Millipore's Benchtop Flow Cytometer

September 02, 2016

Oregon Health and Science University
Biomedical Engineering
Graduate Research Assistant

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Company:

Millipore

Product Name:

Guava® easyCyte 12 Flow Cytometer

Catalog Number:

0500-5012

Millipore’s Guava® easyCyte 12 flow cytometer offers the performance of a traditional flow cytometer in a compact, benchtop-sized package. Our group has used the Guava extensively for well over a year to detect cell surface markers, perform cell cycle analysis and nanoparticle uptake studies. Part of our research involved determining different subpopulations of breast cancer cells within a single cell line before and after treatment with various anti-cancer agents, some of which are being developed in our lab. We were also interested to see how these agents affect cell cycle. Finally, we routinely monitor the cellular uptake of our nanoparticle drug delivery platform.

After comparing various candidate flow cytometers that were within the same price range with similar features, we ultimately chose the Guava because of the availability of a UV laser as well as the sheathless focusing of samples using capillary fluidics. The inclusion of a UV laser allowed us to use a greater range of dyes and fluorophores. In addition, without the need to supply sheath fluid, there is significant cost saving in term of consumables and it allows for smaller sample volume and more diluted sample while retaining sample acquisition speed. Finally, a calibration module that uses calibration beads are also provided to ensure that lasers and detectors are within specification before each run, without having to perform tedious laser alignment.

Overall, this is a great instrument with excellent capabilities at a competitive price.

Experimental Design and Results Summary

Application

Cell surface marker detection, cell cycle

Starting Material

Human breast cancer cell lines

Protocol Overview

Cell surface markers detection: Breast cancer cell lines BT474 and MDAMB231 were trypsinized and 1X10^6 cells of each were aliquoted into 1.5 ml tubes. Cells were washed with FACS buffer and then resuspended in 100 µl of FACS buffer containing FITC labeled CD44 and APC labeled CD44 antibodies. Cells were incubated with the antibodies on ice with rocking for one hour, after which they were washed twice with FACS buffer and then resuspended in 500 µl of FACS buffer for analysis.Cell cycle: Breast cancer cell line HCC1954 was trypsinized and resuspended in RPMI1640 medium with 10% FBS at a concentration of 1X10^6 cells per ml. Hoechst 33342 was added to a final concentration of 10 µg/ml and the cells were incubated at 37 °C for 30 min. Cells were then pelleted and washed once with FACS buffer. 1X10^6 cells were resuspended in 500 µl of FACS buffer for analysis.

Tips

Avoid too high cell density and perform regular cleaning of the capillary to ensure clog free operation.

Results Summary

A. Cell surface markers detection: CD24 and CD44 labeling of human breast cancer cells shows that BT474 is luminal (CD24+/CD44-) while MDAMB231 is basal and stem cell like (CD24lo/CD44+), which agrees with previously published data.B. Cell cycle: HCC1954 labeled with Hoechst 33342 shows distinct G1 and G2 peaks, which is comparable to that of propidium iodide staining.

Features Summary

Sheathless system with UV, blue and red lasers

Additional Notes

None

Image Gallery

Summary

The Good

Triple laser system at a very competitive price. Performance comparable to other flow cytometers.

The Bad

Capillary system more easily clogged when running large, clumpy cells at high density when compared to shealth based systems.

The Bottom Line

The Guava easyCyte is an affordable and highly capable system. With its compact footprint, it is definitively worth considering for your lab if you frequently perform flow cytometry analysis.

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