IsoCyte™ and IsoCyte DL™
The IsoCyte™ is an automation-friendly benchtop laser scanning cytometer available in either a single or dual laser configuration. Laser lines can be chosen from 405, 440, 488, 532, and 640 nm excitation wavelengths. Four different emission wavelengths can be detected simultaneously with the highly-sensitive photomultiplier tubes. The platform is "format independent" and in 4-5 minutes can scan an entire plate from 6 to 1536-wells for highly miniaturized assays in addition to microscope slides.
The IsoCyte™ enables robust real-time assays for microarray, bead, cell and colony classification and enumeration in user-defined or commercially available formats. The instrument operates by rapidly scanning a laser across the bottom of a microplate or slide. The fluorescence emission from the laser-excited area is collected with a patented optical head and measured with photomultiplier tubes. The emission is mapped in 2-dimensions through time-based correlation allowing construction of a fluorescence image. The IsoCyte™ is not a microscope. It uses a telecentric scan lens with a large depth of focus, eliminating the requirement to focus or acquire multiple exposures to cover an entire field of view with 3-4 micron resolution. A light scatter mode allows generation of images from non-fluorescent objects (e.g. cell colonies) where addition of dyes is unacceptable.
IsoCyte™ is ideally suited for high throughput screening of many cell-based assays. The user specifies the image acquisition parameters to suit the necessary resolution and scanning speed for the assay. Optimized image analysis occurs during the scan to detect objects that produce signal above a user-specified background and that have passed through custom object filters. Size, shape, and brightness are among the filters used to define objects of interest (cells, colonies, or beads) while debris or other inappropriate objects are ignored. The IsoCyte's four emission channels allow multi-plexing by imaging in four colors in the intensity domain and two in the anisotropy domain.
The proprietary optics provide the unique ability to generate anisotropy measurements for microarrays, beads, and cells in addition to homogeneous solutions. Anisotropy is the degree of polarization of emitted light from a molecule after excitation with polarized light. Small, freely rotating molecules scramble their emission polarization while large or bound molecules retain the polarization of the excitation.
Imaging in the anisotropy domain allows the measurement of Fluorescence Resonance Energy Transfer (FRET) through depolarized emission. Fluorescent proteins have very high polarizations when excited directly by the laser but when FRET occurs this non-optical resonance depolarizes the emissions. In addition to measuring the emission ratio of the donor and acceptor, their anisotropy drop is also used to characterize fluorescence signal generated from FRET. This allows protein-protein interaction assays in living cells that can be measured on a cell by cell basis over time. FRET assays have been developed by IsoCyte™ users and we are continuing development of new assays making use of this technology with our partners and customers.
IsoCyte™SL and DL Laser Scanner Specifications
The SL system is a single laser system from the selection of laser lines offered. The DL system is a dual line laser system consisting of any two lines from the selection of laser lines offered.
- Laser lines and output powers
Laser line selectable through software - 405nm 50mW
- 440nm 40mW
- 488nm 25 or 50mW
- 532nm 50mW
- 640nm 40mW
- Detection Modes
User selectable through software - Fluorescence
- Anisotropy (polarized fluorescence)
- Enhanced laser scatter
- Detectors
Four independently adjustable photomultiplier tubes - 4 channel (color) intensity
- 2 channel (color) anisotropy
- Digital Acquisition System
40 MHz sampling rate
14 bit digitization
Dual digital signal processing - Sample Format
Standard Multiwell plate (clearbottom)
Microscope slides with adaptor - Dimensions
(W x D x H) 18.5 x 29 x 15 inches - Windows based PC environment
Dual-core processor
4GB DRAM
500GB Hard Drive - Integrated Data Analysis Package
- Integrated Automation Platform Ready
Caliper Twister II
Thermo Scientific CRS Polara
Velocity11 VWorks - 21 CFR Part 11 Compliant Software Option
The IsoCyte™ is covered under multiple US and international pending patents.