The Guava EasyCyte™ System is an automated, easy-to-use, desktop single-cell analysis system that can perform a wide range of multi-color cellular and bead assays. The cytometer has two lasers: Blue (488nm) and Red (640nm) and can detect up to six fluorochromes other than the forward and side scatter lights. Sample uptake occurs though a micro-capillary and does not require sheath fluid or other supplementary fluids for operation.
Fluidics Specifications and lasers/colors are detailed in the tables below. Instructions for using the instrument are in the room, but basic training is required before using. Please contact Ms. Maria Esmerian for details.
Lasers and Parameters
|
| Blue Laser (488nm)
| Red Laser (640nm)
|
| Max Emission |
Green (525/30 nm) |
Yellow (583/26 nm) |
Red (690/50 nm) |
NIR (785/70 nm) |
Red2 (661/19 nm) |
NIR2 (785/70 nm) |
FSC/SSC | | | | | | | |
7-AAD | 647 | | | X
| | X | |
Alexa Fluor 488/FITC | 519 | X | | | | | |
Alexa Fluor 647 | 668 | | | | | X | |
APC | 660 | | | | | X | |
APC-CY7 | 774 | | | | | | X |
Calcien | 515 | X | | | | | |
CFSE | 517 | X | X | | | | |
DiOC6 | 501 | X | | | | | |
PE-CY5 | 670 | | | X | | | |
PE-CY5.5 | 692 | | | X | | | |
PE-CY7 | 774 | | | | X | | |
PerCP | 675 | | | X | | | |
PerCP-CY5.5 | 690 | | | X | | | |
PE-Texas Red | 613 | | X | X | | | |
PI | 617 | | X | X | | X | |
Qdot525 | 525 | X | | | | | |
Qdot655 | 655 | | | | | X | |
Qdot800 | 800 | | | | X | | X |
Rhodamine 110 | 520 | X | | | | |
|
Fluidics Specifications
Flow Cell Dimensions | Standard square capillary with ID of 100µm |
Sheath Fluid | None |
Sample Flow Rate | 7µL/min to 72µL/min |
Waste Generation | Typically <40mL in 8 hours of continuous use |
Sample Concentration | Final particle concentration of 10,000 to 500,000 particles/mL |
Sample Requirement | As few as 2,000 cells/test; typically 25,000-100,000 cells |
Cell Size | 1-60 µm |
Typical # of Cells/Protocol | 1,000-10,000 cells/test
|
Software
GuavaSoft™ 2.7 Software includes dedicated modules for cell- and bead-based assays. GuavaSoft Software automatically saves data files as FCS format compatible with standard flow cytometry analysis applications (FlowJo, DIVA). An offline data analysis workstation is available in the room 308.
Reservation
Reservations of the cytometer via the online booking system are possible. You can access the reservation sheet under “Facility Reservation/ Faculty of Medicine/ Millipore Guava EasyCyte8".
Reservations are subject to approval by the operator and users are advised to verify their reservations 24 hours prior to the experiment in case of an emergency cancellation.
Bookings should be cancelled as early as possible in case of a cancellation of an experiment.
Application
The Guava EasyCyte™ 8 Flow Cytometer can be used to carry out multiple experiments across different research areas. These include:
- Accurate cell count and population determination
- Transfection efficiency
- Cell death assays (Annexin V/PI)
- Cell cycle/DNA content analyses
- Fluorescent dye transfer techniques
- Functional assays (proliferation, ROS production, membrane potential changes, …)
Operating the Guava Machine
Download the protocols here
Billing Fees
The billing calculation used by the Flow Core is done by the following:
At the end of the month, the amount of time reserved vs. the amount used for each reservation is checked. The greater of the two is billed for. The total amount of time used each month is then listed by the user and sent to the department for bills at the end of the month
- User GuidelinesAll reservations must be canceled 48 hours before reserved time via AUB Facility Reservation.
- If you are more than 15 minutes late and have not notified us that you will be late, we reserve the right to cancel your experiment and bill you for the reserved time.
- For all analytical instruments (Guava or Diva/ Flow Jo offline station), it is acceptable to shift your time at no charge but make sure not to overlap another user's scheduled time. Don't forget to cancel your reservation before re-booking a new time to avoid a cancellation fee.
- For BD FACS Aria cell sorter, shifting of time will result in billing according to the scheduled time.
- Please do not make reservations for a colleague /lab mate. Only trained and approved personnel will be granted access to the facility.
If you have any questions or concerns, please don't hesitate to contact us directly to discuss them.
What to Bring with You
- Samples should be brought in 1.5ml microcentrifuge tubes. You have to cut off the cap of the tube before loading it in the tube holder, or use a twist off cap tube. The type of plastic does not matter.
- Micropipette and tips for homogenization of the sample before loading.
- Your compensation controls.
- A CD/DVD to record your data.
Performance Check
The easyCheck Procedure is performed once per month to ensure that the system is performing properly. easyCheck averages the results from three acquisitions of a guava easyCheck Bead sample to determine if the results are within the expected range.
Hours of Operation: 8:00am-5:00pm
Reservation Link
Custodian:
Dr. Samia Khoury and
Dr. Layal El Hajjar
Access Level: Advanced
Location: DTS, 3rd floor, room 308