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Performance data shows that the Bigfoot Cell Sorter resolves fluorescent signals accurately and precisely. Application data demonstrates the level of gating resolution and sort purity needed for immunophenotyping analysis and immune cell isolation, even for rare populations. We also demonstrate the Bigfoot Cell Sorter’s ability to sort single cells into 96 and 384 well plates with high precision and accuracy.
For additional application data, download the application notes listed in this section and on the Resources page.
The Invitrogen Bigfoot Cell Sorter can resolve high dimensional data by unmixing the spectral signatures of overlapping dyes. This allows greater panel expansion and consequently the amount of information that can be gathered from each sample. We have demonstrated that this 28-color panel can be used to identify up to 20 different populations from one sample. From those populations, the Bigfoot instrument can sort six ways, simultaneously, with high efficiency and purity, including more rare subsets.
The Bigfoot Cell Sorter demonstrates exceptional capabilities in high-throughput plate sorting, achieving unprecedented sort speeds, precise deposition accuracy, and single-cell recovery down to small volumes. Innovative features such as 4-way sorting into 96-well and 384-well plates, built-in calibration, media detection imaging, and robust hardware set it apart from other instruments. The data presented here establish the Bigfoot Cell Sorter as a game-changing solution for researchers seeking consistent and efficient single-cell deposition into multi-well plates, enhancing workflows and paving the way for transformative discoveries.
Table 1. Elapsed sort time to complete a full 96-well plate using single-droplet four-way sorting.
96-well plate number |
Four-way elapsed sort time (seconds) |
---|---|
Plate 1 |
8.06 |
Plate 2 |
8.58 |
Plate 3 |
7.18 |
Plate 4 |
6.96 |
Plate 5 |
7.46 |
Average |
7.65 |
Table 2. Elapsed sort time to complete a 384-well plate using single-droplet four-way sorting.
384-well plate number |
Four-way elapsed sort time (seconds) |
Plate 1 |
16.10 |
Plate 2 |
18.08 |
Plate 3 |
17.15 |
Plate 4 |
17.35 |
Plate 5 |
18.23 |
Average |
17.38 |
Spherotech™ 8-peak beads run in key channels on the Bigfoot Cell Sorter show narrow, well-defined peaks.
Although polarization is a property of laser light that is often ignored in flow cytometry, it can be useful in differentiating certain cell types. For example, eosinophil granulocytes in human blood show relatively higher levels of depolarized laser light in a side scatter (SSC) detector due to their birefringent properties. Analysis of polarization has also been found useful in malaria diagnosis and marine phytoplankton studies.
All models of the Bigfoot Cell Sorter include additional FSC (forward scatter) and SSC (side scatter) detectors for depolarized light detection. The figure shows the use of the SSC depolarized detector to identify eosinophils in lysed human whole blood. This differentiation does not require staining, leaving cells undisturbed and saving money on fluorophores. By adding a stain for CD16, other cell populations can be identified as well.
Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
For Research Use Only. Not for use in diagnostic procedures.