms in luminometry data.

  • Start and stop each counting channel asynchronously. You may, for example, start one experiment with 10 samples on one day, and start a second experiment with 22 samples the following day, stop the first experiment 5 days later and start yet a third set of dishes in their place.
  • Interrupt the data collection to perform experimental manipulations on the samples (such as adding a drug). Once the treatment is complete, LumiCycle will pick up exactly where it left off.
  • Flexible counting schedules, from once per hour, to once per minute.
  • View records during the data collection process. A single window shows data for all 32 or 96 channels at once. Double-click on one of the graphs to view the data in greater detail (top figure).
  • Analysis program (bottom figure) automatically compensates for baseline shifts and signal attenuation over the course of the trial. The dominant circadian period and phase can then be extracted (red curve). View periodograms and actograms, calculate phase shifts, extract periods with spectral or periodogram methods.
  • Superimpose multiple records for comparison and figure preparation.
  • Calculate rhythm parameters for the complete record, or for multiple subsets of the record. If, for example, a treatment is given halfway through a record that is expected to create a period or phase change, the two halves of the record can be analyzed separately and a phase shift calculated.
  • While data collection is in progress, you may repeatedly copy the growing data files onto another computer (via a floppy or the network) and analyze the data collected so far for period and phase. Data collection continues uninterrupted until the end of the experiment.
  • Extracted periods and phases can be exported to a text file for later manipulation in Excel or other statistics programs.
  • Raw or baseline-subtracted records can be exported to ClockLab for further analysis using periodograms, actograms, and activity profiles.

LumiCycle 32#

32-Channel Luminometer for Circadian Biology

LumiCycle performs high-throughput luminometry on self-luminous tissues, such as those from transgenic animals containing the luciferase gene. The system is equipped with 4 photon-counting photomultiplier tubes, each selected for low dark counts and high sensitivity in the green portion of the spectrum at which luciferase emits light. 32 tissue samples, each in a 35 mm Petri dish, can be counted at one time. 8 dishes share each photodetector using a turntable device that alternately brings each dish under the detector, one at a time. (Specifications.)

The LumiCycle apparatus fits inside a standard incubator. An internal fan circulates the incubator air to maintain the proper temperature within the chamber. The temperature is therefore as stable as the incubator can make it. The turntable and photon counting are fully automated. System setup and operation is straightforward. Online help files are included with the software.

Two-color option. With the 2-color adaptation, colored filters can be placed on 2 of the photomultipliers so that 16 samples can be recorded simultaneously in 2 different colors. To create the image on the right, fibroblasts were transfected with Bmal-Nluc and Per2-Fluc, and recorded with a 550 nm long-pass filter on one PMT and 500 nm short-pass filter on the other. Optical filters can be easily changed by the user, or they can be removed for recording in the standard, non-color mode.

In addition to its high-precision photon counting hardware, LumiCycle has the most flexible and easy-to-use software for the collection and analysis of circadian rhythms in luminometry data.

  • Start and stop each counting channel asynchronously. You may, for example, start one experiment with 10 samples on one day, and start a second experiment with 22 samples the following day, stop the first experiment 5 days later and start yet a third set of dishes in their place.
  • Interrupt the data collection to perform experimental manipulations on the samples (such as adding a drug). Once the treatment is complete, LumiCycle will pick up exactly where it left off.
  • Flexible counting schedules, from once per hour, to once per minute.
  • View records during the data collection process. A single window shows data for all 32 channels at once. Double-click on one of the graphs to view the data in greater detail (top figure).
  • Analysis program (bottom figure) automatically compensates for baseline shifts and signal attenuation over the course of the trial. The dominant circadian period and phase can then be extracted (red curve). View periodograms and actograms, calculate phase shifts, extract periods with spectral or periodogram methods.
  • Superimpose multiple records for comparison and figure preparation.
  • Calculate rhythm parameters for the complete record, or for multiple subsets of the record. If, for example, a treatment is given halfway through a record that is expected to create a period or phase change, the two halves of the record can be analyzed separately and a phase shift calculated.
  • While data collection is in progress, you may repeatedly copy the growing data files onto another computer (via a floppy or the network) and analyze the data collected so far for period and phase. Data collectioncontinues uninterrupted until the end of the experiment.
  • Extracted periods and phases can be exported to a text file for later manipulation in Excel or other statistics programs.
  • Raw or baseline-subtracted records can be exported to ClockLab for further analysis using periodograms, actograms, and activity profiles.

