BD FACSAria III is the latest of all the flow cytometers available. It is unique in its ability to allow multiparametric analysis of the single sample population. It is highly sensitive and accurate in quantitative assessment of fluorescently labelled cell population. Having the ability to rapidly analyse thousands of cells per second attributes to its high precision. The BD FACSAria III flow cytometer is a high speed fixed alignment benchtop cell sorter. With its fixed-optics design and digital electronics, the BD FACSAria III flow cytometer enables multicolor analysis of up to 18 fluorescent and two scatter options at a time. The BD FACSAria III system consists of three major components: a fluidics cart, a benchtop flow cytometer and a workstation. Nearly all cytometer functions are operated from within BD FACSDiva software.
Flow Cytometer
The benchtop flow cytometer contains all the major components of the three subsystems (fluidics, optics and electronics). It is relatively compact than most of the sorters with the same capabilities.
a) Fluidics System
When fluidic system is activated the sheath fluid from the pressurized sheath tank is forced into the cuvette flow cell where hydrodynamic focusing forces particles from the sample injection chamber through the cuvette in a single file stream. Within the cuvette flow cell, laser light is focused on the sample core stream. Fluorescent molecules are excited by the different laser wavelengths thereby detected by the optics and analyzed by the electronics. Particles are then either transported into waste reservoir via waste aspirator or sorted into collection device. The integrated nozzles are available in three sizes (70, 85 and 100 mm) to accommodate a variety of particle sizes, plus a closed-loop nozzle for use in cleaning and shutdown procedures.
b) Optics System
The excitation optics consists of lasers, fiber optics cables, beam-shaping prisms and an achromatic focusing lens. The BD FACSAria III uses air cooled and solid state lasers that don’t have special power and cooling requirements. The system supports six installed lasers, out of which only four may be used at once. Further the laser light is detected by the collection optics that are arranged in octagon and trigon shaped arrays. Maximum signal detection is achieved by transmitting light to a series of photomultiplier tubes (PMTs) and finally to the detectors.
| Laser | Wavelength (nm) | Comments |
| Red | 633 | Standard |
| Yellow-Green | 561 | Opetional |
| Blue | 488 | Standard |
| Blue - Violet | 455 | Opetional |
| Violet | 405 | Opetional |
| Near UV | 375 | Opetional |
c) Cytometer Electronics
The electronics consists of power panel, the main power button turns on the cytometer and the fluidics cart. Lasers require atleast 30 minutes to warm up. Emergency stop button is a safety feature which when pressed stops the movement of the loading port and deflection plate voltage is switched off. Data acquisition and analysis are controlled by BD FACSDiva software.
Applications
a) Cellular uptake studies- Used for the quantitation of various fluorescently labelled drugs/ molecules into the mammalian as well as plant cells. It allows us to perform multiparametric analysis in the same set of cells with higher accuracy and reproducibility as compared to other techniques.
b) Cancer Studies- Measures the number of cells in a sample, the percentage of live cells and dead cells can also be analyzed when treated with certain dyes such as propidium iodide (PI). The presence of tumor markers on the cell surface can also be detected when labelled with tumor specific fluorophore/ antibodies.
c) Detection of apoptosis with Annexin V- Annexin V conjugates provide quick and reliable detection for studying the externalization of phosphatidylserine, an indicator of intermediate stages of apoptosis. The difference in fluorescence intensity between apoptotic and non-apoptotic cells stained with our fluorescent annexin V, as measured by flow cytometry, is typically about 100-fold.
d) Cell Cycle analysis - Cellular DNA content can be analyzed following cell staining with 4',6'-diamidino-2-phenylindole (DAPI). This approach reveals distribution of cells in three major phases of the cycle (G1 vs S vs G2/M).
1. Registration process (One Time): New Registration for external users.User Verification: Upload photo of Front side and back side of valid Identity card issued by the organization (only .jpeg format, file Size less than 1 Mb, height less than 1000 pixels, width less than 800 pixels). Undertaking: Download the undertaking template ( Download Link ). Duly filled, signed and stamped form should be uploaded in .pdf format. Confirmation: An email will be sent to the user after verification of the documents.
2.Before planning to avail the Instrument facility of CRF IIT Delhi: user must visit https://crf.iitd.ac.in -> Facility -> select the desired facility from left side navigation list. Read carefully all the relevant information such as required sample type, testing charges and instrument location. Be prepared with details of the samples.
3.Log in and Click on Booking Appointment Tab: All the information relevant to testing must be filled in instrument booking form. In case user does not provide the required information the booking is liable to be rejected. User can upload the relevant existing data of their samples that might help in the measurements.
4.Sample submission & Payment Process: Once the application is submitted, the in-charge will verify the details and approve the request.External users must wait for the approval before making the payment and submitting the samples to concerned lab. After approval the payment should be made by the user. The payment transaction details need to be submitted online before appointment date time.
5.CRF Users can pay amount online using following details:
Beneficiary/Customer's Name: IRD ACCOUNT IITD
Bank Account Number: 10773572600
Bank Name: State Bank of India Branch Name: IIT Branch
Branch Code: 01077
NEFT IFSC CODE / RTGS: SBIN0001077
please write "CRF _Instrument Name" in the Remarks section while paying. Save the receipt file and upload in the crfbooking system online against your application.Payment can also made in form of demand draft in favour of " IRD ACCOUNT IITD " Payable at New Delhi.6.IIT Delhi
GST Number : 07AAATI0393L1ZI
PAN Number : AAATI0393L
1 Registration process (One Time): New Registration for internal users.
Fill in all the details in Personal Information, Supervisor Information and User Login Credentionals.
Once submitted, an email to supervisor is sent for activation of account. Once activated by supervisor, you can do the instrument bookings.
2 Before planning to avail the Instrument facility of CRF IIT Delhi: user must visit https://crf.iitd.ac.in > Facilities > select the desired facility from left side navigation list. Read ca 
refully all the relevant information such as required sample type, testing charges and instrument location. Be prepared with details of the samples.
3. Log in on CRF Booking System and Click on Booking Appointment Tab: All the information relevant to testing must be filled in instrument booking form. In case user does not provide the required information the booking is liable to be rejected. User can upload the relevant existing data of their samples that might help in the measurements.
4 Sample submission & Payment Process: Once the application is submitted, the in-charge will verify the details and approve the request.
