Main ContentNovoCyte 3000 Flow Cytometer
The flow cytometer examines visible light scatter and fluorescence parameters for each cell. Forward scatter indicates the relative size of the cell, while side scatter indicates its internal complexity. Flow cytometry can measure the number of cells, the percentage of live cells, and the presence of intra- and extra-cellular markers. A few common applications are cell cycle analysis, apoptosis assays, and phenotyping of mixed cell populations.
When planning a flow cytometry experiment and choosing fluorochromes, it is important to know the fluorochromes' specific excitation and emission wavelengths, as well as the lasers, filters, and detectors available on the instrument. Below is the information for the NovoCyte 3000.
| NovoCyte 3000 Configuration and Fluorochromes |
| Excitation Laser | Channel | Bandpass Filter | Ideal dyes | Examples of Fluorochromes |
Violet 405 nm | V445 | 445/45 | Pacific Blue | DAPI, BV421, eFluor 450 |
| V530 | 530/30 | AmCyan | BV510, cFluorV547, AF430, eFluor506 |
| V572 | 572/28 | Pacific Orange | QDot565*, BV570 |
| V615 | 615/20 | Qdot 605 | BV605, eFluor 605NC |
| V675 | 675/30 | Qdot 655 | BV650, eFluor 700 |
| V780 | 780/60 | Qdot 800 | BV785, Qdot 800* |
Blue 488 nm | B530 | 530/30 | FITC | FITC, AF488, CFSE, GFP, YFP, BB515 |
| B572 | 572/28 | PE | PE, Cy3, eFluor 570, SYTOX Orange |
| B615 | 615/20 | PE-Texas Red | PI, PE-Texas Red, RFP |
| B675 | 675/30 | PerCP | BB660*, PE-Cy5, PI, 7AAD, PerCP-Cy5.5* PE-AF647 |
| B780 | 780/60 | PE-Cy7 | PE-Cy7, PE-AF750 |
Red 640 nm | R675 | 675/30 | APC | APC, AF647, AF660, eFluor660, DRAQ 5 |
| R780 | 780/60 | APC-Cy7 | APC-Cy7, APC-AF750, APC-H7, APC-eFluor780 |
It is important to remember that many fluorescent dyes are excited by more than one laser and fluoresce in multiple channels. It is very helpful to use a spectral viewer when planning an experiment.