LumiCycle 96#

96-Channel Luminometer for Circadian Biology

Like LumiCycle 32, LumiCycle 96 performs high-throughput luminometry on self-luminous tissues, such as from PER-LUC mutants. LC96 records from 4 24-well plates. The system is equipped with 8 photomultiplier tubes, each selected for low dark counts and high sensitivity in the green portion of the spectrum at which luciferase emits light. A turntable and moving PMT mounting alternately brings each sample under its detector. (Specifications.)

LumiCycle 96 is identical in size to LumiCycle 32. The apparatus fits inside a standard incubator. An internal fan circulates the incubator air to maintain the proper temperature within the chamber. The temperature is therefore as stable as the incubator can make it. The turntable and photon counting are fully automated. System setup and operation is straightforward. Online help files are included with the software.

Comparison of 96-channel and 32-channel systems:

  • In LC96, 12 samples share each PMT (instead of 8), so that each sample is counted for 2/3 less time during each hour.
  • The PMTs in LC96 are “end-on” instead of “side-on”.
  • The PMTs in LC96 have a larger window, which increases the background signal (dark counts) by about 2-fold, but also increases the signal itself by a comparable amount.
  • The light-tight cabinets are identical in size.
  • LC96 uses two USB slots on the host computer (instead of one).
  • File formats and analysis software are identical.
  • Both systems have very low cross-talk between channels. In LC96 cross-talk is less than 1 part in 1000.

In addition to its high-precision photon counting hardware, LumiCycle has the most flexible and easy-to-use software for the collection and analysis of circadian rhythms in luminometry data. As with LC32, recording is continuous; recording on each 24-well plate can be started and stopped independently.

  • Start and stop each counting channel asynchronously. You may, for example, start one experiment with 10 samples on one day, and start a second experiment with 22 samples the following day, stop the first experiment 5 days later and start yet a third set of dishes in their place.
  • Interrupt the data collection to perform experimental manipulations on the samples (such as adding a drug). Once the treatment is complete, LumiCycle will pick up exactly where it left off.
  • Flexible counting schedules, from once per hour, to once per minute.
  • View records during the data collection process. A single window shows data for all 32 channels at once. Double-click on one of the graphs to view the data in greater detail (top figure).
  • Analysis program (bottom figure) automatically compensates for baseline shifts and signal attenuation over the course of the trial. The dominant circadian period and phase can then be extracted (red curve). View periodograms and actograms, calculate phase shifts, extract periods with spectral or periodogram methods.
  • Superimpose multiple records for comparison and figure preparation.
  • Calculate rhythm parameters for the complete record, or for multiple subsets of the record. If, for example, a treatment is given halfway through a record that is expected to create a period or phase change, the two halves of the record can be analyzed separately and a phase shift calculated.
  • While data collection is in progress, you may repeatedly copy the growing data files onto another computer (via a floppy or the network) and analyze the data collected so far for period and phase. Data collection continues uninterrupted until the end of the experiment.
  • Extracted periods and phases can be exported to a text file for later manipulation in Excel or other statistics programs.
  • Raw or baseline-subtracted records can be exported to ClockLab for further analysis using periodograms, actograms, and activity profiles.

LumiCycle In Vivo#

LumiCycle In Vivo is designed to record PER-LUC light emissions from awake behaving animals placed in a standard mouse cage and infused with luciferin analogs. (Specifications).

For over a decade, it has been possible to record PER-LUC light emission from isolated tissues and cells. As methods and signals improve, it is now possible to record light emission from intact, behaving animals, opening up a wide range of experiments on the interactions among the clocks in different organs operating in their natural environment. LumiCycle In Vivo is a simple, robust apparatus for recording luminescence from intact animals, while simultaneously recording activity, monitoring light, temperature, and humidity within the chamber, and controlling the lights in infinitely programmable skeleton schedules.

Luminescence data from a PER-LUC mouse fitted with a luciferin mini-pump (Courtesy Prof. Mary Harrington, Smith College).

  • Two PMTs for even sensitivity across the cage area.
  • Red-shifted PMT sensitivity to accommodate the absorption of green photons by the intervening tissues.
  • Activity input for recording through a running wheel or IR motion sensor.
  • Programmable LED lights in each chamber for creating skeleton light schedules.
  • Light levels, temperature and humidity recorded continuously for animal care and use monitoring.
  • High-capacity internal fan for 15 air changes per hour.
  • Up to 8 units can be run on one computer. Recording on each unit can be started and stopped independently, and each can be given its own light schedule.
  • Data can be viewed and analyzed in the LumiCycle Analysis or ClockLab Analysis programs.

The LumiCycle In Vivo front panel, showing ongoing collection of counts for 4 animals (2 PMTs), together with temperature, humidity, light levels (Lux) and the current setting for the LED illuminator. The graph below is the second-by-second photon count for the currently selected channel (4).