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Agilent Technologies, Inc.

NovoCyte® Penteon™ Benchtop Flow Cytometer

Five Lasers, 30 Colors Plus Scatter Parameters

The new NovoCyte Penteon flow cytometer builds on its successful predecessor, the NovoCyte, and provides an expanded set of capabilities that accommodate today’s high-end and increasingly sophisticated multi-color flow cytometry assays. You now have the flexibility to choose from up to 30 fluorescence channels utilizing up to 5 lasers with up to 30 independent detectors.

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NovoCyte Penteon Flow Cytometer Features

Built upon the tremendous success of the NovoCyte® line of flow cytometers, Agilent now introduces the most advanced benchtop flow cytometer on the market today, the NovoCyte Penteon™.

  • Expanded flexibility with 30-fluorescence channel options using 5 lasers
  • High sensitivity and resolution
  • Intuitive and powerful software for data acquisition, analysis, and reporting
  • Smart-design functionalities and walk-away operation to simplify your workflow
  • Automation-ready capability for high throughput needs
  • Wide, 7-log dynamic range eliminates the need for routine detector adjustments

The NovoSampler Q can be integrated into different laboratory automation platforms and efficiently processes both FACS tubes (using a 40-tube rack) and 24-, 48-, 96-, and 384-well plates. The intuitive and industry leading NovoExpress software has been further advanced, providing an exceptional user experience in data acquisition, analysis and reporting.

The Ultimate Photodetector

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Silicon photomultipliers (SiPM) are solid-state, silicon-substrate-based, photon-level-sensitive semiconductor devices, with a 7.2 log dynamic range. Consisting of a compact array of avalanche photodiodes operating in unison, the SiPM is a compact detector with photon counting capability. The innovative optics designed into the NovoCyte Penteon incorporates 30 independent SiPM, which collect and process signals for each of its fluorescence channels.

Excellent scatter resolution to detect small particles

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NovoCyte Penteon scatter detection optics and signal processing electronics have been optimized to resolve particles down to 0.1μm in size. With such excellent resolution, platelets, bacteria, and various submicron particles can be readily identified and analyzed.

High reproducibility and stability

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The NovoCyte Penteon and NovoCyte Quanteon's fluidic system is designed to deliver high performance. When compared to other flow cytometers, the fluidic consistency and stability of the NovoCyte Penteon and NovoCyte Quanteon is unmatched. Other instruments utilizing peristaltic pumps are often subject to fluidic pulsation, causing inconsistency and inaccuracy in absolute cell counts.

Applications

Apoptosis Assay

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Apoptosis, or programmed cell death, is the process by which cells regulate how they die, activating specific pathways that cause the cell to shrink, condense, and eventually be cleared by phagocytosis. This is in contrast to necrotic cell death where cells die uncontrollably and fall apart, which can lead to detrimental effects such as the activation of an immune response. Therefore, apoptotic cells that die in a very orderly fashion limit disruption of nearby cells and tissue.
There are many ways to measure cell death and distinguish it from apoptosis or necrosis. These assays are easily quantified using the NovoCyte flow cytometer due to automatic compensation settings and a wide dynamic range of fluorescence detection which eliminates the need for any PMT voltage adjustments

Immunophenotyping

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Immune status is associated with disease state, treatment efficiency, and response to external stimuli such as vaccines. Immunophenotyping quickly identifies candidate cell types, sub-classes and functions. Monitoring the frequency of numerous immune cell population as well as the differentiation/activation status of specific cell subsets such as monocytes, NK cells, T and B cells is essential as they may influence the immunogenicity of a vaccine and its efficiency. The NovoCyte Flow Cytometer enables simultaneous quantification of multiple leukocytes for better understanding the immune status of patients and surveillance of the immune response to infectious disease.

Intracellular Protein Detection

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Detection and analysis of intracellular proteins allow for additional characterization of cell subpopulations and cellular processes. In order to analyze proteins not located on the cell surface, fixation and permeabilization of the cell is required. However, many phospho-specific antibodies are not compatible with many common detergent-based permeabilization methods used for intracellular staining. Special attention is needed when determining the proper fix/perm method for your phospho-specific antibody. The most common method uses 1.5% paraformaldehyde for fixation followed by 100% methanol for permeabilization. While this method works for many antibodies, please note it may not work for every phospho-specific antibody.
Additionally, identifying various cell populations in a heterogenous sample requires staining for phosphorylated proteins coupled with surface proteins. Special consideration must be given to the sensitivity of these epitopes to fixative, taking precaution to avoid damage to the epitope. Therefore, the sample may require staining for specific surface markers before fixation.

Cell Cycle Analysis

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Normal human somatic cells are diploids containing a constant amount of DNA. During cell cycle progression, DNA synthesis results in a doubling of total DNA content, followed by restoration of the normal DNA content after mitosis. Detailed cell cycle analysis can be performed to understand tumor cell differentiation, cell transformation and cell-compound interaction with the NovoCyte flow cytometer.
 
Figure: After treatment with 10 migrograms/M MG132 or 500 micrograms/M 5-FI for 16 hours/ A549 cells were analyzed for cell cycle distribution with the ACEA Novocyte flow cytometer. The the Novoexpress built-in cell cycle analysis module, the plot shows cells ni G0/G1 phase (green), S phase (yellow) and G2/M phase (blue). Compared to normal untreated cells, MG132 treated cells were arrested at G2/M phase, while 5-FU treated cells were arrested at G0/G1 phase.

Cell Proliferation

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Cell proliferation is an essential function and highly structured event that when unregulated, can cause disease. We can measure proliferation through absolute cell counts or with a dye, such as CFSE. When cells labeled with CFSE divide, the dye is partitioned equally between daughter cells and we can measure the loss of CFSE fluorescence over time as the dye is continuously diluted. The mean fluorescence intensity (MFI) of the dye was also plotted with cell concentration over time to show the inverse relationship between the two. This type of assay is often used to look at changes in T lymphocyte activation.
 
Figure: Measurement of proliferation in Jurkat T cells using CFSA. A) Jurkat T cells were labeled with CFSE and analyzed on the NovoCyte flow cytometer over time to measure cell division. Each peak is representative of an individual time point. B) Absolute cell counts are plotted alongside mean fluorescense intensity (MFI) of CFSE over time showing the dilution of signal as cells divide.

Testimonials

Joan Christiansen, H. Lundbeck A/S. (NovoCyte review from SelectScience)

“My best friend in the lab!!!”

“I use the NovoCyte® to screen compounds for induction of micronuclei in the TK6 cell line as part of our gene tox strategy. With the NovoSampler® and 96-well format, the assay is extremely robust and the results reproducible to a very high degree. The NovoCyte® is user friendly and creating new applications to support our immune tox strategy has been delightfully easy, even for a novice within the field of immunology.”

Matthias Schiemann, Technische Universität München

“Software is straightforward, and the software interface is easy to handle”

“This (NovoExpress®) software is straightforward, and the software interface is easy to handle. The implemented auto compensation and hierarchic tree structure is a highlight for effective organization of experimental data.”

Luz M. Londoño, Halozyme Therapeutics Inc. (review from SelectScience)

“Amazing instrument, it is the future of flow cytometry!”

“I have only wonderful things to say about the Novocyte®. It is incredibly easy to use, we get reproducible results, the software is the best one out there, the maintenance is super easy, and ACEA has excellent technical support! Can’t wait to upgrade to the 25-channel instrument!“

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Agilent Technologies, Inc.

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