Instruments
BD FACSAria-SORP
The FACSAria-SORP cell sorters are capable of 12 fluorescence parameters. The sorters
have 4 laser lines, including a 60 mW 350 nm UV line, 100 mW 488 nm, 100 mW 561 nm,
and 40 mW 640 nm lines. The sorters can be used to isolate cells in multiple formats,
including 4-way tube sorting and 96-well plate sorting via the Automated Cell Deposition
Unit.
BD FACSCanto II
The FACSCanto II cell analyzers are equipped with 3 lasers (30 mW 405 nm, 20 mW 488
nm, and 17 mW 640 nm) capable of 8 fluorescence parameters. The analyzer is available
to individual investigators. Appropriate dichroics are available for numerous routine
as well as specialized assays for each of the instruments.
Nexcelom Celigo microplate based imaging cytometer
The CeligoS imaging cytometer is designed for performing assays directly in multi-well plates (6-384 wells). There are many advantages to this approach, including lower demand for cells (allowing for the use of rare primary cells isolated from patient samples) and reagents as well as analyzing the cells in their native state without the need to enzymatically remove them from their growth substrate, as is required for traditional flow cytometry. It can image cells in Brightfield and 3 fluorescent channels: blue (377/50nm Ex 470/22nm Em), green (483/32 Ex 536/40 Em), and orange/red (531/40 Ex 629/53 Em). Using advanced software based image recognition, there are over 10 specific analysis modes for a wide variety of applications. The CeligoS is also a self-serve instrument after training is provided.
XRAD320
https://www.kimtron.com/
On-Chip Biotechnologies Microfluidic Cell Sorter
(3 laser 6 color)
The On-Chip is the world’s first in its class microfluidic cell sorter and was developed in Japan. USA MCI was one of the first centers in America to obtain this new technology. One benefit over traditional sorting is the ability to analyze extremely low starting sample volumes/number of cells. Additionally, the sorting is much gentler and can accommodate a wide range of particles from exosomes to cell spheres using two different size microfluidic chips. The sorter is aerosol free and placed in a BSL-2 cabinet, allowing for the safe isolation of cells infected with lenti virus or other pathogens. The 3 excitation lasers (405nm, 488nm & 561nm) are collinear, so fluorochromes excited by any laser can be detected in any of these channels: FL1(445/20 nm), FL2(543/22 nm), FL3(591.5/43 nm), FL4(607/36 nm), FL5(676/37 nm), and FL6(732/68 nm).
On-Chip Biotechnology SPiS
The SPiS companion instrument does single cell or sphere deposition into 96 or 384
well plates in a far more gentle and reliable manner than traditional sorters.
ThermoFisher Neon Electroporation System
Designed for primary cells and large size plasmids, very gentle. Available for use
free of charge with appointment.
Miltenyi Biotech GentleMACS tissue disassociation system
The industry standard for preparing single cell suspensions from most any tissue with the highest viability. Available for use free of charge with appointment.
The ZetaView® TWIN - NTA Nanoparticle Tracking - Video Microscope PMX-220
The 2-laser platform (488 & 640 nm) was installed in the core lab in May 2019. Nanoparticle
Tracking Analysis (NTA) captures the Brownian motion of each particle in the video.
Based on the different diffusion movements of large and small particles in the surrounding
liquid, the hydrodynamic diameter of the particles is determined. Pattern parameters,
such as intensity fluctuations, surface geometry, and shape of the particles as well
as particle concentration are documented at each recording and can be used to distinguish
sub-populations. In addition, the charge state of the particle surface (zeta potential)
can be measured via the movement of the particles in an applied electric field. Depending
on the type of sample and the measuring mode, the measuring range is between 15 nm
and 5 μm. User rates are $25/hour for both university and outside the university users.
The Agilent Seahorse XFe24 Analyzer
Installed in the MSB lab in November of 2019, the Seahorse measures the oxygen consumption
rate (OCR) and extracellular acidification rate (ECAR) of live cells in a 24-well
plate format. Seahorse XF technology measures the flux of oxygen and protons in the
transient microchambers created by specially designed tissue culture microplates in
real time. The software calculates the rates of oxygen consumption and extracellular
acidification from its measurements of oxygen and proton flux. Usage is currently
covered under S10 OD025089-01A1.
Kimtron IC-320
https://www.kimtron.com/
10x Genomics Chromium Controller
The newest addition to FCSRLs fleet of instrumentation was installed February 2020. The platform uses advanced microfluidics to perform single cell partitioning and barcoding in a matter of minutes. The controller combines a single cell with a single gel bead that is coated with a unique oligonucleotide barcode to form a single cell emulsion droplet, which is referred to as a “GEM” (Gel Bead-in-emulsion). Within each GEM the unique barcoded oligonucleotides are used as primers to generate cDNAs, which can be traced back to cell of origin. The barcoded amplified fragments from hundreds to thousands of cells are pooled to create short read sequencer compatible libraries. Sequencing of these libraries enables analysis of single cells for gene expression, cell surface proteins, immune clonotype, antigen specificity, and chromatin accessibility. There are currently no user fees associated with this system, but advanced scheduling is required. Please call (251) 341-3030 to set up an appointment.
Miltenyi Biotech GentleMACS tissue disassociation system
The industry standard for preparing single cell suspensions from most any tissue with the highest viability. Available for use free of charge with appointment.
Cryostat
A cryostat (Thermo Scientific) is available for sectioning frozen tissues. Frozen
sections can be examined by various histological and immunohistological methods.
Finesse 325 Microtome
Formalin-fixed paraffin embedded (FFPE) tissues can be sectioned using the Finesse
325 microtome (Thermo Scientific) located in the MSB facility. FFPE sections can
be examined using various histological and immunohistological approaches.