Muse® Assays for Cell Health Analysis

In addition to using cell health assessment for elucidating disease mechanisms and therapeutic discovery, monitoring key indicators of cell health and performance helps establish uniform standards of cellular performance across long-term research studies. Knowing the performance profile of your cells prior to running your bioassay can mean the difference between valid assay results and wasted reagents, lost time and discarded data. Get truly quantitative, accurate data on the health of your cells. Make the Muse® Cell Analyzer and Muse® Cell Health Assays your everyday partners in cell health assessment.

Featured Kits

Muse® Count and Viability Kit (Cat. No MCH100102)

This simple, linear assay rapidly provides cell concentration and viability information. The assay uses a premixed reagent composed of two DNA-intercalating fluorescent dyes. One of the dyes is membrane-permeant and stains all cells with a nucleus. The second dye only stains dying or dead cells, whose membranes have been compromised. This combination allows for the discrimination of nucleated cells from those without a nucleus or debris, and live cells from dead or dying ones, resulting in both accurate cell concentration and viability data. Stained samples are then analyzed on the Muse® Cell Analyzer. The use of dual fluorescent probes that clearly identify all nucleated cells, live and dead, allows for greater sensitivity and accuracy compared to colorimetric methods.

The following data outputs are shown:

  • Viable Cell Count (cells/mL)
  • Percentage of Viable Cells
  • Total Cell Count (cells/mL)

Data output for the Muse® Count and Viability Assay. Healthy Jurkat cells were mixed with heat-killed Jurkat cells and stained with Muse® Count & Viability Reagent, and then analyzed on the Muse® Cell Analyzer. Data output include summary data (not shown) and optional dot plots (shown here). Reported statistics include viable cells/mL, % viability, and the total cells/mL. The left hand dotplot shows viability vs cell size; the right hand plot shows viability vs nucleated cells.

Anticipated staining pattern of the four cell subpopulations analyzed using the Muse® Count and Viability Assay.

ALSO AVAILABLE: Kits for analysis of Oxidative Stress, Nitric Oxide and Ki67 Proliferation. Please see www.emdmillipore.com/muse for further information.

Muse® Cell Cycle Assay (Cat. No MCH100106)

This assay allows for the facile, rapid, and quantitative measurements of percentage of cells in the G0/G1, S, and G2/M phases of cell cycle on the Muse® Cell Analyzer. Unlike traditional cell cycle analysis, which has tradition-ally required complicated instrumentation and training, the Muse® system enables users to easily determine cell cycle distribution on their benchtops. The Muse® Cell Cycle Assay uses the nuclear DNA stain propidium iodide (PI) to discriminate cells at different stages of the cell cycle, which differ in DNA content.

Fast, Easy Protocol

Simply fix in ethanol and incubate with Muse® Cell Cycle Reagent for 30 minutes. After the samples are analyzed on the Muse® instrument, the percentages of populations are automatically displayed, along with a histogram with three markers to demarcate the G0/G1, S, and G2/M cell cycle phases.

Cell populations identified:

  • Cells in the G0/G1 phase
  • Cells in the S phase
  • Cells in the G2/M phase

Data output for the Muse® Cell Cycle Assay. Jurkat cells were stained with the Muse® Cell Cycle Reagent and analyzed on the Muse® Cell Analyzer. Data output include summary data (not shown here), and a dotplot and histogram showing the cell cycle distribution.

Analyzing the impact of cell cycle-disrupting compounds with the Muse® Cell Analyzer. The Muse® Cell Cycle Assay can be used for a variety of cellular treatment conditions and to study the impact of cell cycle disrupting compounds. Nocodazole, a microtubule disrupter, leads to cell cycle arrest in G2/M phase; etoposide, a known anti-cancer compound, also causes G2/M arrest.

Muse® Autophagy LC3-Antibody Based Kit (Cat. No. MCH200109)

This kit enables quantitative analysis of autophagy using an anti-LC3 mouse* monoclonal antibody conjugated to Alexa Fluor®555, used to measure and track the levels of cytosolic and autophagosome-associated LC3 using flow cytometry. Also included is a selective permeabilization buffer, which discriminates cytosolic LC3 from autophagic LC3 by extracting the soluble cytosolic proteins while protecting autophagosome-associated LC3, thereby allowing its fluorescence to be measured by flow cytometry or imaging. Since autophagy is a constitutive cellular degradation process, the use of an autophagy detection reagent prevents the lysosomal degradation of LC3, allowing for quantification of its fluorescence.

* The antibody specificity/species cross reactivity is for human, but is also validated for mouse, rat, and hamster via western blot.

Data generated using the Muse® Cell Analyzer along with the corresponding Muse® software module provide statistical values measuring:

  • Mean Autophagy Value (for both control and test samples)
  • Autophagy Induction Ratio (test sample fluorescence relative to control)

HeLa cells were either starved for 4 hours to induce autophagy or kept under fed conditions, and samples were prepared using the reagents in the Muse® Autophagy LC-3 Antibody Based Kit. The fgure above shows the histogram plot comparing the control versus the target sample. Results are also displayed on the Muse® instrument in a summary page format (not shown). Here, the mean autophagy intensity for each sample is determined and the autophagy induction ratio is then calculated based on the ratio between the target sample fluorescence versus the control sample. In this cell population, there is a 6.4-fold change between the control sample (e.g. no autophagy in blue) when compared to the starved sample (e.g. induced autophagy in red), indicating the presence of autophagy.

Muse® RFP-LC3 Reporter Autophagy Assay (Cat. No. MCH200110)

Quantitate autophagy in single cells by tracking the autophagy protein, LC3, using this rapid, simple assay. This kit includes an mRFP-LC3 reporter cell line (stably transfected U20S human osteosarcoma cells that constitutively produce high levels of LC3) for comparative studies. Also included is a selective permeabilization buffer, which discriminates cytosolic LC3 from autophagic LC3 by extracting the soluble cytosolic proteins while protecting autophagosome-associated LC3. A monomeric RFP is used as a reporter to facilitate the translocation of the fusion protein.

Data generated using the Muse® Cell Analyzer along with the corresponding Muse® software module provide statistical values measuring:

  • Mean Autophagy Value (for both control and test samples)
  • Autophagy Induction Ratio (test sample fluorescence relative to control)
  • Percentage of cells with increased autophagy (test sample versus control)

Data output for Muse® RFP-LC3 Reporter Autophagy Assay. The RFP-LC3 reporter cell line was either starved for 4 hours to induce autophagy or kept under fed conditions, and then treated with autophagy reagents A and B (during sample preparation steps). Here, the mean autophagy intensity for each sample is determined and the autophagy induction ratio is then calculated based on the ratio between the target sample fluorescence versus the control sample. In this cell population, there is a 3.5-fold change between the control sample (e.g. no autophagy in gray) when compared to the starved sample (e.g. induced autophagy in red), indicating the presence of autophagy. Moreover, the percentage of increased autophagy is calculated between control and test samples, where it is calculated at 85.4% (not shown).

     
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